URINE FORMATION AND

MEASUREMENT
 LO
Defined ginjal function
Defined urine formation
Defined how to make dilute and
concentrate urine
Defined renal clearance
KIDNEY FUNCTIONS
 URINE FORMATION

Tortora et al, 2009

Glomerular filtration
The filtration
membrane
Fenestration
(pore) of
glomerular
endothelial cell
Basal lamina of
glomerulus
Slit membrane
between
pedicels
 Net Filtration Pressure

Tortora et al, 2009

 GLOMERULAR FILTRATION
RATE
The amount of filtrate formed in all the renal
corpuscles of both kidneys each minute
+ 125 mL/min in males and 105 mL/min in
females.
Homeostasis of body fluids requires that the
kidneys maintain a relatively constant GFR.
If the GFR is too high, needed substances may
pass so quickly through the renal tubules that
some are not reabsorbed and are lost in the urine.
If the GFR is too low, nearly all the filtrate may be
reabsorbed and certain waste products may not
be adequately excreted.
The basic mechanisms that
regulate GFR
Renal Regulation of GFR
 TGF

Tortora et al, 2009

 TUBULAR REABSORPTION
Reabsorptionthe return of most of the
filtered water and many of the filtered solutes
to the bloodstream
About 99% of the filtered water is
reabsorbed.
Epithelial cells all along the renal tubule and
duct carry out reabsorption, but proximal
convoluted tubule cells make the largest
contribution.
Solutes that are reabsorbed by both active
and passive processes include glucose,
amino acids, urea, and ions (Na, K, Ca2+, Cl,
HCO3 and HPO4)
 TUBULAR SECRETION
The transfer of materials from the blood and
tubule cells into tubular fluid.
Secreted substances include ions (H, K, NH4
creatinine, and certain drugs such as
penicillin)
Tubular secretion has two important
outcomes:
(1) The secretion of H helps control blood pH.
(2) The secretion of other substances helps
eliminate them from the body.
 REABSORPTION ROUTE
Two route :
Paraceluller :
moving
between tubule
cells
Transeluller :
passing
through a
tubule cell

Tortora et al, 2009

 Transport Mechanisms
Primary active transport the energy derived
from hydrolysis of ATP is used to pump a
substance across a membrane; the sodium
potassium pump is one such pump.
Secondary active transport the energy stored
in an ions electrochemical gradient, rather than
hydrolysis of ATP, drives another substance across
a membrane.
- Symporters are membrane proteins that move
two or more substances in the same direction
across a membrane.
- Antiporters move two or more substances in
opposite directions across a membrane.
Water Reabsorption
 Reabsoption and Secretion

in PCT
Most solute reabsorption in the proximal
convoluted tubule (PCT) involves Na.
Na transport occurs via symport and
antiport mechanisms in the proximal
convoluted tubule.
Normally filtered glucose, AA, lactid acid,
water-soluble vitamin are completely
reabsorbed in the first half of the proximal
convoluted tubule by several types of Na
symporters located in the apical
membrane
Reabsorption of glucose by Na-
glucose
symporters in cells of PCT

Tortora et al, 2009

 In another secondary active transport
process, the Na/H antiporters carry
filtered Na down its concentration
gradient into a PCT cell as H is moved
from the cytosol into the lumen
causing Na to be reabsorbed into blood
and H to be secreted into tubular fluid.
PCT cells produce the H needed to
keep the antiporters running in the
following way.
Tortora et al, 2009
Tortora et al, 2009
 Solute
reabsortion in
PCT promotes
osmosis of water
by transleluler
and paraceluller
routes
PCT and
ascending loop
cell very
permeable to
water because
Tortora et al, 2009 Aquaporin-1
 REABSORPTION IN LOOP
HENLE
Tubular composition quite different with
glomerular filtrate because glucose, AA and
other nutrients are no longer presents
Reabsorbs :
15% of filtered water
20-30% of filtered Na and K
35% of filtered Cl
10-20% of filtered HCO3
and a variable amount of filtered Ca and Mg
Descending limbs reabsorb 15% of water, but
ascending limbs is impermeable to water
 REABSORPTION IN LOOP
HENLE (THICK ASCENDING)
Main effect
of Na+-K+-
2Cl-
sympoters
is
reabsoption
of Na and
Cl

