Understanding Wiring and Volume Transmission: Review

B RA I N RE SE A R CH RE V I EW S 64 ( 20 1 0 ) 1 3 71 5 9
available at www.sciencedirect.com
www.elsevier.com/locate/brainresrev
Review
Understanding wiring and volume transmission

Luigi F. Agnati a,, Diego Guidolin b , Michele Guescini c , Susanna Genedani a , Kjell Fuxe d
a
IRCCS San Camillo Venezia, Department of Biomedical Sciences, University of Modena and Reggio Emilia, Via Campi 287,
Modena 41100, Italy
b
Department of Human Anatomy and Physiology, University of Padova, Via Gabelli 65, Padova 35122, Italy
c
Department of Biomolecular Sciences, University of Urbino Carlo B, Via A. Saffi 2, Urbino 61029, Italy
d
Department of Neuroscience, Division of Cellular and Molecular Neurochemistry, Karolinska Institutet, Retzius vg 8,
Stockholm 17177, Sweden
A R T I C LE I N FO
AB S T R A C T
Article history:
The proposal on the existence of two main modes of intercellular communication in the
Accepted 17 March 2010
central nervous system (CNS) was introduced in 1986 and called wiring transmission (WT)
Available online 27 March 2010
and volume transmission (VT). The major criterion for this classification was the different
characteristics of the communication channel with physical boundaries well delimited in
Keywords:
the case of WT (axons and their synapses; gap junctions) but not in the case of VT (the
Wiring transmission
extracellular fluid filled tortuous channels of the extracellular space and the cerebrospinal
Volume transmission
fluid filled ventricular space and sub-arachnoidal space). The basic dichotomic classification
Tunnelling nanotube
of intercellular communication in the brain is still considered valid, but recent evidence on
Roamer type of VT
the existence of unsuspected specialized structures for intercellular communication, such
Microvescicle
as microvesicles (exosomes and shedding vesicles) and tunnelling nanotubes, calls for a
Communication network
refinement of the original classification model. The proposed updating is based on criteria
Central nervous system
which are deduced not only from these new findings but also from concepts offered by
informatics to classify the communication networks in the CNS. These criteria allowed the
identification also of new sub-classes of WT and VT, namely the tunnelling nanotube type
of WT and the Roamer type of VT. In this novel type of VT microvesicles are safe vesicular
carriers for targeted intercellular communication of proteins, mtDNA and RNA in the CNS
flowing in the extracellular fluid along energy gradients to reach target cells. In the
tunnelling nanotubes proteins, mtDNA and RNA can migrate as well as entire organelles
such as mitochondria. Although the existence and the role of these new types of
intercellular communication in the CNS are still a matter of investigation and remain to
be fully demonstrated, the potential importance of these novel types of WT and VT for brain
function in health and disease is discussed.
2010 Elsevier B.V. All rights reserved.
Corresponding author. IRCCS San Camillo via Alberoni 70, Lido VE, Italy.
E-mail address: luigiagnati@tin.it (L.F. Agnati).
Abbreviations: AD, Alzheimer's disease; ASIC1a, acid-sensing ion channel 1a; CCNs, complex cellular networks; CL, cytoplasmic loop;
CNS, central nervous system; CSF, cerebrospinal fluid; Cxs, connexins; ECF, extracellular fluid; ECM, extracellular matrix; ECS,
extracellular space; GJ, gap junction; GMN, global molecular network; ILVs, intra-lumenal vesicles; MVB, multivesicular bodies; TM,
transmembrane domain; TNTs, tunnelling nanotubes; UCP, uncoupling protein; VT, volume transmission; WT, wiring transmission
0165-0173/$ see front matter 2010 Elsevier B.V. All rights reserved.
doi:10.1016/j.brainresrev.2010.03.003
138
B RA I N R E SE A R CH RE V I EW S 64 ( 20 1 0 ) 1 3 71 5 9
Contents
1.
Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
1.1. Wiring transmission and volume transmission . . . . . . . . . . . . . . . . . . . . . . . . . . . .
2.
Wiring transmission . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
2.1. Classical chemical synaptic transmission . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
2.2. Gap junctions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
2.3. Mixed synapses . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
3.
Volume transmission . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
3.1. Ephaptic transmission or electrical VT. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
3.2. Perisynaptic VT . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
3.3. Classical volume transmission beyond the perisynaptic region . . . . . . . . . . . . . . . . . . .
3.4. Possible interactions between transmitter spill-over and ephaptic effects at chemical synapses .
4.
Putative novel types of wiring and volume transmission . . . . . . . . . . . . . . . . . . . . . . . . . .
4.1. Tunnelling nanotubes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
4.2. Roamer type of VT. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
5.
On the role of the extracellular space in intercellular communication . . . . . . . . . . . . . . . . . . .
6.
Concluding remarks . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
1.
Introduction
In 1986 we published our original proposal on the existence

of two main modes of intercellular communication in the
CNS: the wiring transmission (WT) and the volume transmission (VT) (Agnati et al., 1986), which was aimed to
complement the Cajal's and Sherrington's view of the central
nervous system (CNS) as a computational apparatus basically formed by neurons interacting via specialized sites of
contiguity, namely the synaptic contacts (Cajal, 1906;
Sherrington, 1906) (Fig. 1). Our proposal was influenced by
previous important contributions on communication in the
CNS (Golgi, 1914; Guillemin, 1978; Nicholson, 1979; Schmitt,
1984; Descarries et al., 1991; Nieuwenhuys, 2000; Bach-Y-Rita,
2005) and based on a number of observations, especially on
the central monoamine neurons (for reviews: Fuxe and
Agnati, 1991a,b; Agnati and Fuxe, 2000). The concept of VT
introduced the extracellular space and the ventricular
system as important channels for chemical transmission in
the CNS complementary to WT with diffusion and flow of
transmitters, ions, trophic factors, ... in the extracellular fluid
(ECF) and cerebrospinal fluid (CSF). Furthermore, our proposal in 1986 considered that the VT signals could interconnect not only neurons in neuronal networks but rather cells
of any type (neurons, glia, microglia, ependymal cells,
macrophages,) in what we have called the complex
cellular networks (CCNs) of the brain (Agnati and Fuxe,
2000). Thus, these signals migrating in the ECF of the CNS
could affect multiple targets in the CCNs.
In some instances they could even lead to a new output
from the same CCN as surmised within the concept of
polymorphic networks (Getting and Denkin, 1985) (see
Fig. 2 for an example of a VT modulation of a polymorphic
neuronal network). The existence of VT has also made it
possible to suggest different informational models for memory processes (Agnati and Fuxe, 2000; Guidolin et al., 2007) and
its relevance for neuropsychopharmacology (Zoli et al., 1999)
has been underlined.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
138
138
139
140
140
142
142
145
145
146
146
147
147
149
151
152
154
It is our opinion that the basic dichotomic classification of

intercellular communication in the brain proposed more
than two decades ago is still valid. However, we are fully
aware that evidence on the existence of new specialized
structures for intercellular communication, such as microvesicles (for a review, see Cocucci et al., 2009) and tunnelling
nanotubes (Rustom et al., 2004; Baluka et al., 2006; Goncharova and Tarakanov, 2008) (Fig. 3), calls for an updating of our
original conceptual model. This also includes the introduction by our concept that communication via both WT and VT
belongs to the fundamental features of all neurons, the ratio
of which can vary from one neuron system to the other and
with the structural and functional state of each neuron
system.
The criteria which can be applied to characterize WT and
VT and their sub-classes will be deduced not only from
structural and neurochemical findings, but also from concepts
and from the lexicon offered by informatics (Hopcroft and
Ullman, 1979). Our presentation will follow a historical frame
moving from the well established modes toward novel modes
of cellcell communications. Thus, after a condensed summary of the main features of the classical modes of WT and
VT (Agnati and Fuxe, 2000) tunnelling nanotubes and microvesicles will be discussed as novel types of WT and VT,
respectively.
Furthermore, recent experimental findings of our group on
these new types of WT and VT will be presented and, finally,
their potential relevance for the physiology and pathology of
the CNS will be discussed.
1.1.
Wiring transmission and volume transmission
Taking advantage of some concepts and the lexicon offered by

informatics (Hopcroft and Ullman, 1979; Le Boudec and
Thiran, 2001), it becomes possible to classify the intercellular
communication in the CNS according to a series of criteria
based on the characteristics of the communication channel, of
the transmitted signal and of the formed network. They are
139
Fig. 1 According to Kuhn's epistemology a paradigm allows to evaluate the relevance of facts in a given science (Kuhn, 1962).
However, any paradigm has a physiological life span, since it is sooner or later replaced by a new paradigm, which explains
experimental evidence that could not be explained by the old paradigm. Thus, it occurs what Kuhn has called an
epistemological paradigm shift. The scheme in the figure illustrates the classical paradigm of inter-neuronal
communication in the brain on which most of the approaches to Neurophysiology and Neuropathology have been founded. As
indicated in the figure, this conceptual model was proposed by Cajal and Sherrington at the beginning of the last Century
(Sherrington, 1906). For further details, see Agnati et al. (2007c).
illustrated in Table 1, where a possible approach (Ramanathan

et al., 2007) to a more formal definition is also outlined.
In this respect it has to be observed that the one and main
criterion which allows differentiating WT from VT are the
characteristics of the communication channel and more
precisely the physical boundaries of the channel, which are
well delimited for WT but not for VT.
The classification (in particular when VT based intercellular communication pathways are concerned), however, could
be further detailed by taking into account the other signal
features illustrated in Table 1 (see also legend to Table 2):
Signal privacy: It is proposed that we are dealing with a signal
characterized by high privacy (i.e., a reserved signal) if only
cells endowed with a specific recognition/decoding apparatus (as, for instance, specific receptors) can have access to it.
On the contrary, we are dealing with a low privacy signal
(i.e., a broadcasted signal) when any cell reached by the
signal can have access to it.
Signal safety: As far as the safety is concerned, we are dealing
with a safe signal if it is not altered during its conduction
from the source to the target cell and with an unsafe signal
if it can be altered during its pathway, as occurring, for

instance, to some VT signals that can be broken down or
modified (e.g., by enzymes) in the extracellular space (ECS).
Connectivity: If the connections between cells can be rapidly
formed or removed, they provide a dynamic network. On
the contrary, the structure of a communication network is
static when the pattern of connections is almost stable in
time.
Thus, based on these concepts, a unitary scheme could be
devised for a more detailed characterization of WT and VT and
of their subtypes. Let us now briefly examine them.
2.
Wiring transmission
The specific feature of this mode of intercellular communication

is the existence of a virtual wire connecting the cell source of the
signal (message) with the cell target of the signal. The different
subclasses of WT are indicated in Table 2, together with the
properties they exhibit in terms of the criteria to classify modes
of communication in cellular networks of the CNS (Table 1).
140
Fig. 2 Changes in the mix of VT and WT can also give rise to polymorphic networks that is to a neural network, which can give
different outputs according to the VT signals impinging on it. An example of a polymorphic network is illustrated for a neuronal
network where the VT signals by up-regulating or down-regulating synaptic contacts can change the integrative action of the
network. Thus, as shown in this schematic drawing VT signals via modulatory actions on the synaptic contacts of the neuronal
network can give rise to three types of outputs (Panel A: Output 1, Panel B: Output 2, Panel C: Output 3). Obviously, the same
conceptual scheme can be applied to a Complex Cellular Network.
The most important and well known is certainly the

synaptic transmission. However, two more subclasses of WT
could play some role in the CNS. The first one is represented
by the well characterized gap junctions, while the second one,
the clear-cut in vivo demonstration of which has not yet been
provided, is represented by the tunnelling nanotubes (TNTs)
(Rustom et al., 2004). The WT mode of intercellular communication is characterized by a channel representing, at least
transiently, a truly continuous wire connecting the source of
the message with the target of the message and therefore all
the above mentioned forms of WT share a high level of signal
safety. Among them, however, only the synaptic transmission
is also characterized by a high level of signal privacy based on
specific receptor systems to decode the transmitted signal
(Table 2). Since TNTs represent a quite novel finding, they will
be described in more detail than the other types of WT (see
Section 4.1).
2.1.
Classical chemical synaptic transmission
It is beyond question that classic synaptic wiring, which

depends on the transmission of action potentials, is the most
important example of WT and the primary mechanism
through which neurons transmit information and control
behavior. It represents the prototype of the WT since it is

characterized by a virtually continuous wire connecting the
source of the message with its targets. Actually, a small gap
(the synaptic cleft less than 50 nm) in the channel is present
between the site of release of the transmitter and the site
where the receptors capable of detecting and transducing the
signal are located. However, in the classical synaptic transmission it is assumed that this gap does not interfere with the
continuity of the channel (Savtchenko and Rusakov, 2007) and
with the safety of the message. Moreover, as mentioned
before, the signal can be classified as reserved since specific
receptors are needed to decode the message (Table 2). This WT
will be further discussed in relation to perisynaptic VT
transmission.
2.2.
Gap junctions
Connexins (Cxs), a large family of homologous membrane

proteins in vertebrates, form gap junction (GJ) channels that
provide a direct pathway for electrical and metabolic signaling
between cells, especially astrocytes (LeBeau et al., 2003; Herv
et al., 2007; Yeager and Harris, 2007). Each GJ channel, which
has an inner pore diameter of 1012 Angs, is composed of two
hemi-channels (connexons), which in turn are composed of
141
Fig. 3 Schematic representation of the main features of tunnelling nanotubes and microvesicles. According to several
authors (see, e.g., Cocucci et al., 2009 and Table 3) Microvesicles can be subdivided into two main classes, exosomes and
shedding vesicles. Both types of Microvesicles play a role as safe vesicular carriers for targeted intercellular communication
via VT allowing the transfer of packets of signals to target cells. Exosomes are microvesicles with a diameter of 40100 nm
that originate in multivesicular-bodies of the endosomal system. Exosomes via exocytosis are released both constitutively
and in a regulated manner. It is also illustrated that the exosomes can undergo endocytosis as they impinge on the target
cell. Other endosomes are shown to be inserted into the plasma membrane and into organelles. Shedding vesicles are
formed from lipid raft domains of the plasma membrane. It should be mentioned that lipid rafts often contain signalosomes,
which are organized clusters of macromolecules involved in the recognition/decoding of chemical signals, including the
receptor mosaics. It is illustrated how they may be formed by budding from lipid rafts and also how they may impinge on the
plasma membrane of target cells and may transfer lipid rafts with associated signalosomes. For further details, see Table 4
and text.
six Cx subunits. Connexons can be coupled with a hemichannel of an adjacent cell to form a gap junction. Each of the
six Cx subunits forming a connexon has four alpha helical
transmembrane domains (TM1 to TM4), intracellular N- and Ctermini, two extracellular loops, and a cytoplasmic loop (CL).
Since at least 20 distinct Cx isoforms have been cloned, the
question has been raised as to their possible different
functional meaning. Actually, channels vary in permeability
selectivity from being non-selective, or being preferentially
selective for cations or anions (Oviedo-Orta and Evans, 2004).
Gap junctions are also found between activated microglia,
between many types of neurons, between astrocytes and
oligodendrocytes (Orthmann-Murphy et al., 2008), and in a few
somewhat controversial instances between astrocytes and
neurons (Nagya et al., 2004).
Gap junctions are evenly distributed along the astrocytic
processes, often interconnecting adjacent astrocytic processes
derived from the same cell. However, usually different astro-
cytes are coupled through gap junctions to form large intercellular networks (Theis et al., 2005; Volterra and Meldolesi, 2005).
The function of these gap junctions is to minimize the
differences not only between individual processes of the same
astrocyte but also between astrocytes by mediating the sharing
of substrates such as glucose. Furthermore, they play a role in
dissipating extracellular K+ or glutamate, whose extracellular
accumulation can be detrimental for the proper neuronal
functions. Thus, the communication network generated by
gap junctions in astrocytes is also of significant support to the
main processing network formed by the synaptic wiring among
neurons. The opening probability of the hemi-channels
increases markedly by reducing the concentrations of divalent
cations, notably Ca2+, and is accompanied by the release of both
ATP and glutamate. The control of opening probability is
important, since gap junction channels allow the coordination
of intrinsic or elicited metabolic and/or electrical responses of
cells in a heterogeneous population. Channel conductance and
142
Table 1 Possible informatics criteria to classify communication networks of CNS.

Network feature
Connectivity
Types
Static
Dynamic
Signal privacy
Reserved
Broadcast
Signal safety
Channel (Main criterion to distinguish

volume transmission from
wiring transmission)
Unsafe
Safe
Private
Diffuse
Description
The network structure does not change with time.
From a formal point of view it can be defined as:
G = (V,E)
where G is a graph, i.e., a 2-ple composed by
V = { vi, 1 i N} the set of vertices (nodes)
E = { (u,v): u,v V} the set of edges (connections)
The network has a time-varying structure
At a particular instant of time t it can be described as:
G(t) = (V(t), E(t))
Where:
E(t) = {(u,v): u,vV(t) and communicating at time t}
Signal needing a specific decoder to be decrypted
Neurotransmitters and, more generally, signals using specific receptor
systems are of this type
Public signal, i.e., interpreted by all the involved elements
Physical quantities or membrane permeable molecules are of this type
The signal can be altered during its travel from the source node to the
destination node
The signal comes to the destination node without alteration
Physically delimited pathway between two nodes of the network.
This type of channel characterizes WT
The whole available space between the network nodes is

potentially used to exchange signals.
Thus, no end-to-end path is present between two
communicating nodes, but there is a temporally ordered
sequence of events that connects the two nodes.
This type of channel characterizes VT
permeability are under a control carried out by changes in the

gating mechanisms and by changes in the Cx turnover (Goldberg
et al., 2002). The half-life of several rodent connexins has been
found to be between 2 and 5 h (for reviews: Saez et al., 2003a,b;
Contreras et al., 2004; Bloomfield and Vlgyi, 2009).
2.3.
Mixed synapses
Gap junctions can also be observed in chemical synapses,

hence mixed synapses are possible, which may have a
significant modulatory role on the neural networks, imparting
fundamental alterations to their properties. Thus, mixed
synapses provide the structural and functional components
required for both chemical and electrical transmission within
a single synaptic contact on an individual postsynaptic
element (Rash et al., 1996; Flores et al., 2008). The presence
of both types of synaptic communications allows a more
subtle and complex neuronal computation than might occur
with only one type of synaptic contact. It has been surmised
that in the mixed synapse direct physical interactions
between receptors and connexins could play an important
role (see Fuxe et al., 2007a). It should be considered that also
some additional features are added to the exchange of signals
from the pre-synaptic to the post-synaptic side thanks to the
presence of a gap junction, since the transient interconnec-
tions of the two cytoplasms via the gap junctions allow the
protected intercellular transfer of second messengers, such as
IP3 and Ca2+ and other low molecular weight substances such
as small peptides (less than 1000 Da (Oviedo-Orta and Evans,
2004)), and also of siRNAs (Valiunas et al., 2005). Furthermore,
it should also be mentioned that some gap junctions (nonrectifying gap junctions) allow the bidirectional passage of the
electrical signal (Nagya et al., 2004; Gonzlez et al., 2007). It has
to be pointed out, however, that in the mammalian CNS, the
physiological roles and distributions of mixed synapses are
still to be clearly established (Rash et al., 1996).
3.
Volume transmission
Interneuronal wiring communication is certainly a basic

feature of the CNS and has been the main foundation of
neuroscience as we know it. In the 1970s and 1980s, however,
the functional assumption of a diffuse mode of intercellular
communication affecting and modulating the activity of entire
brain regions was gaining support from studies on monoaminergic (see Ungerstedt et al., 1969; Geffen et al., 1976) and
peptidergic neurons (Fuxe et al., 1977; Bloom and Segal, 1980;
Burbach, 1982; De Wied and Jolles, 1982) and led to the
definition of VT (Agnati et al., 1986).
143
Table 2 Summary of the different types of wiring transmission and volume transmission forming the communication
networks of the brain ( see Table 1).
Network
Synaptic transmission (Section 1.1)
Gap junctions (Section 1.2)
Ephaptic transmission (Section 2.1)
Perisynaptic transmission (Section 2.2)
Classical volume transmission (beyond
the perisynaptic region) (Section 2.3)
TNT (Section 3.1)
Roamer type of VT (Section 23.2)
Signal privacy
Signal safety
Channel
Connectivity
Reserved
Broadcast
Broadcast
Reserved
Reserved (common)
or broadcast (rare)
Broadcast
Reserved or Broadcast
Safe
Safe
Safe
Unsafe
Unsafe
Private (WT)
Private (WT)
Diffuse (VT)
Diffuse (VT)
Diffuse (VT)
Static and/or dynamic a

Dynamic
Dynamic b
Dynamic
Dynamic
Safe
Safe
Private (WT)
Diffuse (VT)
Dynamic
Dynamic
Definitions of the terms:
Reserved: a specialized receptor apparatus is needed to decode the message

Broadcast: no specialized receptor apparatus is needed to decode the message
Private: the channel transmit signals along a conductor with clear cut physical boundaries
Diffuse: the channel transmits signals along a conductor without clear cut physical boundaries, hence signals can
potentially migrate in any direction
- Safe: the signal is not altered during its conduction from the source to the target
- Static: the connection between the source and the target is maintained for relatively long periods of time (sometimes for
a life-long period)
- Dynamic: the connection between the source and the target can be rapidly formed and/or removed.
a
Synaptic transmission is endowed with a potential high plasticity; hence it could give rise to neural networks characterized by different types
of connectivity. They include networks that once established exhibit an almost stable (static) configuration (as, for instance in the primary visual
cortex; Hensch, 2005), as well as dynamic networks in which a number of connections are ex novo established (see, for instance, the neural
circuitry responsible for seasonal breading in several species; Adams et al., 2006) or re-activated (see Kerchner and Nicoll, 2008 for a review on
silent synapses) whenever needed.
b
Ephaptic transmission is highly dependent on the volume fraction of the ECS and the composition of the ECM, which can affect the diffusion
pathways of ion currents (see also the text).
VT is characterized by the absence of any wire-like channel

connecting the source of the signal with its own targets and
this feature usually leads to reduced safety since the VT signal
migrates in the ECF (Tables 1 and 2). This communication
mode uses several often spatially divergent tortuous channels
made by the clefts (about 20 nm in diameters; Chen and
Nicholson, 2000) between cells and filled with extracellular
fluid and extracellular matrix. Signals migrate in the extracellular space and may be stopped if they reach a blind alley (a
cul-de-sac) (Hrabetov et al., 2003), if they are inactivated by
enzymes or cleared over the brain capillaries (Jansson, 2000) or
taken up into cells via transporters (Rice and Cragg, 2008). VT
signals can be released from any type of brain cells, hence
from dendrites, soma and axon terminals (varicosities)
entirely lacking synaptic membrane specializations (asynaptic) (Descarries et al., 2008), or endowed with such specializations, and from astroglial, microglial, oligodendroglial and
ependymal cells.
As detailed in Table 3, where the available supporting
evidence is reported, VT can employ the same set of signals as
WT, namely transmitters and ions. Other types of signals,
however, were suggested to be linked to VT (see Getting and
Denkin, 1985; Agnati et al., 1994, 1995a,b, 2005a; Fuxe et al.,
2010). Thus, it is possible to list the VT signals as follows:
- Classical and non-classical intercellular chemical mediators: neurotransmitters, neuromodulators, growth factors,
ions (e.g., Ca2+), gases (NO, CO2, CO);
- Physical signals (as, for instance, field potentials).
Moreover, it is important to distinguish lipophilic from

hydrophilic VT signals, since while the first ones can diffuse
also across membranes (i.e., they have a large space of
diffusion) the latter ones are practically confined to the ECS.
Let us briefly examine some of these VT signals and their
main features. As stated in Table 2 most of the VT signals are
reserved, rather than broadcasted, and unsafe. However, we
should mention two typical examples of broadcasted VT
signals, the field potentials and the CO2, which can be also
related to each other via their effects on the membrane
potential of neurons and astrocytes. As a matter of fact,
evidence has been provided that carbonic anhydrase-II is
localized in the CNS to wide spread systems of oligodendrocytes and restricted astroglia populations, involving both fiber
bundles and neuropil (Agnati et al., 1995d). It has also been
suggested that CO2 formed in activated axons may, via
carbonic anhydrase-II, give rise to protons controlling the
excitability of surrounding neuropil (see Sun and Alkon, 2002).
Thus, CO2 may represent an important, highly diffusible,
broadcasted" signal in classical VT in the brain, involved via
its modulatory action on pH in the tonic control of neuronal
excitability and in protein 3D-conformations (see Petsko and
Ringe, 2004).
It is also important to note that CO2 can also operate as a
reserved signal, since the acid-sensing ion channel 1a
(ASIC1a), which is localized in dendritic spines, functions as
a proton receptor. Such a channel can modulate the local
circuitry of the neuronal network, because it affects the
density of spines, which are the postsynaptic site of most
144
Table 3 Chemical mediators involved in VT.

Signaling
molecule
5-HT
Glutamate
DA
Animal
Rat
Rat
Rat
Mouse
Brain regions
Hippocampus
Frontal cortex
Cerebellum
Dorsal raphe nucleus
or Substantia nigra
pars reticulata
Spinal cord: dorsal and
ventral horn
Hippocampal slices
Cerebellar slices
Ventral pallidum
Ventral subiculum
Pedunculopontine
nucleus
Findings
Existence of extrasynaptic receptors and
transporters for 5-HT
Diffusion of 5-HT for a few microns
5-HT receptors on neurons not receiving a
direct 5-HT innervation
5-HT fibers often lack typical synaptic contacts
Electrochemical studies show that 5-HT can
enter the extracellular space
5-HT diffusion in the order of 8 m
Glutamate spill-over
Astroglia processes and neuronal glutamate
transporters modulate spill-over
Astroglial release of glutamate
Synaptically released glutamate can exert
long-range actions in the cortical microcircuitry
NMDARs involved in the response to
extrasynaptically released glutamate
Extrasynaptic tonic DA release
Diffusion is the most important determinant
of DA time-course
VT model of DA varicosity
NE
Mouse
Brain slices
Prefrontal cortex
Brain slices
Hippocampus
Neocortex
Brainstem slices
containing the superior
olivary complex
Mid brain
Cerebral cortex
Ca2+
Rat
NO
Mouse
-endorphin
Rat
Galanin
Rat
Ach
Rat
GABA
Rat
Raphe region
Amygdale
Parietal cortex
Hippocampus
Somatosensory cortex
CO2
Rat
Hippocampus
Mismatch between D4 IR , TH IR and

-hydroxylase IR
NE remains in the extracellular space for a time
sufficient to travel 100 m
Presence of calcium sensing G protein-coupled
receptors in the brain
Suppresses postsynaptic Kv3 potassium currents
Inhibits postsynaptic glutamate receptors, without
changing transmitter release
Released -endorphin after stimulation of arcuate
nucleus reaches other brain areas and affect brain
function in a global manner
Inhibitory modulation of 5- HT1A receptor affinity
and efficacy.
Faster form of non-synaptic transmission
operating at short distances
GABA released from neurogliaform cells acts as a
volume transmitter
Alterations in carbonic anhydrase activity in
hippocampal CA1 neurons provide a mechanism
for switching between operational states at
GABA-releasing synapses, thereby gating signal
transfer through the hippocampal network
excitatory synapses. Thus, decreasing ASIC1a reduces the

number of spines, whereas over-expressing ASIC1a has the
opposite effect (Zha et al., 2006).
In this brief list of VT signals, two highly lipophilic
reserved signals, namely the gases NO and CO, which are
major neurotransmitters (Baraano et al., 2001) should be
mentioned. Their main molecular target is an intracellular
enzyme (the heme moiety of soluble guanylyl cyclase), which
is not a proper receptor but gives specificity to their action. In
addition CO formed from heme oxygenase 2 specifically binds
to the transcription factor NPAS2, participating in circadian
References
Bunin and Wightman (1999)
Ridet and Privat (2000)
Doly et al. (2004)
Ciranna (2006)
Bunin and Wightman (1998)

Jansson et al. (1998, 2001, 2002)
Descarries & Mechawar (2000)
Kullmann et al. (1996)
Del Arco et al. (2003)
Szapiro & Barbour (2007)
Piet et al. (2004)
Diamond (2001)
Scimemi et al. (2004)
Clark & Cull-Candy (2002)
Floresco et al. (2003)
Goto et al. (2007)
Floresco (2007)
Cragg et al. (2001)
Rice & Cragg (2008)
Rivera et al. (2006)
Rivera et al. (2008)
Fuxe et al. (2003)
Mundorf et al. (2001)
Yano et al. (2004)
Steinert et al. (2008)
MacMillan et al. (1998)
Fuxe et al. (1998)

Lendvai & Vizi (2008)
Descarries & Mechawar (2000)
Olh et al. (2009)
Sun & Alkon (2002)
rhythms and regulating its DNA binding activity (Boehning

and Snyder, 2002).
It has been shown that also high-molecular weight
proteins (e.g., prolactin and interleukin1-beta) can participate
in classical VT, migrating in the extracellular fluid of the brain
and signaling to target cells (Fuxe et al., 1977; Bjelke et al., 1988;
Jansson, 2000; Fuxe et al., 2007a) and this is also true for large
peptides such as Abeta (Agnati et al., 2007b; Leo et al., 2007).
Furthermore, it has also been proposed that classical VT
communication is involved in intercellular RNA transfer, since
RNAs may be secreted into extracellular spaces despite their
susceptibility to degradation by extracellular RNAses (Alvarez

et al., 2000; Eyman et al., 2007; Smalheiser, 2007; Bicker and
Schratt, 2008); indeed, extracellular RNAses appear to have a
regulatory role in cell physiology, which may be a clue to
indicate that the secreted RNAs comprise a VT form of cellcell
communication.
Based on the criteria reported in Table 1, different
subclasses of VT could be differentiated. They are summarized
in Table 2 and discussed in the next sections.
3.1.
Ephaptic transmission or electrical VT
This was the first type of VT to be proposed to exist in the

CNS (Golgi, 1891) and the main one to inspire us in proposing
the VT mode as a significant communication mode in the
brain in view of its self-evident existence deriving from
physical concepts already well established by Volta two
centuries ago (Trasatti, 1999). As a matter of fact, Golgi
postulated it by stating that the material contact between
neurons is not a necessary condition for their communication
by electrical signals. As indicated above, Golgi based his bold
statement on Volta's studies on the second class of conductors, i.e., the so-called volume conductors, which are
electrolytic solutions, such as body fluids. Thus, ephaptic
transmission involves electrical signaling due to the flow of
ions between neurons and/or axons through a volume
conductor, the ECF. In agreement with Golgi's proposal,
changes in excitability evoked by passive current flows
between adjacent cells were described by Arvanitaki (1942)
as opposed to synaptic transmissions at sites of specialized
junctions and Kamermans and Fahrenfort (2004) provided
evidence of ephaptic modulation within a chemical synapse.
It should be noted that even if the term ephaptic comes from
the Greek word (ephaptein) which means to bind,
ephaptic interactions occur between neurons or parts of
neurons in close contiguity but not in continuity with each
other (Fig. 1).
It has been observed that ephaptic interactions between a
neuron and axons or dendrites passing by its cell body can be,
in principle, more significant than ephaptic interactions
among axons in a fiber tract. Extracellular action potentials
outside axons are small in amplitude and spatially spread out,
while they are larger in amplitude and much more spatially
confined near cell bodies (Holt and Koch, 1999). Depending on
the direction of the predominant current flow near the sites of
spike initiation, mainly excitatory or inhibitory effects could
be predicted theoretically and demonstrated in experiments
(Krnjevic, 1986).
The ionic currents of ephaptic transmission can generally
affect all the cells they reach independent of receptors and
therefore act as broadcasted signals. A special case is
represented by the calcium ion fluxes since there does exist
a calcium sensing G protein-coupled receptor (Brown et al.,
1993) also in the brain especially in the glial networks. Calcium
ion fluxes should, therefore, be considered also as a signal of
the reserved type (Yano et al., 2004). The safety of ephaptic
transmission is high since the volume fraction of the
extracellular space allows substantial ion fluxes to take place
(Nicholson, 2000; Sykov et al., 2000). However, the shrinkage
of the volume fraction via cell swelling may increase the brain
145
tissue electrical resistance and reduce ephaptic transmission

as well as VT in general (Sykov et al., 2000).
3.2.
Perisynaptic VT
Perisynaptic VT is linked to synaptic transmission and likely

takes place to a varying degree as a consequence of synaptic
transmission due to incomplete diffusion barriers, with the
synaptic transmitter reaching the perisynaptic domains of the
pre- and post-synaptic membrane, the astroglia and even
adjacent synapses. Thus, glial sheaths around the synapse
and the extracellular matrix (ECM), forming perineuronal nets
(Celio et al., 1998) act as diffusion barriers. In addition, the fate
of the escaping transmitter molecules is determined by the
activity of the transmitter transporters and/or the inactivating
enzymes. Thus, the glial envelope of the synapse and its
membrane specializations are the hallmark of synaptic
transmission favoring a point to point communication.
Nevertheless, even in the classical synapse where signaling occurs through the private synaptic cleft, the insulation
of the synapses is often insufficient to prevent perisynaptic
VT (Oliet et al., 2006; Witcher et al., 2010) with the diffusion of
mediators from the synaptic cleft reaching perisynaptic
regions and astroglia rich in receptors (Deitmer and Rose,
2010; Cabello et al., 2009) and finally leading to synaptic spillover to other synapses (Piet et al., 2004; Alle and Geiger,
2007). However, what is the relative importance of this type of
transmission? It seems that both synaptic transmission and
perisynaptic VT belong to the fundamental properties of
neurons and both participate in the integrative processes of
the local circuits (Schmitt, 1984; Ferr et al., 2007; see Agnati
et al., 2007a for a review). Thus, during repetitive stimulation,
when the pre-synaptic terminals of two neighboring synapses
are near to one another and a paucity of glial membranes is
interposed, synaptic spill-over favors their crosstalk. As a
matter of fact, it is accepted that via diffusion, the classical
neurotransmitters glutamate or GABA can reach receptors at
astroglia and at neighboring synapses affecting the efficacy of
transmission (Reichenbach et al., 2010), a process regulated
also by the affinity of the glial and neuronal transporters of
glutamate and GABA and also leading to astroglial release of
glutamate (Olh et al., 2007; see Fuxe et al., 2007c, 2010). This
important field of investigation was initiated in the midnineties mainly by Gonon and Kullmann, who proposed
neurotransmitter spill-over as one mechanism for intersynaptic crosstalk (Gonon et al., 2000; Kullmann, 2000).
Glutamate released during synaptic events could escape the
synaptic cleft and reach NMDA receptors at neighboring
synapses (Kullmann et al., 1996; Asztely et al., 1997). Thus,
when glutamate escapes from a synaptic cleft or is released
by astrocytes, it reaches a relatively high concentration in
narrow extracellular clefts and diffuses from the place of
release if it is able to overcome not only the physical barriers
to diffusion (Nicholson and Sykova, 1998; Sykov and
Vargov, 2008) but also the buffering action of the uptake
systems present in neurons and astrocytes (Rice and Cragg,
2008). It has been suggested that this type of intercellular
communication plays a role in long-term potentiation and
depression, during lactation or dehydration, where it can
potentiate neurohormonal release (Oliet et al., 2001; Sykova,
146
2001; Piet et al., 2004), or in the indirect modulation of the

dopaminergic system by excitatory inputs (Kiss et al., 2004). It
should be noted that this type of cellcell communication is
plastic, since the astrocytic isolation of synapses may change
over either a short time-scale (e.g., during neuronal activity;
Hirrlinger et al., 2004) or over a long time-scale (e.g., during
lactation (Theodosis and Poulain, 1993; Haydon, 2001; Sykova,
2001)).
It appears that the activation of extra-synaptic receptors at
and around the postsynaptic target may be tightly regulated.
In contrast, in some instances, glutamate is allowed to diffuse
relatively freely to receptors on and around pre-synaptic
terminals (Barbour, 2001). This appears to allow the same
neurotransmitter to carry two types of information: a highly
focused orthodromic signal that can be modulated by the
action of neuronal and glial transporters, and a more
widespread and diffuse signal to presynaptic and heterosynaptic structures, detected by metabotropic, kainate and
possibly NMDA receptors (Rusakov and Lehreb, 2002). It is
clear that one and the same transmitter released by one and
the same neuron can operate both as a synaptic and as a
perisynaptic signal.
Also GABA can operate as a perisynaptic transmitter in
several brain regions and extra-synaptic GABAA receptors
have been shown to have a higher affinity to GABA and slower
kinetics of desensitization than the corresponding synaptic
GABAA receptors (Kullmann et al., 2005). This allows them to
mediate persistent tonic current, which is determined by
several factors such as the density of extra-synaptic GABAA
receptors, the proximity of these receptors to the GABA release
sources, and the density of GABA transporters near the GABA
release sources. Instead the GABAB receptors are mainly
located extra-synaptically (Charles et al., 2001) suggesting a
major role in perisynaptic GABA VT.
It should also be considered that VT signals likely can
diffuse into an unshielded synaptic cleft and if receptors
capable of detecting these VT signals are present, complex
WT/VT integrative interactions can occur at both pre- and
post-synaptic level. This can be the case for DA which mainly
operates as a VT signal (Fuxe et al., 2007c). Thus, D2-NMDA
heteromers may exist in cortico-striatal glutamate synapses
(Liu et al., 2006) which when reached by diffusing DA can lead
to integration of WT and VT transmission through tuning of
the NMDA mediated glutamate signaling in these heteromers
via D2-NMDA receptorreceptor interactions (Fuxe et al., 2008).
3.3.
Classical volume transmission beyond the
perisynaptic region
This type of VT is characterized by longer distances of VT
migration (Agnati et al., 1986; Fuxe and Agnati, 1991a,b) and is
probably the major mode of communication in monoamine
and peptide neurons in the CNS (Fuxe and Agnati, 1991a,b;
Agnati and Fuxe, 1996, 2000; Fuxe et al., 2007c, 2010). This is
evident by the frequent existence of non-junctional varicosities (Descarries and Mechawar, 2000; Descarries et al., 2008),
and a dominance of extra-synaptic receptors and transmitterreceptor mismatches in these neurons (Agnati et al. 1986;
Agnati and Fuxe, 1996; Jansson et al., 2001, 2002; Fuxe et al.,
2007a, 2010). The classical VT beyond the perisynaptic region
usually exhibits a low signal safety (Table 2 and Agnati et al.,

2006a; Fuxe et al., 2007a, 2010) and is characterized by signals
of the reserved type, since the VT signal is decoded by specific
receptors, with the exception of CO2 (for more details see
Section 3).
The signals (released by any type of cells) involved in this
type of VT are driven toward the target cells (any type of cells)
by several energy gradients resulting not only in diffusion but
also in flow of the VT signal in moving ECF (vector mediated
migration). The energy gradients for VT signal migration are:
(a) concentration gradients of the VT signal (diffusion); (b)
gradients of electrical potentials (for charged signals); (c)
pressure gradients (vector mediated migration) (see the tide
hypothesis Agnati et al., 2005a); (d) temperature macrogradients (i.e., between a brain active region and a brain
inactive region (Yablonskiy et al., 2000) and micro-gradients
via uncoupling protein (UCP) (vector mediated migration)
(Agnati et al., 2005a,b; Fuxe et al., 2005; Rivera et al., 2006). It
should be noted that pressure and temperature gradients
besides enabling the extracellular fluid renewal (i.e., the
homeostasis of the brain internal milieu), also favor VT signal
migration. Thus, VT occurs both at lower costs in terms of
energy than the classical synaptic transmission (the prototype
of WT), and with much lower space filling since VT does not
need a dedicated channel but takes advantage of the ECS
(Agnati et al., 2005a, 2007c).
Of particular importance is the role played by the CSF,
which flows through ventricles, subarachnoidal spaces, and
the porous parenchyma in a pulsatile manner (Tawse, 2008).
As demonstrated by Greitz (1993, 2006), pressure waves
generated by the rhythmic cardiac activity pervade and
deform the entire mass of the brain. Such a cyclic phenomenon has profound effects on the dynamics of cerebral blood
and CSF flow and hence on the migration of signals in the
extracellular space of the brain (see the Tide Hypothesis
Agnati et al., 2005a, 2006a). Pressure waves could also exert
effects on the stretch sensitive ion channels present in some
neurons and astrocytes (Puro, 1991; Ostrow and Sachs, 2005;
Honor, 2007) and also on the NMDA channels, which respond
to membrane tension (Paoletti and Ascher, 1994).
Very recently this physical phenomenon has been carefully
evaluated by means of MRI and computer-assisted image
analysis (Linninger et al., 2007, 2009). On this basis a
mathematical modeling has been developed which allows
quantifying human intracranial dynamics. Thus, the vascular
system can induce a net motion of the solid brain which in
turn induces the pulsatile cerebrospinal fluid flow which can
enhance VT signal migration.
It is important to underline that while these studies give
further support to the tide hypothesis (Agnati et al., 2005a) on VT
signal migration, they can also shed light on some symptoms
present in pathologic conditions such as the hydrocephalus,
where the intrinsic brain dynamics within the cranium is altered.
3.4.
Possible interactions between transmitter spill-over
and ephaptic effects at chemical synapses
The amplitude of excitatory postsynaptic potentials and currents increases with membrane potential hyper-polarization.
This has been attributed to an increase in the driving force when
the membrane potential deviates from the equilibrium potential

of the respective ions. It has been shown that in a subset of
neocortical and hippocampal synapses postsynaptic hyperpolarization causes increased pre-synaptic release of the
transmitter.
This result is in agreement with the hypothesis of Byzov
(Byzov and Maksimov, 1998) on the existence of electrical (or
ephaptic) linking in purely chemical synapses (Voronin et al.,
1999). A new aspect to be considered is the relationship
between the electrical charge of the signals and the cell
membrane polarization and hence between ephaptic effects at
a chemical synapse and transmitter spill-over. In fact, the
electrostatic interactions between charged membranes and
charged signals may have effects on their diffusion, allowing a
larger or lower sphere of influence for perisynaptic VT signals
depending on the charge of the transmitter. It has been
shown, for instance, that the electrical field setup by AMPA
receptor-mediated excitatory currents accelerates the clearance of negatively charged glutamate molecules from the
synaptic cleft, speeding up synaptic responses and, possibly,
increasing glutamate spill-over (Sylantyev et al., 2008).
147
4.
Putative novel types of wiring and volume
transmission
As mentioned above, TNTs and exosomes may represent
novel types of WT and VT, respectively. It has to be pointed
out, however, that it is still a matter of investigation whether
TNTs exist in vivo, in particular in the brain (see below). On the
other hand, there are several findings demonstrating the
existence of exosomes (more generally of micro-vesicles, see
below) in vivo both in physiological and in pathological
conditions. However, their specific relevance for cellcell
communication, especially in the brain, is still a matter of
very intense investigations.
4.1.
Tunnelling nanotubes
Tunnelling nanotubes are structures involved in the intercellular

communication, which have been recently discovered by means of
in vitro studies (Rustom et al., 2004). In a transient way they
connect two cells forming a private direct channel, which has no
Fig. 4 Intercellular migration of mitochondria via tunnelling nanotubes (TNTs). Cortical rat astrocyte or glioblastoma (U57MG)
cell cultures were labeled by MitoTracker (MT) Green FM to visualize mitochondria and by wheat germ agglutinin (WGA) Alexa
Fluor 594 conjugate (red; plasma membrane marker) to visualize TNTs. Live cells were observed by confocal laser microscopy
and the respective images of the two stainings for astrocyte and glioblastoma cells were acquired (upper and lower panels on
the left). A discrimination procedure on the two panels was performed by means of a computer-assisted image analyzer (Zeiss
Kontron KS400). Thus, in panels A and A the discriminated images of the WGA staining for astrocyte and glioblastoma cells,
respectively, are given. Instead in panels B and B the discriminated images of the MT staining for astrocyte and glioblastoma
cells, respectively, are given. Intercellular transfer of mitochondria via TNTs was observed (see circle in panels B and B). This
image has been kindly provided by Dr. Chiara Carone (Department of Biomedical Sciences, University of Modena and Reggio
Emilia), see also Agnati et al., 2010.
148
gaps, resembling plasmodesmata of plant cells (Pontes et al., 2008).

As far as animal cells are concerned, they have been identified in a
variety of cultured cell systems, including cells of the immune
system, kidney cells, PC12 cells and cells from human glioblastoma
(see Rustom, 2009, for a comprehensive review).
TNTs have a diameter of 50200 nm and a length up to
several cell diameters. These transcellular channels could lead
to the formation of syncytial cellular networks (Rustom et al.,
2004; Onfelt et al., 2006; Gerdes and Carvalho, 2008; Sowinski et
al., 2008). As mentioned above, they are transient structures
having variable lifetimes ranging from less than 60 min (T
cells) up to several hours (PC12 cells) (Gurke et al., 2008b).
Several in vitro studies demonstrated that these structures
make possible the exchange of molecules, proteins, and whole
organelles between cells (Rustom et al., 2004; Baluka et al.,
2006; Goncharova and Tarakanov, 2008). Thus, mitochondrial
DNA as well as RNA can migrate along TNTs from one cell to
neighboring cells (Rustom et al., 2004; Koyanagi et al., 2005;
Onfelt et al., 2006; Belting and Wittrup, 2008; Agnati et al.,
2010). Furthermore, it has been shown that membrane
proteins can be exchanged unidirectionally between cells
either along the membrane of TNTs or during phases of their
transient contacts (Rustom et al., 2004; Ambudkar et al., 2005;
Watkins and Salter, 2005; Onfelt et al., 2006; Gurke et al., 2008a;
Rechavi et al., 2009). Mitochondria can also migrate unidirectionally along TNTs from one cell to neighboring cells either
inside or along their surface (see Fig. 4 and Rustom et al., 2004;
Onfelt et al., 2006; Gurke et al., 2008a,b; Vidulescu et al., 2004;
Davis and Sowinski, 2008; Agnati et al., 2010). Endosomerelated organelles like mitochondria and melanosomes are
unidirectionally transferred via actin-mediated mechanisms
at a speed of about 25 nm s1 in PC12 cells (Gurke et al., 2008b).
In addition to transfer of organelles, plasma membrane
components such as lipid-anchored proteins can laterally
transfer along TNT-like bridges into the plasma membrane of
connected cells (Gurke et al., 2008b; Agnati et al., 2010). It may

also be surmised that membrane receptors can move from the
plasma membrane of one cell to the plasma membrane of
another cell, which through this mechanism can acquire for a
transient period of time the capability to recognize and
transduce signals otherwise not recognized.
These processes may represent a novel mechanism of cell
cell communication, which allows spontaneous intercellular
transfer of cellular components (ICT).
In this context, the role of membrane juxtapositions
(Shepherd, 1994), regions of close contiguity between cell
membranes, and of transient fusions between the plasma
membranes of neighboring cells (Niu et al., 2009), has also to
be mentioned because it may contribute to ICT allowing the
simultaneous transfer of proteins, lipids and cytoplasmic
components thanks to a direct cellcell contact.
Studies carried out in co-cultures of different cell types
containing distinct levels of proteins or markers for the
plasma membrane or cytoplasm provided some support to
this possibility, indicating that a variety of trans-membrane
proteins and lipids are transferred between multiple cell types
(Koyanagi et al., 2005). In some instances also the transfer of
cytoplasmic macromolecules up to 40 kDa between somatic
cells could be demonstrated (Onfelt et al., 2006; Gousset et al.,
2009).
Thus, these mechanisms could represent an interesting
and significant type of WT, provided this mode of communication is really used by neurons and/or by the other cell types
in the CNS. In this respect, it has to be pointed out that
significant in vivo data are almost lacking, the in vivo
experimental evidence of TNTs being limited to the MHC
class II+ cells in the mouse cornea (Chinnery et al., 2008).
Therefore, future studies are required to better characterize
the properties of ICT, as well as the cell types that in vivo can
utilize this possible communication mode belonging to WT.
Table 4 Sources of the Roamer type of volume transmission (modified from Cocucci et al., 2009).
Main features
Size
Site of generation
Type of generation
Mechanism of generation
Sorting
Intracellular storage
Mechanism of discharge
Type of discharge
Typical membrane proteins
Typical lumenal proteins
Typical lipids
mtDNA transfer b
mRNA transfer
Possible physiological role
Possible pathological role
Exosomes
Shedding vesicles
30100 nm
Late endosomes (a type of MVBs)
Constitutive
Budding
Ceramide-dependent
Yes
Exocytosis of MVBs
Constitutive and regulated
CD9, CD63, GPI-anchored proteins,
TNFR1, Tsg101, Alix
Cytokines
LBPA, ceramide
Yes
Yes
Epigenesis, intercellular transfer
of packets of signals
Alzheimer's progression
100200 nm
Plasma membrane (lipid rafts?)
Regulated
Budding
Unknown
No
Budding from the plasma membranes
Regulated and possibly constitutive
Metalloproteinases, integrins
Caspase, FGF2
Cholesterol and saturated fatty acids a
Unknown
Yes
Epigenesis, coagulation, inflammation,
intercellular transfer of signalosomes
Stroke, tumor progression
Abbreviations: CD9, CD63 members of the tetraspanin family; GPI, glucosylphosphatidylinositol tail; LSBPA, 2,20-lysobisphosphatidic acid, a
phospholipid accumulated in the membrane of exosomes; MVB, multivesicular bodies; TNFR1, tumor necrosis factor receptor 1.
a
Cholesterol and saturated fatty acids can form lipid rafts, i.e., more ordered and less fluid domains than the surrounding membrane.
b
See text (Guescini et al., 2010).
4.2.
Roamer type of VT
On the basis of new data, indications have been obtained that

cells can exchange not only a single message but even a set of
messages (Fvrier and Raposo, 2004; Simons and Raposo,
2009). It has been shown that several messages can be sent via
microvesicles (acting as protective containers), dispatched
into the ECS and diffusing until the proper targets are reached
(Fig. 3). This class of intercellular communication belongs
according to our previously stated basic criterion to the VT
mode of communication (no virtually continuous channel).
However, it may be important to underline that it often shows
the important features of safety and privacy of the message.
Such a safety and privacy of the messages can only develop if
the proper target cells can recognize the micro-vesicles
though sets of adhesion molecules or receptors. The interaction may be restricted to the plasma membrane altering the
signaling of the cell or may lead to their fusion with the
plasma membrane or activation of the process of endocytosis
resulting in a decoding of the enclosed signals and in cellcell
communication as especially demonstrated in the immune
system (Lakkaraju and Rodriguez-Boulan, 2008; Cocucci et al.,
2009; Simons and Raposo, 2009).
Different types of microvesicles have been described (see
Table 4 based on Cocucci et al., 2009), which are the result of
specific cellular phenomena (for a review see Lakkaraju and
Rodriguez-Boulan, 2008). As mentioned above, the Cocucci et
al. paper (Cocucci et al., 2009) helps in having a classification,
but the field is still far from being described according to well
accepted criteria for identification of the different classes of
microvesicles. Special attention has been paid so far only to
exosomes and shedding microvesicles.
As stated in Table 4, exosomes are microvesicles
contained within a special class of endosomes. Endosomes
are membrane-bound organelles, which can be classified on
the basis of their morphology, their distinct protein and lipid
composition, their position within the cell and the cargo that
they carry. Thus, they are usually classified as early, late or
recycling endosomes and, in particular, late endosomes are a
type of multivesicular bodies (MVB), which contain intralumenal vesicles (ILVs) 30100 nm in diameter. These ILVs are
referred to as exosomes and can be released by fusion of the
limiting membrane of the MVB with the plasma membrane.
Thus, endosomes not only transport newly synthesized
material from the Golgi complex and endocytosed material
from the plasma membrane to various intracellular destinations, but an alternative fate of MVBs is their exocytic fusion
with the plasma membrane (Lakkaraju and RodriguezBoulan, 2008), leading to the release of the 40100 nm ILVs
(exosomes) into the extracellular milieu (Van Niel et al., 2006).
Exosomes are released both constitutively and in a regulated
manner (Lakkaraju and Rodriguez-Boulan, 2008). However,
the molecular machinery involved in the exocytotic fusion of
MVBs to the plasma membrane to release exosomes is still
under investigation. One hypothesis is that the regulated
release of exosomes uses similar mechanisms to those
involved in the fusion of secretory lysosomes with the
plasma membrane. Thus, exosomes are classically defined
as vesicles with a diameter of 40100 nm that originate in
MVBs of the endosomal system with an emerging role as
149
intercellular signaling devices (Barral and von Herrath, 2005).

At their limiting membrane exosomes express some markers
such as Tsg101 and CD9, which also allow their individualization-isolation (for further details see legend to Fig. 5 and
Guescini et al., 2010).
Microvesicles can also be formed from lipid raft domains of
the plasma membrane and are then called shedding vesicles
(Smalheiser, 2007). The shedding of vesicles is preceded by the
budding of small cytoplasmic protrusions, which then detach
by fission of their stalk. They are rich in cholesterol,
sphingomyelin and ganglioside GM3. Furthermore, they
express certain marker proteins (e.g., Alix and Tsg101, which
are involved in endosomal-lysosomal sorting) but lack markers of lysosomes or caveolae. Furthermore, the shedding
vesicles contain cholesterol-rich membrane rafts, with which
some proteins are selectively associated. It should be also
noted that shedding vesicles produced by different cells
contain certain proteins in common, such as those involved
in membrane trafficking (e.g., Rabs and annexins) and in MVB
biogenesis (e.g., Tsg101 and Alix), and lack proteins of the MVB
limiting membrane (Lakkaraju and Rodriguez-Boulan, 2008).
Recently, Fang et al. (2007) proposed that proteins which
exhibit higher-order oligomerization (i.e., oligomerization of
oligomers) and which also associate with the plasma membrane are preferentially sorted into the shedding vesicles.
These vesicles originating from the plasma membrane also
express specific cell-surface proteins including integrins and
cell adhesion molecules, so they have the means to bind
selectively to, and be taken up by, specific recipient cell types
having these sets of integrins and cell adhesion molecules.
The view that exosomes as well as shedding vesicles are
safe vesicular carriers for targeted intercellular communication (Simons and Raposo, 2009) is largely supported by
experimental findings. As a matter of fact, it has been
shown that intrinsic features of exosomes and of shedding
vesicles allow their specific interaction with the appropriate
target cells and to fulfill their task moving from the sourcecell to the target-cell(s) (Mallegol et al., 2007; Belting and
Wittrup, 2008; Lakkaraju and Rodriguez-Boulan, 2008;
Schorey and Bhatnagar, 2008; Simons and Raposo, 2009).
Several analogies may be proposed to illustrate this
peculiar class of VT. Lakkaraju and Rodriguez-Boulan image
exosomes to be itinerant workers who travel away from the
source of production and roam from cell to cell to disseminate
important information (Lakkaraju and Rodriguez-Boulan,
2008). Thus, we suggest calling this type of VT, the Roamer
type of VT. It is proposed that it is the vector mediated
migration in VT (see Section 3.3) that makes possible the
Roamer type of VT, namely the pressure gradients and the
temperature gradients.
Exosomes are often released as small aggregates that could
be taken up by neighboring cells via a phagocytic mechanism
(leading to signal privacy of the broadcast type). It cannot be
excluded that exosomes merely attach to the target cell
surface, conferring new properties to the cell (Fang et al.,
2007), but likely the mechanism is cell-to-cell communication
via cell surface receptors recognizing the microvesicles
(leading to signal privacy of the reserved type (see Section 1.1)).
As far as the signals conveyed to the target cell are
concerned, tissue culture studies show that exosomes can
150
Fig. 5 Biochemical characterization of glioblastoma and astrocyte exosomes (see Guescini et al., 2010). Left upper panel:
Schematic representation of the centrifugation steps used to isolate microvesicles released from glioblastoma and astrocyte
cells. Right upper panel: Mitochondrial DNA quantification from glioblastoma- and astrocyte-conditioned medium. mtDNA was
isolated from exosome pellets and quantified by real-time PCR using specific primers for the mitochondrially encoded gene
NADH dehydrogenase subunit 1 (MT-ND1). Ultracentrifugation pellet obtained from DMEM medium with addition of 10%
heat-inactivated FBS was used as a negative control. Results: DMEM + 10% FBS no detectable mtDNA quantity;
Glioblastoma = 82,700 6000; Astrocytes = 102,000 4700; the results are reported as mtDNA copy number (mean sem; n = 8).
Left lower panel: Western blot characterization of glioblastoma and astrocyte exosomes. Exosomes (Exo), supernatant (Sup)
and whole cellular lysate proteins (Lys) from glioblastoma and astrocyte cells were separated on SDS-PAGE and electroblotted to
nitrocellulose membrane. Blots were probed with antibodies against: Alix, Tsg101 and CD9. Molecular mass markers are shown
to the left. Right lower panel: Mitochondrial DNA quantification after DNaseI treatment. Intact glioblastoma-released exosomes
treated with DNaseI show a marked and significant decrease in mtDNA quantity with respect to control exosomes but about
10% of mtDNA was protected from degradation. Exogenous DNA (Exg DNA) added to the reaction was completely degraded by
DNase treatment demonstrating that complete digestion had occurred. Thus, as shown only a small fraction of mtDNA was
located inside the exosomes, and the larger fraction was present in the supernatant as naked mtDNA, which may be free in
the medium and/or associated with the external surface of the exosome membranes. Results: Exg DNA = 165 2; Exg DNA
+ DNaseI no detectable mtDNA quantity; Exosomes = 100 0.7; Exosomes + DNaseI = 9.9 0.75; Exosomes + Exg DNA
+ DNaseI = 14.5 0.75; the results are reported as percentage with respect to exosome quantity (mean sem; n = 6).
contain receptors and proteins (Lakkaraju and RodriguezBoulan, 2008; Guescini et al., 2010) (for a general panorama, see
Fig. 3). Of particular interest is some evidence indicating that
exosomes could operate as carriers for the intercellular
transfer of RNAs, given the tentative involvement of endosomal pathways in the transmission of RNAs in lower organisms
(Belting and Wittrup, 2008). The genetic material transferred to
neighboring cells via exosomes, however, does not simply
mirror the transcriptional status of the donor cell. As a matter
of fact, gene profile analysis showed significant differences in
the level of transcripts isolated from exosomes and donor

cells, which suggests that complex selection mechanisms
operate during the formation or intracellular sorting of
exosomal RNA (Ratajczak et al., 2006; Deregibus et al., 2007;
Burghoff et al., 2008). Thus, in signaling carried out by
microvesicle RNAs especially the non-coding RNAs (ncRNAs)
(Valadi et al., 2007) are of a substantial interest, in view of their
physicochemical properties, which enable these molecules to
fulfill a diverse range of structural and catalytic roles through
various mechanisms (Mehler and Mattick, 2007; Scott, 2007). It
has been suggested that several classes of ncRNAs can work

also as extracellular signaling molecules allowing to fully
exploit their ductile and fundamental capability of regulating
molecular networks (Mattick, 2007; Dinger et al., 2008; St
Laurent and Wahlestedt, 2007).
As pointed out by Dinger et al. (2008) the endosomal
machinery of signaling could be extended to include intercellular communication by taking advantage of exosomes,
which could safely deliver the RNAs to the proper target cells.
Thus, RNAs could transmit a range of signals to other cells
without having to evolve novel signal-specific pathways, a
further advantage of RNA in the context of lineage specific
developmental and physiological regulation.
Actually, according to the Jacob's tinkering principle (Jacob,
1977), it can be surmised that during evolution the endosomalmediated control, which was so successfully exploited at the
intracellular level for unicellular organisms, has been used to
allow efficient cellcell communication in multi-cellular
organisms. As a matter of fact, several data suggest that
RNAs can propagate between cells and even between organisms (Smalheiser, 2007; Dinger et al., 2008).
In the context of the present paper, however, a basic
question is the relevance of the Roamer type of VT for the
intercellular communication in the CNS.
As far as the CNS is concerned, in vitro studies showed that
neurons can release exosomes containing glutamate receptor
subunits, and that oligodendrocytes can secrete exosomes
containing myelin proteins (Lakkaraju and Rodriguez-Boulan,
2008; Guescini et al., 2010). In this context it is interesting to
notice that exosomes released from astrocytes and glioblastoma cells (Guescini et al., 2010) contain also mitochondrial DNA.
In vivo, microvesicles were identified in the human cerebrospinal fluid (Marzesco et al., 2005) and in the human and rat
CSF they appeared associated with a suite of signaling and
intracellular proteins (Zappaterra et al., 2007). These findings
open the possibility that this type of VT plays some role in the
CNS, for instance by supplementing the known mechanisms of
anterograde and retrograde signaling across synapses. In
particular, it has been suggested (Smalheiser, 2007) that
exosomes born in the plasma membrane (shedding vesicles)
bud from the lipid raft region of the postsynaptic membrane
adjacent to the postsynaptic density. As a matter of fact, Faur
et al. (2006) demonstrated that rat and mouse cortical neurons
secrete exosomes in culture. These exosomes contained two
(GluR2/3) out of the four possible AMPA receptor subunits,
which combine to form the receptor tetramer. It should be
noticed that AMPA receptors, in particular both GluR1 and
GluR2, play an important role in Long Term Potentiation
(Whitlock et al., 2006). It has been shown that homomeric
GluR1 AMPA receptors were functionally delivered to synapses
after LTP induction, whereas homomeric GluR2 or GluR3 AMPA
receptors were inserted constitutively (Whitlock et al., 2006).
Furthermore, recent evidence supports a model in which the
insertion of GluR1-containing receptors occurs indirectly via
insertion at extra-synaptic sites followed by a lateral movement into the synaptic sites (Passafaro et al., 2001). Thus, the
Roamer type of VT may greatly contribute to dynamic events of
synaptic plasticity.
Exosomal secretion could likely have other physiological
and pathological roles in CNS. In particular, two possible
151
functional roles have been suggested for microglia-derived

CNS exosomes, since they express the amino-peptidase CD13
and the lactate transporter MCT-1 (Potolicchio et al., 2005). It
can be surmised that exosomal CD13, which inactivates
leucine- and methionine-enkephalins, plays a role in the
control of these two VT signals. As a matter of fact, the two
cleaved neuropeptides are unable to bind to the neuronal
opioid receptor as assessed by the cAMP response (Potolicchio
et al., 2005). The ability of microglia-derived CNS exosomes to
carry out this process highlights our view of the CNS as an
organized system of CCNs. On the other hand, exosomal MCT1 can play a role in the metabolic support to neurons. As a
matter of fact, glycolysis and lactate transport are critical and
tightly regulated functions in brain metabolism. Neurons,
which predominantly express isoform 1 of lactate dehydrogenase, can oxidize lactate for energy production and microglia could provide lactate thanks to the MCT1 transporter
(Potolicchio et al., 2005), provided that the microvesicle
containing it can be recognized by the neuronal plasma
membrane.
As far as the possible pathological role of exosomes is
concerned, exosomes were found in a study by Vella et al.
(2008) in CSF from sheep and in these CSF-derived exosomes
the authors were able to show an enrichment of prion protein
over unfractionated CSF, suggesting a link between these
vesicles and prion disease pathogenesis. Furthermore, it has
been shown that a minute fraction of Abeta peptides can be
secreted from the cells in association with exosomes (Rajendran et al., 2006). Consistent with this finding, the presence of
exosomal proteins has been observed in plaques from
Alzheimer's disease (AD) patient brains (Rajendran et al.,
2006), supporting a potential role for exosomes in the
pathogenesis of AD (Ghidoni et al., 2008).
Taking all together, it should be considered that the
possibility of a Roamer type of VT in the CNS opens a new
research field in CNS communication of high relevance for
neurophysiology and neuropathology , allowing the exploration of new possible therapeutic interventions (Fiore et al.,
2008).
5.
On the role of the extracellular space in
intercellular communication
The intercellular space fulfils active tasks in the elaboration of
the information since it may address the diffusion of
electrochemical messages in an anisotropic fashion favoring
or preventing the communication between two brain areas
contributing to compartment formation. Furthermore, the
ECM is not an amorphous filling between the different cell
types of the CNS. On the contrary, it may affect the messages
released by the CNS cells, for example, leading to the
formation of different sets of fragments from the same parent
peptide and thus of different chemical networks that can
modulate various CCNs eliciting different types of integrated
responses (Agnati and Fuxe, 2000; Agnati et al., 2004, 2005b;
Fuxe et al., 2007b).
On this basis we proposed that the integrative tasks of the
CNS should be studied not by considering the neural networks, but whole compartments of brain tissue where
152
different cell types and ECM work as an integrated computational unit (Agnati et al., 1995c). It has also been proposed that
beside the CCNs also the molecular networks formed by
organized elements of the ECM should be considered and that
these molecular networks mainly made by proteins and
carbohydrates interact with the cell membrane molecular
networks to form a global molecular network (GMN), which
enmeshes the entire CNS (Apathy, 1897; Agnati and Fuxe,
2000; Agnati et al., 2006b; Fuxe and Agnati, 2009). Thus,
according to our hypothesis, the CNS is a computational
system with two computing nets, the CCN and the GMN,
working sometimes as parallel circuits, sometimes as serial
circuits and overlapping with each other at the cell membrane
level.
The extracellular part of GMN has both a scaffolding role in
enabling the appropriate location of the CNS components and
a functional role in cooperating to the maintenance of the
microenvironment around cells and in the creation of
preferential diffusion pathways. A model of the meshes of
the ECM has been proposed by Yamaguchi (Yamaguchi, 2000),
who suggested that hyluronan, which is one of the fundamental blocks of the ECM, can form meshes of different
tightness. In this way the hyaluronan-lectican-tenascin R
model of the brain ECM was introduced. It is obvious that
intercellular VT (i.e., signal diffusion and convection) including the Roamer type of VT is affected by the control of the net
tightness of the ECM meshes and by the medium fluidity,
which alter the tortuosity of the diffusion pathways (Nicholson et al., 2000) where VT signals including exosomes released
by the source cell migrate to reach the target cells. It should be
noticed that all the subclasses of the intercellular WT, i.e.,
synaptic contacts, gap junctions, and TNTs, also depend on
the cell circuit geometry and hence on ECM characteristics. As
a matter of fact, ECM characteristics modulate the formation
and the maintenance of these subclasses of intercellular
communication. In turn, the CCN affects the GMN molecular

composition and arrangements by secreting its components
as well as enzymes involved in its plastic rearrangements
(Rauch, 2004). Thus, a complex interplay between the molecular and topological organization of the GMN and the
intercellular WT and VT occurs to establish the optimal tuning
of the CNS integrative actions.
6.
Concluding remarks
Brain integrative action mainly depends on neural networks

and on the synaptic transmission (Sherrington, 1906), This
signaling backbone, however, is significantly complemented
by complex cellular networks (involving neurons, astrocytes,
microglial cells, oligodendroglial cells, ependymal cells).
Elements of these networks communicate basically via two
classes of connections (WT and VT). This enlarged view is in
agreement with Golgi's proposal that can be paraphrased by
stating that different brain cells (neurons and glial cells)
present in a certain brain region can operate as a single
integrative province (Golgi, 1914).
In this respect, of particular interest are, in our opinion, the
recently discovered new types of WT (i.e., TNTs) and VT (i.e.,
exosomes and shedding vesicles). Although their existence
and role in vivo in the CNS are still a matter of investigation
and remain to be fully demonstrated, the available experimental data open the possibility that the peculiar features of
these intercellular communication modes could be of paramount importance for the integrative actions of the brain
CCNs. In this context, it is interesting to cite the Smalheiser's
proposal (see Section 4.2) that exosomal transfer of proteins
and RNAs, especially from the post-synaptic dendrite to the
pre-synaptic terminal can play a role in synaptic plasticity
(Smalheiser, 2007).
Fig. 6 Exosome purification using magnetic beads (see Guescini et al., 2010). Left panel: mtDNA was quantified from
glioblastoma-released exosome fraction after purification with magnetic beads. CD9 antibodies were used in the binding of
exosomes to the beads followed by magnetic immunoprecipitation. Right panel: Quantification of mtDNA present within
purified exosomes. This figure shows that most of the pelleted mtDNA is free (SUPERNATANT) and only about 5% of the released
mtDNA is associated with exosomes (BEADS + Anti CD9). Results: SUPERNATANT = 100 6; BEADS + Anti CD9 = 6.1 1.65; the
results are reported as percentage with respect to SUPERNATANT quantity (mean sem; n = 3).
153
Fig. 7 Schematic representation of migration of mitochondria and mtDNA via WT (TNTs) and classic (regular) and Roamer type
of VT. Two possible mechanisms for mtDNA diffusion and/or flow in the extracellular fluid and hence in the liquor are
illustrated (see Fig. 8). It is surmised that mtDNA detected in the liquor of control subjects could depend on either on a release
process of mtDNA from cells and/or exosomes or on the rupture of tunnelling nanotubes with diffusion into the extracellular
space of their content including mtDNA.
It should also be mentioned that both classical VT (see

Section 3 and Figs. 5 and 6) and the Roamer type of VT (see
Section 4.2) can allow the transfer of mtDNA (as TNTs can do in
some instances, see also Fig. 4). Thus, a situation like the one
shown in Fig. 7 is very likely. Further support for such a view has
been obtained by analyzing liquor from human beings (see Fig. 8

and Fuxe et al., 2010). Hence, it is of great importance to realize
that intercellular communication can involve not only transmitters and electrical (ionic) signals, but also genetic information
(mtDNA and RNA) and even entire cell organelles (see Section 4.1),
Fig. 8 In vivo studies on mitochondrial DNA in liquor in human subjects Through mitochondrial DNA (mtDNA) quantification
from liquor of human subjects it has been shown that mtDNA may migrate in vivo as a possible VT signal. Thus, mtDNA has
been demonstrated in samples of liquors of subjects with hydrocephalus or aqueductal stenosis and in controls (Fuxe et al.,
2010). Thus, mtDNA quantification from liquor supports the view that mtDNA can operate as a VT signal. Six control subjects
(average age years = 46; sem = 5 years) affected by chronic migraine underwent to a battery of clinical investigations and also to
the examination of their liquor. Mitochondrial DNA was isolated from 200 l (microliter) of liquor using the Qiamp Mini kit
(Qiagen) according to the manufacturers instructions. Mitochondrial DNA (mtDNA) content from liquor samples was measured
by real-time PCR using specific primers for the mitochondrially encoded genes: NADH dehydrogenase subunit 1 (MT-ND1),
NADH dehydrogenase subunit 4 (MT-ND4) and cytochrome oxidase subunit 1 (MT-CO1). Mean mtDNA copy number per ml
liquor is shown in relation to controls. Aqueductal stenosis but not hydrocephalus produces a substantial and significant rise of
the mtDNA number for the three mitochondrial genes studied (p < 0.001; KruskalWallis test). These data led us to hypothesize
that higher liquor pressures may cause the rupture of tunnelling nanotubes with mitochondria leaking mtDNA into the
extracellular space.
154
leading to a transient appearance of new proteins in a cell and of

new functional properties.
As far as exosomes and shedding vesicles are concerned
(see Table 4), it should be considered that while exosomes
formed by MVB allow the transfer of packets of signals (that
is RNA, proteins, trophic factors), the shedding vesicles
formed by plasma membrane micro-domains, in particular
Lipid Rafts, may be the vehicle for transferring signalosomes
with their receptor mosaics to the target cells. In other words,
entire or parts of Horizontal Molecular Networks (Okamoto
et al., 1998; Agnati et al., 2002, 2003) can be transferred by one
cell to other cells creating (see Section 4.2) new transient
integrative complex molecular networks in cell populations.
This goal can also be the result of ICT (Niu et al., 2009; see
Section 4.1) or of TNTs since these transient transcellular
channels have the ability to traffic not only proteins but also
organelles between cells, either within their lumen or along
their surface (see above and Davis and Sowinski, 2008).
Therefore, if fully demonstrated to occur in the brain, these
novel forms of WT and VT could represent a new aspect of the
extraordinary plasticity of the CNS, offering the possibility of
new complex integrative capabilities to brain networks and
opening a new field for neuropathological investigations. As a
matter of fact, an increasing number of human diseases are
known to be caused, either directly or indirectly, by mutations in
mtDNA (Wallace et al., 1997; Wallace, 1999). In particular, the
possible roles of the mitochondrial genome in ageing (Wallace
et al., 1997) in the development of carcinogenesis (Penta et al.,
2001) and in neurodegenerative processes such as Alzheimer's
and Parkinson's disease have been demonstrated (Schapira,
2006; Ekstrand et al., 2007). Thus, altered mtDNA or mitochondria divorced from their normal endosymbiotic functions within
their host cell can become pathogenic entities involved in the
initiation and dissemination, via TNTs and/or the Roamer Type
of VT, of several pathological conditions (Agnati et al., 2010).
REFERENCES
Adams, V.L., Goodman, R.L., Salm, A.K., Coolen, L.M., Karsch, F.J.,
Lehman, M.N., 2006. Morphological plasticity in the neural
circuitry responsible for seasonal breading in the ewe.
Endocrinology 147, 48434851.
Agnati, L.F., Fuxe, K., 1996. The impact of histological techniques in
revealing brain function. Volume transmission: from fluorescence
histochemistry to confocal laser microscopy. In: Fuxe, K., Hkfelt,
T., Olson, L., Ottoson, D., Dahlstrm, A., Bjrklund, A. (Eds.),
Molecular Mechanisms of Neuronal Communication, Vol 68.
Wenner-Gren International Series, pp. 251277.
Agnati, L.F., Fuxe, K., 2000. Volume transmission as a key feature
of information handling in the central nervous system possible
new interpretative value of the Turing's B-type machine. Prog.
Brain Res. 125, 319.
Agnati, L.F., Fuxe, K., Zoli, M., Ozini, I., Toffano, G., Ferraguti, F., 1986.
A correlation analysis of the regional distribution of central
enkephalin and beta endorphin immunoreactive terminals and
of opiate receptors in adult and old male rats. Evidence for the
existence of two main types of communication in the central
nervous system: the volume transmission and the wiring
transmission. Acta Physiol. Scand. 128, 201207.
Agnati, L.F., Cortelli, P., Biagini, G., Bjelke, B., Fuxe, K., 1994.
Different classes of volume transmission signals exist in the
central nervous system and are affected by metabolic signals,

temperature gradients and pressure waves. NeuroReport 6,
912.
Agnati, L.F., Bjelke, B., Fuxe, K., 1995a. Volume versus Wiring
Transmission in the brain: a new theoretical frame for
neuropsychopharmacology. Med. Res. Rev. 15,
3345.
Agnati, L.F., Zoli, M., Strmberg, I., Fuxe, K., 1995b. Intercellular
communication in the brain: Wiring versus Volume
Transmission. Neurosci. 69, 711726.
Agnati, L.F., Cortelli, P., Pettersson, R., Fuxe, K., 1995c. The concept
of trophic units in the central nervous system. Prog. Neurobiol.
46, 561574.
Agnati, L.F., Tinner, B., Staines, W., Vnnen, K., Fuxe, K., 1995d.
On the cellular localization and distribution of carbonic
anhydrase II immunoreactivity in the rat brain. Brain Res. 676,
1024.
Agnati, L.F., Santarossa, L., Benfenati, F., Ferri, M., Morpurgo, A.,
Apolloni, B., Fuxe, K., 2002. Molecular basis of learning and
memory: modelling based on receptor mosaics. In: Apolloni, B.,
Kurfes, F. (Eds.), From Synapses to Rules. Kluwer Academic/
Plenum Publishers, New York, pp. 165196.
Agnati, L.F., Franzen, O., Ferr, S., Leo, G., Franco, R., Fuxe, K.,
2003. Possible role of intramembrane receptor-receptor
interactions in memory and learning via formation of
long-lived heteromeric complexes: focus on motor learning
in the basal ganglia. J. Neural. Transm. Suppl. 65,
128.
Agnati, L.F., Ferr, S., Leo, G., Lluis, C., Canela, E.I., Franco, R., Fuxe,
K., 2004. On the molecular basis of the receptor mosaic
hypothesis of the engram. Cell. Mol. Neurobiol. 24, 501516.
Agnati, L.F., Genedani, S., Lenzi, P.L., Leo, G., Mora, F., Ferr, S.,
Fuxe, K., 2005a. Energy gradients for the homeostatic control of
brain ECF composition and for VT signal migration:
introduction of the tide hypothesis. J. Neural Transm. 112,
4563.
Agnati, L.F., Tarakanov, A.O., Ferr, S., Fuxe, K., Guidolin, D., 2005b.
Receptor-receptor interactions, receptor mosaics, and basic
principles of molecular network organization: possible
implications for drug development. J. Mol. Neurosci. 26,
193208.
Agnati, L.F., Zunarelli, E., Genedani, S., Fuxe, K., 2006a. On the
existence of a global molecular network enmeshing the whole
central nervous system: physiological and pathological
implications. Curr. Protein Pept. Sci. 7, 315.
Agnati, L.F., Leo, G., Zanardi, A., Genedani, S., Rivera, A., Fuxe, K.,
Guidolin, D., 2006b. Volume transmission and wiring
transmission from cellular to molecular networks: history and
perspectives. Acta Physiol. (Oxf.) 187, 329344.
Agnati, L.F., Guidolin, D., Fuxe, K., 2007a. The brain as a system of
nested but partially overlapping networks. Heuristic relevance
of the model for brain physiology and pathology. J. Neural
Transm. 114, 319.
Agnati, L.F., Genedani, S., Leo, G., Rivera, A., Guidolin, D., Fuxe, K.,
2007b. One century of progress in neuroscience founded on
Golgi and Cajal's outstanding experimental and theoretical
contributions. Brain Res. Rev. 55, 167189.
Agnati, L.F., Genedani, S., Leo, G., Forni, A., Woods, A.S., Filaferro,
M., Franco, R., Fuxe, K., 2007c. Abeta peptides as one of the
crucial volume transmission signals in the trophic units and
their interactions with homocysteine. Physiological
implications and relevance for Alzheimer's disease. J. Neural.
Transm. 114, 2131.
Agnati, L.F., Guidolin, D., Baluska, F., Leo, G., Barlow, P.W., Carone,
C., Genedani, S., 2010. A new hypothesis of pathogenesis based
on the divorce between mitochondria and their host cells:
possible relevances for the Alzheimer's disease. Curr.
Alzheimer Res. (Sep 28. Electronic publication ahead
of print).
Alle, H., Geiger, J.R., 2007. GABAergic spill-over transmission onto

hippocampal mossy fiber boutons. J. Neurosci. 27, 942950.
Alvarez, J., Giuditta, A., Koenig, E., 2000. Protein synthesis in axons
and terminals: significance for maintenance, plasticity and
regulation of phenotype. With a critique of slow transport
theory. Prog. Neurobiol. 62, 162.
Ambudkar, S.V., Sauna, Z.E., Gottesman, M.M., Szakacs, G., 2005. A
novel way to spread drug resistance in tumor cells: functional
intercellular transfer of P-glycoprotein (ABCB1). Trends
Pharmacol. Sci. 26, 385387.
Apathy, S., 1897. Das leitende element des nervensystems und
sein topographischen beziehungen zu de zellen. Mittheil. Aus
der zool. Station zu Neapel. 12, 495748.
Arvanitaki, A., 1942. Effects evoked in an axon by the activity of a
contiguous one. J. Neurophysiol. 5, 89108.
Asztely, F., Erdemli, G., Kullmann, D.M., 1997. Extrasynaptic glutamate
spill-over in the hippocampus: dependence on temperature and
the role of active glutamate uptake. Neuron 18, 281293.
Bach-Y-Rita, P., 2005. Emerging concepts of brain function.
J. Integr. Neurosci. 4, 183205.
Baluka, F., Volkmann, D., Barlow, P.W., 2006. Cellcell channels
and their implications for Cell Theory. In CellCell Channels, F.
Baluka, D. Volkmann, P.W. Barlow, ed. Landes Bioscience,
Georgetown TX/Springer Science, New York, pp. 1-18.
Baraano, D.E., Ferris, C.D., Snyder, S.H., 2001. Atypical neural
messengers. Trends Neurosci. 24, 99106.
Barbour, B., 2001. An evaluation of synapse independence.
J. Neurosci. 20, 79697984.
Barral, A.M., von Herrath, M.G., 2005. Exosomes: specific
intercellular nano-shuttles? Curr. Immunol. Rev. 1, 16.
Belting, M., Wittrup, A., 2008. Nanotubes, exosomes, and nucleic
acid-binding peptides provide novel mechanisms of
intercellular communication in eukaryotic cells: implications
in health and disease. J. Cell Biol. 183, 11871191.
Bicker, S., Schratt, G., 2008. microRNAs: tiny regulators of synapse
function in development and disease. J. Cell. Mol. Med. 12,
14661476.
Bjelke, B., Fuxe, K., Agnati, L.F., 1988. Survival of
adenohypophyseal homologous transplants in the rat striatum
associated with prolactin-like immunoreactivity in the
surrounding neuropil of the striatum. Neurosci. Lett. 93,
139145.
Bloom, F., Segal, D., 1980. Endorphins in the cerebrospinal fluid. In:
Wood, J.H. (Ed.), Neurobiology of Cerebrospinal Fluid. Plenum
Press, New York.
Bloomfield, S.A., Vlgyi, B., 2009. The diverse functional roles and
regulation of neuronal gap junctions in the retina. Nat. Rev.
Neurosci. 10, 495506.
Boehning, D., Snyder, S.H., 2002. Circadian rhythms. Carbon
monoxide and clocks. Science 298, 23392340.
Brown, E.M., Gamba, G., Riccardi, D., Lombardi, M., Butters, R.,
Kifor, O., Sun, A., Hediger, M.A., Lytton, J., Hebert, S.C., 1993.
Cloning and characterization of an extracellular Ca2+-sensing
receptor from bovine parathyroid. Nature 366, 575580.
Bunin, M.A., Wightman, R.M., 1998. Quantitative evaluation of
5-hydroxytryptamine (serotonin) neuronal release and uptake:
an investigation of extrasynaptic transmission. J. Neurosci. 18,
48544860.
Bunin, M.A., Wightman, R.M., 1999. Paracrine neurotransmission
in the CNS: involvement of 5-HT. Trends Neurosci. 22,
377382.
Burbach, J.P., 1982. Neuropeptides and cerebrospinal fluid. Ann.
Clin. Biochem. 19 (Pt. 4), 269277.
Burghoff, S., Ding, Z., Gdecke, S., Assmann, A., Wirrwar, A.,
Buchholz, D., Sergeeva, O., Leurs, C., Hanenberg, H., Mller,
H.W., Bloch, W., Schrader, J., 2008. Horizontal gene transfer
from human endothelial cells to rat cardiomyocytes after
intracoronary transplantation. Cardiovasc. Res. 77, 534543.
Byzov, A.L., Maksimov, V.V., 1998. Electrical feedback in
155
chemical synapses. Ross. Fiziol. Zh. im. I. M. Sechenova 84,

10741084.
Cabello, N., Ganda, J., Bertarelli, D.C., Watanabe, M., Llus, C.,
Franco, R., Ferr, S., Lujn, R., Ciruela, F., 2009. Metabotropic
glutamate type 5, dopamine D2 and adenosine A2a receptors
form higher-order oligomers in living cells. J. Neurochem. 109,
14971507.
Cajal, S.R., 1906. The structure and connexions of neurons. Nobel
Lecture (December 12).
Celio, M.R., Spreafico, R., De Biasi, S., Vitellaro-Zuccarello, L., 1998.
Perineuronal nets: past and present. Trends Neurosci. 21,
510515.
Charles, K.J., Evans, M.L., Robbins, M.J., Calver, A.R., Leslie, R.A.,
Pangalos, M.N., 2001. Comparative immunohistochemical
localisation of GABA(B1a), GABA(B1b) and GABA(B2) subunits
in rat brain, spinal cord and dorsal root ganglion. Neuroscience
106, 447467.
Chen, K.C., Nicholson, C., 2000. Changes in brain cell shape create
residual extracellular space volume and explain tortuosity
behaviour during osmotic challenge. Proc. Natl. Acad. Sci. USA 97,
83068311.
Chinnery, H.R., Pearlman, E., McMenamin, P.G., 2008. Cutting edge:
membrane nanotubes in vivo: a feature of MHC class II+ cells in
the mouse cornea. J. Immunol. 180, 57795783.
Ciranna, L., 2006. Serotonin as a modulator of glutamate- and
GABA-mediated neurotransmission: implications in
physiological functions and in pathology. Curr.
Neuropharmacol. 4, 101114.
Clark, B.A., Cull-Candy, S.G., 2002. Activity-dependent recruitment
of extrasynaptic NMDA receptor activation at an AMPA
receptor-only synapse. J. Neurosci. 22, 44284436.
Cocucci, E., Racchetti, G., Meldolesi, J., 2009. Shedding
microvesicles: artefacts no more. Trends Cell Biol. 19, 4351.
Contreras, J.E., Snchez, H.A., Vliz, L.P., Bukauskas, F.F., Bennett,
M.V., Sez, J.C., 2004. Role of connexin-based gap junction
channels and hemichannels in ischemia-induced cell death in
nervous tissue. Brain Res. Rev. 47, 290303.
Cragg, S.J., Nicholson, C., Kume-Kick, J., Tao, L., Rice, M.E., 2001.
Dopamine-mediated volume transmission in midbrain is
regulated by distinct extracellular geometry and uptake.
J. Neurophysiol. 85, 17611771.
Davis, D.M., Sowinski, S., 2008. Membrane nanotubes: dynamic
long-distance connections between animal cells. Nat. Rev. Mol.
Cell. Biol. 9, 431436.
De Wied, D., Jolles, J., 1982. Neuropeptides derived from
pro-opiocortin: behavioral, physiological, and neurochemical
effects. Physiol. Rev. 62, 9761059.
Deitmer, J.W., Rose, C.R., 2010. Ion changes and signalling in
perisynaptic glia. Brain Res. Rev. 63, 113129.
Del Arco, A., Segovia, G., Fuxe, K., Mora, F., 2003. Changes in
dialysate concentrations of glutamate and GABA in the brain:
an index of volume transmission mediated actions?
J. Neurochem. 85, 2333.
Deregibus, M.C., Cantaluppi, V., Calogero, R., Lo Iacono, M., Tetta,
C., Biancone, L., Bruno, S., Bussolati, B., Camussi, G., 2007.
Endothelial progenitor cell derived microvesicles activate an
angiogenic program in endothelial cells by a horizontal
transfer of mRNA. Blood 110, 24402448.
Descarries, L., Mechawar, N., 2000. Ultrastructural evidence for
diffuse transmission by monoamine and acetylcholine neurons
of the central nervous system. Prog. Brain Res. 125, 2747.
Descarries, L., Seguela, P., Watkins, K.C., 1991. Nonjunctional
relationship of monoamine axon terminals in the cerebral
cortex of adult rat. In: Fuxe, K., Agnati, L.F. (Eds.), Volume
Transmission in the Brain, Novel Mechanisms for Neural
Transmission. Raven Press, New York, pp. 5362.
Descarries, L., Brub-Carrire, N., Riad, M., Bo, G.D., Mendez, J.A.,
Trudeau, L.E., 2008. Glutamate in dopamine neurons: synaptic
versus diffuse transmission. Brain Res. Rev. 58, 290302.
156
Diamond, J.S., 2001. Neuronal glutamate transporters limit

activation of NMDA receptors by neurotransmitter spillover on
CA1 pyramidal cells. J. Neurosci. 21, 83288338.
Dinger, M.E., Mercer, T.R., Mattick, J.S., 2008. RNAs as extracellular
signaling molecules. J. Mol. Endocrinol. 40, 151159.
Doly, S., Madeira, A., Fischer, J., Brisorgueil, M.J., Daval, G., Bernard,
R., Verg, D., Conrath, M., 2004. The 5-HT2A receptor is widely
distributed in the rat spinal cord and mainly localized at the
plasma membrane of postsynaptic neurons. J. Comp. Neurol.
472, 496511.
Ekstrand, M.I., Terzioglu, M., Galter, D., Zhu, S., Hofstetter, C.,
Lindqvist, E., Thams, S., Bergstrand, A., Hansson, F.S.,
Trifunovic, A., Hoffer, B., Cullheim, S., Mohammed, A.H., Olson,
L., Larsson, N.G., 2007. Progressive parkinsonism in mice with
respiratory-chain-deficient dopamine neurons. Proc. Natl.
Acad. Sci. USA 104, 13251330.
Eyman, M., Cefaliello, C., Ferrara, E., De Stefano, R., Lavina, Z.S.,
Crispino, M., Squillace, A., van Minnen, J., Kaplan, B.B., Giuditta,
A., 2007. Local synthesis of axonal and presynaptic RNA in
squid model systems. Eur. J. Neurosci. 25, 341350.
Fang, Y., Wu, N., Gan, X., Yan, W., Morrell, J.C., Gould, S.J., 2007.
Higher-order oligomerization targets plasma membrane
proteins and HIV gag to exosomes. PLoS Biol. 5, e158.
Faur, J., Lachenal, G., Court, M., Hirrlinger, J., Chatellard-Causse,
C., Blot, B., Grange, J., Schoehn, G., Goldberg, Y., Boyer, V.,
Kirchhoff, F., Raposo, G., Garin, J., Sadoul, R., 2006. Exosomes
are released by cultured cortical neurones. Mol. Cell. Neurosci.
31, 642648.
Ferr, S., Agnati, L.F., Ciruela, F., Lluis, C., Woods, A.S., Fuxe, K.,
Franco, R., 2007. Neurotransmitter receptor heteromers and
their integrative role in 'local modules': the striatal spine
module. Brain Res. Rev. 55, 5567.
Fvrier, B., Raposo, G., 2004. Exosomes: endosomal-derived
vesicles shipping extracellular messages. Curr. Opin. Cell Biol.
16, 415421.
Fiore, R., Siegel, G., Schratt, G., 2008. MicroRNA function in
neuronal development, plasticity and disease. Biochim.
Biophys. Acta 1779, 471478.
Flores, C.E., Li, X., Bennett, M.V., Nagy, J.I., Pereda, A.E., 2008.
Interaction between connexin35 and zonula occludens-1 and
its potential role in the regulation of electrical synapses. Proc.
Natl. Acad. Sci. USA 105, 1254512550.
Floresco, S.B., 2007. Dopaminergic regulation of limbic-striatal
interplay. J. Psychiatry Neurosci. 32, 400411.
Floresco, S.B., West, A.R., Ash, B., Moore, H., Grace, A.A., 2003.
Afferent modulation of dopamine neuron firing differentially
regulates tonic and phasic dopamine transmission. Nat.
Neurosci. 6, 968973.
Fuxe, K., Agnati, L.F., 1991a. Volume Transmission in the Brain,
Novel Mechanisms for Neural Transmission, Vol. 1. Raven
Press, New York.
Fuxe, K., Agnati, L.F., 1991b. Two principal modes of
electrochemical communication in the brain: volume vs.
wiring transmission. In: Fuxe, K., Agnati, L.F. (Eds.), Volume
Transmission in the Brain, Novel Mechanisms for Neural
Transmission, Vol. 1. Raven Press, New York, pp. 19.
Fuxe, K., Agnati, L.F., 2009. Cellcell communication through the
extracellular space. In: Squire, L.R. (Ed.), Encylopedia of
Neuroscience. Academic Pres, Oxford, pp. 655664.
Fuxe, K., Hkfelt, T., Eneroth, P., Gustafsson, J.A., Skett, P., 1977.
Prolactin-like immunoreactivity: localization in nerve
terminals of rat hypothalamus. Science 196, 899900.
Fuxe, K., Jansson, A., Diaz-Cabiale, Z., Andersson, A., Tinner, B.,
Finnman, U.B., Misane, I., Razani, H., Wang, F.H., Agnati, L.F.,
Ogren, S.O., 1998. Galanin modulates 5-hydroxytryptamine
functions. Focus on galanin and galanin
fragment/5-hydroxytryptamine1A receptor interactions in the
brain. Ann. N. Y. Acad. Sci. 863, 274290.
Fuxe, K., Jacobsen, K.X., Histad, M., Tinner, B., Jansson, A.,
Staines, W.A., Agnati, L.F., 2003. The dopamine D1

receptor-rich main and paracapsular intercalated nerve cell
groups of the rat amygdala: relationship to the dopamine
innervation. Neuroscience 119, 733746.
Fuxe, K., Rivera, A., Jacobsen, K.X., Histad, M., Leo, G., Horvath,
T.L., Staines, W., De la Calle, A., Agnati, L.F., 2005. Dynamics of
volume transmission in the brain. Focus on catecholamine and
opioid peptide communication and the role of uncoupling
protein 2. J. Neural Transm. 112, 6576.
Fuxe, K., Dahlstrm, A., Histad, M., Marcellino, D., Jansson, A.,
Rivera, A., Diaz-Cabiale, Z., Jacobsen, K., Tinner-Staines, B.,
Hagman, B., Leo, G., Staines, W., Guidolin, D., Kehr, J.,
Genedani, S., Belluardo, N., Agnati, L.F., 2007a. From the
Golgi-Cajal mapping to the transmitter-based characterization
of the neuronal networks leading to two modes of brain
communication: Wiring and volume transmission. Brain Res.
Rev. 55, 1754.
Fuxe, K., Canals, M., Torvinen, M., Marcellino, D., Terasmaa, A.,
Genedani, S., Leo, G., Guidolin, D., Diaz-Cabiale, Z., Rivera, A.,
Lundstrom, L., Langel, U., Narvaez, J., Tanganelli, S., Lluis, C.,
Ferr, S., Woods, A., Franco, R., Agnati, L.F., 2007b.
Intramembrane receptor-receptor interactions: a novel
principle in molecular medicine. J. Neural Transm. 114,
4975.
Fuxe, K., Ferr, S., Genedani, S., Franco, R., Agnati, L.F., 2007c.
Adenosine receptor- dopamine receptor interactions in the
basal ganglia and their relevance for brain function. Physiol.
Behav. 92, 210217.
Fuxe, K., Marcellino, D., Rivera, A., Diaz-Cabiale, Z., Filip, M., Gago,
B., Roberts, D.C., Langel, U., Genedani, S., Ferraro, L., de la Calle,
A., Narvaez, J., Tanganelli, S., Woods, A., Agnati, L.F., 2008.
Receptor-receptor interactions within receptor mosaics.
Impact on neuropsychopharmacology. Brain Res. Rev. 58,
415452.
Fuxe, K., Dahlstrm, A., Jonsson, G., Marcellino, D., Guescini, M.,
Dam, M., Manger, P., Agnati, L., 2010. The discovery of central
monoamine neurons gave volume transmission to the wired
brain. Prog. Neurobiol. 90, 82100.
Geffen, L.B., Jessell, T.M., Cuello, A.C., Iversen, L.L., 1976. Release of
dopamine from dendrites in rat substantia nigra. Nature 260,
258260.
Gerdes, H.H., Carvalho, R.N., 2008. Intercellular transfer mediated
by tunnelling nanotubes. Curr. Opin. Cell Biol. 20, 470475.
Getting, P.A., Denkin, M.S., 1985. Tritonia swimming: a model
system for integration within rhythmic motor systems. In:
Selverston, A.I. (Ed.), Model neural networks and behaviour.
Plenum Press, New York.
Ghidoni, R., Benussi, L., Binetti, G., 2008. Exosomes: the Trojan
horses of neurodegeneration. Med. Hypotheses 70, 12261227.
Goldberg, G.S., Moreno, A.P., Lampe, P.D., 2002. Gap junctions
between cells expressing connexin 43 or 32 show inverse
permeability selectivity to adenosine and ATP. J. Biol. Chem.
277, 3672536730.
Golgi, C., 1891. La rete nervosa diffusa degli organi centrali del
sistema nervoso. Suo significato fisiologico. Regio Istituto
Lombardo di Scienze e Lettere, XXIV: reprinted in french in
Archives Italiennes de Biologie, Vol. 15, pp. 434463.
Golgi, C., 1914. La moderna evoluzione delle dottrine e delle
conoscenze sulla vita, XLVII (1). Rendiconti Regio Istituto
Lombardo, Milano.
Goncharova, L.B., Tarakanov, A.O., 2008. Nanotubes at neural and
immune synapses. Curr. Med. Chem. 15, 210218.
Gonon, F., Burie, J.B., Jaber, M., Benoit-Marand, M., Dumartin, B.,
Bloch, B., 2000. Geometry and kinetics of dopaminergic
transmission in the rat striatum and in mice lacking the
dopamine transporter. Prog. Brain Res. 125, 291302.
Gonzlez, D., Gmez-Hernndez, J.M., Barrio, L.C., 2007. Molecular
basis of voltage dependence of connexin channels: an
integrative appraisal. Prog. Biophys. Mol. Biol. 94, 66106.
Goto, Y., Otani, S., Grace, A.A., 2007. The yin and yang of dopamine
release: a new perspective. Neuropharmacology 53, 583587.
Gousset, K., Schiff, E., Langevin, C., Marijanovic, Z., Caputo, A.,
Browman, D.T., Chenouard, N., de Chaumont, F., Martino, A.,
Enninga, J., Olivo-Marin, J.C., Mnnel, D., Zurzolo, C., 2009.
Prions hijack tunnelling nanotubes for intercellular spread.
Nat. Cell Biol. 11, 328336.
Greitz, D., 1993. Cerebrovascular fluid circulation and associated
intracranial dynamics, a radiographic investigation using MR
imaging and radionucleide cisternography. Thesis, University
of Stockholm.
Greitz, D., 2006. Unraveling the riddle of syringomyelia. Neurosurg.
Rev. 29, 251263.
Guescini, M., Genedani, S., Stocchi, V., Agnati, L.F., 2010. Astrocytes
and glioblastoma cells release exosomes carrying mtDNA.
J. Neural Transm. 117, 14.
Guidolin, D., Fuxe, K., Neri, G., Nussdorfer, G.G., Agnati, L.F., 2007.
On the role of receptorreceptor interactions and volume
transmission in learning and memory. Brain Res. Rev. 55,
119133.
Guillemin, R., 1978. Peptides in the brain: the new endocrinology of
the neuron. Science 202, 390402.
Gurke, S., Barroso, J.F., Gerdes, H.H., 2008a. The art of cellular
communication: tunnelling nanotubes bridge the divide.
Histochem. Cell Biol. 129, 539550.
Gurke, S., Barroso, J.F., Hodneland, E., Bukoreshtliev, N.V.,
Schlicker, O., Gerdes, H.H., 2008b. Tunnelling nanotube
(TNT)-like structures facilitate a constitutive,
actomyosin-dependent exchange of endocytic organelles
between normal rat kidney cells. Exp. Cell Res. 314, 36693683.
Haydon, P.G., 2001. Glia: listening and talking to the synapse. Nat.
Rev. Neurosci. 2, 185193.
Hensch, T.K., 2005. Critical period plasticity in local cortical
circuits. Nat. Rev. Neurosci. 6, 877888.
Herv, J.C., Bourmeyster, N., Sarrouilhe, D., Duffy, H.S., 2007. Gap
junctional complexes: from partners to functions. Prog.
Biophys. Mol. Biol. 94, 2965.
Hirrlinger, J., Hulsmann, S., Kirchhoff, F., 2004. Astroglial processes
show spontaneous motility at active synaptic terminals in situ.
Eur. J. Neurosci. 20, 22352239.
Holt, G.R., Koch, C., 1999. Electrical interactions via the
extracellular potential near cell bodies. J. Comput. Neurosci. 6,
169184.
Honor, E., 2007. The neuronal background K2P channels: focus on
TREK1. Nat. Rev. Neurosci. 8, 251261.
Hopcroft, J.E., Ullman, J.D., 1979. Introduction to Automata Theory,
Languages and Computation. Addison-Wesley.
Hrabetov, S., Hrabe, J., Nicholson, C., 2003. Dead-space
microdomains hinder extracellular diffusion in rat neocortex
during ischemia. J. Neurosci. 23, 83518359.
Jacob, F., 1977. Evolution and tinkering. Science 196,
11611166.
Jansson, A., Lippoldt, A., Mazel, T., Bartfai, T., Ogren, S.O., Sykov,
E., Agnati, L.F., Fuxe, K., 2000. Long distance signalling in
volume transmission. Focus on clearance mechanisms. Prog.
Brain Res. 125, 399413.
Jansson, A., Tinner, B., Steinbusch, H.W., Agnati, L.F., Fuxe, K.,
1998. On the relationship of 5-hydroxytryptamine neurons to
5-hydroxytryptamine 2A receptor-immunoreactive neuronal
processes in the brain stem of rats. A double immunolabelling
analysis. Neuroreport 9, 25052511.
Jansson, A., Tinner, B., Bancila, M., Verge, D., Steinbusch, H.W.,
Agnati, L.F., Fuxe, F., 2001. Relationships of 5hydroxytryptamineimmunoreactive terminal-like varicosities to
5-hydroxytryptamine-2. A receptor-immunoreactive neuronal
processes in the rat forebrain. J. Chem. Neuroanat. 22 (3), 185203.
Jansson, A., Lippoldt, A., Mazel, T., Bartfai, T., Ogren, S.O.,
Sykov, E., Agnati, L.F., Fuxe, K., 2002. Transmitter-receptor
mismatches in central dopamine, serotonin and
157
neuropeptide systems. Further evidence for volume

transmission. In: Walz, W. (Ed.), The Neuronal Environment:
Brain Homeostasis in Health and Disease. Human Press Inc,
Totowa, NJ, pp. 83108.
Kamermans, M., Fahrenfort, I., 2004. Ephaptic interactions within
a chemical synapse: hemichannel-mediated ephaptic
inhibition in the retina. Curr. Opin. Neurobiol. 14, 531541.
Kerchner, G.A., Nicoll, R.A., 2008. Silent synapses and the
emergence of a postsynaptic mechanism for LTP. Nat. Rev.
Kiss, J.P., Zsilla, G., Vizi, E.S., 2004. Inhibitory effect of nitric oxide
on dopamine transporters: interneuronal communication
without receptors. Neurochem. Int. 45, 485489.
Koyanagi, M., Brandes, R.P., Haendeler, J., Zeiher, A.M.,
Dimmeler, S., 2005. Cell-to-cell connection of endothelial
progenitor cells with cardiac myocytes by nanotubes: a
novel mechanism for cell fate changes? Circ. Res. 96,
10391041.
Krnjevic, K., 1986. Ephaptic interactions: A significant mode of
communications in the brain. News Physiol. Sci. 1, 2829.
Kuhn, T.S., 1962. The Structure of Scientific Revolutions.
University of Chicago Press, Chicago.
Kullmann, D.M., 2000. Spillover and synaptic cross talk mediated
by glutamate and GABA in the mammalian brain. Prog. Brain
Res. 125, 339351.
Kullmann, D.M., Erdemli, G., Asztely, F., 1996. LTP of AMPA and
NMDA receptor-mediated signals: evidence for presynaptic
expression and extrasynaptic glutamate spill-over. Neuron 17,
461474.
Kullmann, D.M., Ruiz, A., Rusakov, D.M., Scott, R., Semyanov, A.,
Walker, M.C., 2005. Presynaptic, extrasynaptic and axonal
GABAA receptors in the CNS: where and why? Prog. Biophys.
Mol. Biol. 87, 3346.
Lakkaraju, A., Rodriguez-Boulan, E., 2008. Itinerant exosomes:
emerging roles in cell and tissue polarity. Trends Cell Biol. 18,
199209.
Le Boudec, J.Y., Thiran, P., 2001. Network Calculus: A Theory of
Deterministic Queuing Systems for the Internet. Springer,
LNCS.
LeBeau, F.E., Traub, R.D., Monyer, H., Whittington, M.A., Buhl, E.H.,
2003. The role of electrical signaling via gap junctions in the
generation of fast network oscillations. Brain Res. Bull. 62,
313.
Lendvai, B., Vizi, E.S., 2008. Nonsynaptic chemical transmission
through nicotinic acetylcholine receptors. Physiol. Rev. 88,
333349.
Leo, G., Genedani, S., Filaferro, M., Carone, C., Andreoli, N.,
Astancolle, S., Davalli, P., Fuxe, K., Agnati, L.F., 2007.
Hyper-homocysteinemia alters amyloid peptide-clusterin
interactions and neuroglial network morphology and function
in the caudate after intrastriatal injection of amyloid peptides.
Curr. Alzheimer Res. 4, 305313.
Linninger, A.A., Xenos, M., Sweetman, B., Ponkshe, S., Guo, X.,
Penn, R., 2007. Cerebrospinal fluid flow in the normal and
hydrocephalic human brain. IEEE Trans. Biomed. Eng. 54,
291302.
Linninger, A.A., Xenos, M., Sweetman, B., Ponkshe, S., Guo, X.,
Penn, R., 2009. A mathematical model of blood, cerebrospinal
fluid and brain dynamics. J. Math. Biol. 59, 729759.
Liu, X.Y., Chu, X.P., Mao, L.M., Wang, M., Lan, H.X., Li, M.H., Zhang,
G.C., Parelkar, N.K., Fibuch, E.E., Haines, M., Neve, K.A., Liu, F.,
Xiong, Z.G., Wang, J.Q., 2006. Modulation of D2R-NR2B
interactions in response to cocaine. Neuron 52, 897909.
MacMillan, S.J.A., Mark, M.A., Duggan, A.W., 1998. The release of
b-endorphin and the neuropeptide-receptor mismatch in the
brain. Brain Res. 794, 127136.
Mallegol, J., Van Niel, G., Lebreton, C., Lepelletier, Y., Candalh, C.,
Dugave, C., Heath, J.K., Raposo, G., Cerf-Bensussan, N.,
Heyman, M., 2007. T84-intestinal epithelial exosomes bear
158
MHC class II/peptide complexes potentiating antigen

presentation by dendritic cells. Gastroenterology 132,
18661876.
Marzesco, A.M., Janich, P., Wilsh-Bruninger, M., Dubreuil, V.,
Langenfels, K., Corbeil, D., Huttner, W.B., 2005. Release of
extracellular membrane particles carrying the stem cell marker
prominin-1 (CD133) from neural progenitors and other
epithelial cells. J. Cell Sci. 118, 28492858.
Mattick, J.S., 2007. A new paradigm for developmental biology.
J. Exp. Biol. 210, 15261547.
Mehler, M.F., Mattick, J.S., 2007. Noncoding RNAs and RNA editing
in brain development, functional diversification, and
neurological disease. Physiol. Rev. 87, 799823.
Mundorf, M.L., Joseph, J.D., Austin, C.M., Caron, M.G., Wightman,
R.M., 2001. Catecholamine release and uptake in the mouse
prefrontal cortex. J. Neurochem. 79, 130142.
Nagya, J.I., Dudekb, E.F., Rashb, J.E., 2004. Update on connexins and
gap junctions in neurons and glia in the mammalian nervous
system. Brain Res. Rev. 47, 191215.
Nicholson, C., 1979. Brain cell microenvironment as a
communication channel. In: Schmitt, F.O., Worden, F.G. (Eds.),
The Neurosciences: Fourth Study Program. MIT Press,
Cambridge MA, pp. 457476.
Nicholson, C., 2000. Volume transmission in the year 2000. Prog.
Brain Res. 125, 437446.
Nicholson, C., Sykova, E., 1998. Extracellular space structure
revealed by diffusion analysis. Trends Neurosci. 21, 207215.
Nicholson, C., Chen, K.C., Hrabtov, S., Tao, L., 2000. Diffusion of
molecules in brain extracellular space: theory and experiment.
Progr Brain Res. 125, 129154.
Nieuwenhuys, R., 2000. Comparative aspects of volume
transmission, with sidelight on other forms of intercellular
communication. Prog. Brain Res. 125, 49126.
Niu, X., Gupta, K., Yang, J.T., Shamblott, M.J., Levchenko, A., 2009.
Physical transfer of membrane and cytoplasmic components
as a general mechanism of cellcell communication. J. Cell Sci.
122, 600610.
Okamoto, T., Schlegel, A., Scherer, P.E., Lisanti, M.P., 1998.
Caveolins, a family of scaffolding proteins for organizing
"preassembled signaling complexes" at the plasma membrane.
J. Biol. Chem. 273, 54195422.
Olh, S., Fle, M., Komlsi, G., Varga, C., Bldi, R., Barz, P., Tams, G.,
2009. Regulation of cortical microcircuits by unitary
GABA-mediated volume transmission. Nature 461, 12781281.
Olh, S., Komlsi, G., Szabadics, J., Varga, C., Tth, E., Barz, P.,
Tams, G., 2007. Output of neurogliaform cells to various
neuron types in the human and rat cerebral cortex. Front.
Neural Circuits 1, 17.
Oliet, S.H.R., Piet, R., Poulain, D.A., 2001. Control of glutamate
clearance and synaptic efficacy by glial coverage of neurons.
Science 292, 923926.
Oliet, S.H.R., Panatier, A., Piet, R., 2006. Functional neuronal-glial
anatomical remodelling in the hypothalamus. Novartis Found.
Symp. 276, 238248.
Onfelt, B., Nedvetzki, S., Benninger, R.K., Purbhoo, M.A.,
Sowinski, S., Hume, A.N., Seabra, M.C., Neil, M.A., French,
P.M., Davis, D.M., 2006. Structurally distinct membrane
nanotubes between human macrophages support
long-distance vesicular traffic or surfing of bacteria
J. Immunol. 177, 84768483.
Orthmann-Murphy, J.L., Abrams, C.K., Scherer, S.S., 2008. Gap
junctions couple astrocytes and oligodendrocytes. J. Mol.
Neurosci. 35, 101116.
Ostrow, L.W., Sachs, F., 2005. Mechanosensation and endothelin in
astrocyteshypothetical roles in CNS pathophysiology. Brain
Res. Brain Res. Rev. 48, 488508.
Oviedo-Orta, E., Evans, W.H., 2004. Gap junctions and
connexin-mediated communication in the immune system.
Biochim. Biophys. Acta 1662, 102112.
Paoletti, P., Ascher, P., 1994. Mechanosensitivity of NMDA

receptors in cultured mouse central neurons. Neuron 13,
645655.
Passafaro, M., Piech, V., Sheng, M., 2001. Subunit-specific temporal
and spatial patterns of AMPA receptor exocytosis in
hippocampal neurons. Nat. Neurosci. 4, 917926.
Penta, J.S., Johnson, F.M., Wachsman, J.T., Copeland, W.C., 2001.
Mitochondrial DNA in human malignancy. Mutat. Res. 488,
119133.
Petsko, G.A., Ringe, D., 2004. Protein Structure and Function. New
Science Press Ltd, London.
Piet, R., Vargova, L., Sykova, E., Poulain, D.A., Oliet, S.H.R., 2004.
Physiological contribution of the astrocytic environment of
neurons to intersynaptic crosstalk. Proc. Natl. Acad. Sci. U.S.A.
101, 21512155.
Pontes, B., Viana, N.B., Campanati, L., Farina, M., Neto, V.M.,
Nussenzveig, H.M., 2008. Structure and elastic properties of
tunnelling nanotubes. Eur. Biophys. J. 37, 121129.
Potolicchio, I., Carven, G.J., Xu, X., Stipp, C., Riese, R.J., Stern, L.J.,
Santambrogio, L., 2005. Proteomic analysis of
microglia-derived exosomes: metabolic role of the
aminopeptidase CD13 in neuropeptide catabolism. J. Immunol.
175, 22372243.
Puro, D.G., 1991. Stretch-activated channels in human retinal
Muller cells. Glia 4, 456460.
Rajendran, L., Honsho, M., Zahn, T.R., Keller, P., Geiger, K.D.,
Verkade, P., Simons, K., 2006. Alzheimer's disease
beta-amyloid peptides are released in association with
exosomes. Proc. Natl. Acad. Sci. U.S.A. 103, 1117211177.
Ramanathan, R., Basu, P., Krishnan, R., 2007. Towards a formalism
for routing in challenged networks. Proceedings of the 2nd
ACM Workshop on challenged networks. ACM, Montreal,
pp. 310.
Rash, J.E., Dillman, R.K., Bilhartz, B.L., Duffy, H.S., Whalen, L.R.,
Yasumura, T., 1996. Mixed synapses discovered and mapped
throughout mammalian spinal cord. Proc. Natl. Acad. Sci. USA
93, 42354239.
Ratajczak, J., Miekus, K., Kucia, M., Zhang, J., Reca, R., Dvorak, P.,
Ratajczak, M.Z., 2006. Embryonic stem cell-derived
microvesicles reprogram hematopoietic progenitors: evidence
for horizontal transfer of mRNA and protein delivery.
Leukemia 20, 847856.
Rauch, U., 2004. Extracellular matrix components associated
with remodeling processes in brain. Cell. Mol. Life Sci. 61,
20312045.
Rechavi, O., Goldstein, I., Kloog, Y., 2009. Intercellular exchange of
proteins: the immune cell habit of sharing. FEBS Lett. 583,
17921799.
Reichenbach, A., Derouiche, A., Kirchoff, F., 2010. Morphology and
dynamics of perisynaptic glia. Brain Res. Rev. 63, 1125.
Rice, M.E., Cragg, S.J., 2008. Dopamine spillover after quantal
release: rethinking dopamine transmission in the nigrostriatal
pathway. Brain Res. Rev. 58, 303313.
Ridet, I., Privat, A., 2000. Volume transmission. Trends Neurosci.
23, 5859.
Rivera, A., Agnati, L.F., Horvath, T.L., Valderrama, J.J., de La Calle,
A., Fuxe, K., 2006. Uncoupling protein 2/3 immunoreactivity
and the ascending dopaminergic and noradrenergic neuronal
systems. Relevance for volume transmission. Neurosci. 137,
14471461.
Rivera, A., Peafiel, A., Megas, M., Agnati, L.F., Lpez-Tllez, J.F.,
Gago, B., Gutirrez, A., de la Calle, A., Fuxe, K., 2008. Cellular
localization and distribution of dopamine D(4) receptors in the
rat cerebral cortex and their relationship with the cortical
dopaminergic and noradrenergic nerve terminal networks.
Neuroscience 155, 9971010.
Rusakov, D.A., Lehreb, K.P., 2002. Perisynaptic asymmetry of glia:
new insights into glutamate signalling. Trends Neurosci. 25,
492494.
Rustom, A., 2009. Hen or egg? Some thoughts on tunneling

nanotubes. Ann. N.Y. Acad. Sci. 1178, 129136.
Rustom, A., Saffrich, R., Markovic, I., Walther, P., Gerdes, H.-H.,
2004. Nanotubular highways for intercellular organelle
transport. Science 303, 10071010.
Saez, J.C., Berthoud, V.M., Branes, M.C., Martnez, A.D., Beyer,
E.C., 2003a. Plasma membrane channels formed by
connexins: their regulation and functions. Physiol. Rev. 83,
13591400.
Saez, J.C., Contreras, J.E., Bukauskas, F.F., Retamal, M.A., Bennett,
M.V.L., 2003b. Gap junction hemichannels in astrocytes of the
CNS. Acta Physiol. Scand. 179, 922.
Savtchenko, L.P., Rusakov, D.A., 2007. The optimal height of the
synaptic cleft. Proc. Natl. Acad. Sci. USA 104, 18231828.
Schapira, A.H., 2006. Mitochondrial disease. Lancet 368, 7082.
Schmitt, F.O., 1984. Molecular regulators of brain function. A new
view. Neuroscience 13, 9911001.
Schorey, J.S., Bhatnagar, S., 2008. Exosome function: from tumor
immunology to pathogen biology. Traffic 9, 871881.
Scimemi, A., Fine, A., Kullmann, D.M., Rusakov, D.A., 2004.
NR2B-containing receptors mediate cross talk among
hippocampal synapses. J. Neurosci. 4, 47674777.
Scott, W.G., 2007. Ribozymes. Curr. Opin. Struct. Biol. 17, 280286.
Shepherd, G.M., 1994. Neurobiology. Oxford University Press, New York.
Sherrington, C.S., 1906. The Integrative Action of the Nervous
System. Scribner, C. and Sons, New York.
Simons, M., Raposo, G., 2009. Exosomesvesicular carriers for
intercellular communication. Curr. Opin. Cell Biol. 21, 575581.
Smalheiser, N.R., 2007. Exosomal transfer of proteins and RNAs at
synapses in the nervous system. Biol. Direct. 30, 235.
Sowinski, S., Jolly, C., Berninghausen, O., Purbhoo, M.A., Chauveau,
A., Khler, K., Oddos, S., Eissmann, P., Brodsky, F.M., Hopkins, C.,
Onfelt, B., Sattentau, Q., Davis, D.M., 2008. Membrane nanotubes
physically connect T cells over long distances presenting a novel
route for HIV-1 transmission. Nat. Cell Biol. 10, 211219.
St Laurent III, G., Wahlestedt, C., 2007. Noncoding RNAs: couplers
of analog and digital information in nervous system function?
Trends Neurosci. 30, 612621.
Steinert, J.R., Kopp-Scheinpflug, C., Baker, C., Challiss, R.A.J.,
Mistry, R., Haustein, M.D., Griffin, S.J., Tong, H., Graham, B.P.,
Forsythe, I.D., 2008. Nitric oxide is a volume transmitter
regulating postsynaptic excitability at a glutamatergic
synapse. Neuron 60, 642656.
Sun, M.K., Alkon, D.L., 2002. Carbonic anhydrase gating of
attention: memory therapy and enhancement. Trends
Pharmacol. Sci. 23, 8389.
Sykova, E., 2001. Glial diffusion barriers during aging and
pathological states. Prog. Brain Res. 132, 339363.
Sykov, E., Vargov, L., 2008. Extrasynaptic transmission and the
diffusion parameters of the extracellular space. Neurochem.
Int. 52, 513.
Sykov, E., Mazel, T., Vargov, L., Vorsek, I., Prokopov-Kubinov,
S., 2000. Extracellular space diffusion and pathological states.
Prog. Brain Res. 125, 155178.
Sylantyev, S., Savtchenko, L.P., Niu, Y.P., Ivanov, A.I., Jensen, T.P.,
Kullmann, D.M., Xiao, M.Y., Rusakov, D.A., 2008. Electric fields
due to synaptic currents sharpen excitatory transmission.
Science 319, 18451849.
Szapiro, G., Barbour, B., 2007. Multiple climbing fibers signal to
molecular layer interneurons exclusively via glutamate
spillover. Nat. Neurosci. 10, 735742.
Tawse, K., 2008. Cerebrospinal fluid-tissue interactions in the
human brain. J.Y.I. 18.
Theis, M., Shl, G., Eiberger, J., Willecke, K., 2005. Emerging
complexities in identity and function of glial connexins.
Trends Neurosci. 28, 188195.
Theodosis, D.T., Poulain, D.A., 1993. Activity-dependent
neuronal-glial and synaptic plasticity in the adult mammalian
hypothalamus. Neurosci. 57, 501535.
159
Trasatti, S., 1999. 17991999: Alessandro Volta's Electric pile. Two

hundred years, but it doesn't seem like it. J. Electroanal. Chem. 460,
14.
Ungerstedt, U., Butcher, L.L., Butcher, S.G., Andn, N.E., Fuxe, K., 1969.
Direct chemical stimulation of dopaminergic mechanisms in the
neostriatum of the rat. Brain Res. 14, 461471.
Valadi, H., Ekstrm, K., Bossios, A., Sjstrand, M., Lee, J.J., Ltvall,
J.O., 2007. Exosome- mediated transfer of mRNAs and
microRNAs is a novel mechanism of genetic exchange between
cells. Nat. Cell Biol. 9, 654659.
Valiunas, V., Polosina, Y.Y., Miller, H., Potapova, I.A., Valiuniene,
L., Doronin, S., Mathias, R.T., Robinson, R.B., Rosen, M.R.,
Cohen, I.S., Brink, P.R., 2005. Connexin-specific cell-to-cell
transfer of short interfering RNA by gap junctions. J. Physiol.
568, 459468.
Van Niel, G., Porto-Carreiro, I., Simoes, S., Raposo, G., 2006.
Exosomes: a common pathway for a specialized function.
J. Biochem. 140, 1321.
Vella, L.J., Greenwood, D.L., Cappai, R., Scheerlinck, J.P., Hill, A.F.,
2008. Enrichment of prion protein in exosomes derived from
ovine cerebral spinal fluid. Vet. Immunol. Immunopathol. 124,
385393.
Vidulescu, C., Clejan, S., O'Connor, K.C., 2004. Vesicle traffic
through intercellular bridges in DU 145 human prostate cancer
cells. J Cell Mol. Med 8, 388396.
Volterra, A., Meldolesi, J., 2005. Astrocytes, from brain glue to
communication elements: the revolution continues. Nat. Rev.
Voronin, L.L., Volgushev, M., Sokolov, M., Kasyanov, A., Chistiakova,
M., Reymann, K.G., 1999. Evidence for an ephaptic feedback in
cortical synapses: postsynaptic hyperpolarization alters the
number of response failures and quantal content. Neuroscience
92, 399405.
Wallace, D.C., 1999. Mitochondrial diseases in man and mouse.
Science 283, 14821488.
Wallace, K.B., Eells, J.T., Madeira, V.M., Cortopassi, G., Jones, D.P.,
1997. Mitochondria- mediated cell injury. Symposium
overview. Fundam. Appl. Toxicol. 38, 2337.
Watkins, S.C., Salter, R.D., 2005. Functional connectivity between
immune cells mediated by tunnelling nanotubules. Immunity
23, 309318.
Whitlock, J.R., Heynen, A.J., Shuler, M.G., Bear, M.F., 2006. Learning
induces long-term potentiation in the hippocampus. Science
313, 10931097.
Witcher, M.R., Park, Y.D., Lee, M.R., Sharma, S., Harris, K.M., Kirov,
S.A., 2010. Three-dimensional relationships between
perisynaptic astroglia and human hippocampal synapses. Glia
58, 572587.
Yablonskiy, D.A., Ackerman, J.J., Raichle, M.E., 2000. Coupling
between changes in human brain temperature and oxidative
metabolism during prolonged visual stimulation. Proc. Natl.
Acad. Sci. USA 97, 76037608.
Yamaguchi, Y., 2000. Lecticans: organizers of the brain
extracellular matrix. Cell. Mol. Life Sci. 57, 276289.
Yano, S., Brown, E.M., Chattopadhyay, N., 2004. Calcium-sensing
receptor in the brain. Cell Calcium 35, 257264.
Yeager, M., Harris, A.L., 2007. Gap junction channel structure in the
early 21st century: facts and fantasies. Curr. Opin. Cell Biol. 19,
521528.
Zappaterra, M.D., Lisgo, S.N., Lindsay, S., Gygi, S.P., Walsh, C.A., Ballif,
B.A., 2007. A comparative proteomic analysis of human and rat
embryonic cerebrospinal fluid. J. Proteome Res. 6, 35373548.
Zha, X.M., Wemmie, J.A., Green, S.H., Welsh, M.J., 2006.
Acid-sensing ion channel 1a is a postsynaptic proton receptor
that affects the density of dendritic spines. Proc. Natl. Acad. Sci.
U. S. A. 103, 1655616561.
Zoli, M., Jansson, A., Sykova, E., Agnati, L.F., Fuxe, K., 1999. Volume
transmission in the CNS and its relevance for
neuropsychopharmacology. Trends Pharmacol. Sci. 20, 142150.

Understanding Wiring and Volume Transmission: Review

Enviado por

Dados do documento

Título original

Direitos autorais

Formatos disponíveis

Compartilhar este documento

Compartilhar ou incorporar documento

Opções de compartilhamento

Você considera este documento útil?

Este conteúdo é inapropriado?

Direitos autorais:

Formatos disponíveis

Understanding Wiring and Volume Transmission: Review

Enviado por

Direitos autorais:

Formatos disponíveis

B RA I N RE SE A R CH RE V I EW S 64 ( 20 1 0 ) 1 3 71 5 9

Understanding wiring and volume transmission

Accepted 17 March 2010

Available online 27 March 2010

the existence of unsuspected specialized structures for intercellular communication, such

Central nervous system

In 1986 we published our original proposal on the existence

It is our opinion that the basic dichotomic classification of

Wiring transmission and volume transmission

Taking advantage of some concepts and the lexicon offered by

illustrated in Table 1, where a possible approach (Ramanathan

if it can be altered during its pathway, as occurring, for

The specific feature of this mode of intercellular communication

The most important and well known is certainly the

Classical chemical synaptic transmission

It is beyond question that classic synaptic wiring, which

behavior. It represents the prototype of the WT since it is

Connexins (Cxs), a large family of homologous membrane

Table 1 Possible informatics criteria to classify communication networks of CNS.

Channel (Main criterion to distinguish

The whole available space between the network nodes is

permeability are under a control carried out by changes in the

Gap junctions can also be observed in chemical synapses,

Interneuronal wiring communication is certainly a basic

Static and/or dynamic a

Definitions of the terms:

Reserved: a specialized receptor apparatus is needed to decode the message

VT is characterized by the absence of any wire-like channel

Moreover, it is important to distinguish lipophilic from

Table 3 Chemical mediators involved in VT.

Mismatch between D4 IR , TH IR and

excitatory synapses. Thus, decreasing ASIC1a reduces the

Bunin and Wightman (1998)

Steinert et al. (2008)

MacMillan et al. (1998)

Fuxe et al. (1998)

rhythms and regulating its DNA binding activity (Boehning

susceptibility to degradation by extracellular RNAses (Alvarez

Ephaptic transmission or electrical VT

This was the first type of VT to be proposed to exist in the

tissue electrical resistance and reduce ephaptic transmission

Perisynaptic VT is linked to synaptic transmission and likely

2001; Piet et al., 2004), or in the indirect modulation of the

usually exhibits a low signal safety (Table 2 and Agnati et al.,

the membrane potential deviates from the equilibrium potential

Tunnelling nanotubes are structures involved in the intercellular

gaps, resembling plasmodesmata of plant cells (Pontes et al., 2008).

connected cells (Gurke et al., 2008b; Agnati et al., 2010). It may

On the basis of new data, indications have been obtained that

intercellular signaling devices (Barral and von Herrath, 2005).

the level of transcripts isolated from exosomes and donor

has been suggested that several classes of ncRNAs can work

functional roles have been suggested for microglia-derived

communication. In turn, the CCN affects the GMN molecular

Brain integrative action mainly depends on neural networks

It should also be mentioned that both classical VT (see

been obtained by analyzing liquor from human beings (see Fig. 8

leading to a transient appearance of new proteins in a cell and of

central nervous system and are affected by metabolic signals,

Alle, H., Geiger, J.R., 2007. GABAergic spill-over transmission onto

chemical synapses. Ross. Fiziol. Zh. im. I. M. Sechenova 84,

Diamond, J.S., 2001. Neuronal glutamate transporters limit

Staines, W.A., Agnati, L.F., 2003. The dopamine D1