Tortora et al, 2009

 REABSORPTION IN THE
Reabsorb
EARLY DCT
10-15% of filtered water
5% of filtered Na+
5% of filtered Cl-
Reabsorption Na and Cl by Na+ Cl- symporter
in the apical membran
Reabsorption Na and Cl by Sodium-potasium
pumps and Cl leakage channel in basolateral
membrans
Major site where PTH stimulates reabsorption
of Ca
REABSORPTION AND SECRETION IN
THE LATE DCT AND COLLECTING
DUCT
There are two type cell :
principal cell : reabsorb Na+, secrete
K+
intercalatus cell : reabsorb K+ and
HCO3-, secrete H+
The amount of water and solute
reabsorption and secretion vary
depending on the bodys needs
Tortora et al, 2009
RESUME
Despospolulus et al, 2003
 HORMONAL REGULATION OF
TUBULAR REABSORPTION AND
SECRETION
FORMATION OF DILUTE AND
CONCENTRATE URINE
Eventhough our fluid intake can
highly variable, the total volume of
fluid in our body normally remains
stable.
Highly fluid intake large volume of
dilute urine
Low fluid intake or loss of fluid
small volume of concentrate urine
PRODUCTION OF DILUTE
URINE

Tortora et al, 2009

 PRODUCTION OF
CONCENTRATE URINE
When water intake is low or water loss is
high (such as during heavy sweating), the
kidneys must conserve water while still
eliminating wastes and excess ions
Under the influence of ADH, the kidneys
produce a small volume of highly
concentrated urine.
Urine can be four times more concentrated
(up to 1200 mOsm/liter) than blood plasma
or glomerular filtrate (300 mOsm/liter).
 Two main factors contribute to building
and maintaining this osmotic gradient:
(1) differences in solute and water
permeability and reabsorption in
different sections of the long loops of
Henle and the collecting ducts,
(2) the countercurrent flow of fluid
through tube-shaped structures in the
renal medulla
Two types of countercurrent
mechanisms exist in the kidneys:
countercurrent multiplication and
countercurrent exchange
COUNTERCURRENT
MECHANISM
Tortora et al, 2009
The Countercurrent
Multiplier System
Extrusion of NaCl from
ascending limb makes
surrounding interstitial
fluid more concentrated
Concentration multiplied
due to descending limb
Passively permeable to H 2O
Fluid concentration
increases
As surrounding interstitial
fluid becomes more
concentration

Deepest region of
medulla
1,400mOsm
Ascending Limb Loop
of Henle
Thin segment in
depths of medulla and
thick segment toward
cortex
Impermeable to H2O;
permeable to salt
Thick segment Actively
Transports NaCl out of
filtrate

Active Transport of
salt
filtrate becomes dilute
(100 mOsm) by end of
Loop of Henle
Active Transport in
Ascending Limb
Na+ actively transported across basolateral
membrane by Na+/ K+ pump
Cl- passively follows Na+ down electrical gradient
K+ passively diffuses back into filtrate
Transport of Ions in Ascending
Limb
In thick segment Na and K together with 2 Cl- enter tubule
+ +

cells
Na+ then actively transported out into interstitial space, Cl -
follows passively
K+ diffuses back into filtrate; some also enters interstitial space
 Vasa Recta
For countercurrent multiplier system to be effective:
Most of the salt extruded from ascending limbs must
remain in the interstitial fluid of the medulla
Most of the water that leaves descending limbs must be
removed by the blood

This is accomplished by the vasa recta

Thin-walled capillaries parallel LH of juxtamedullary
nephrons
Walls permeable to water because of aquaporins
channels, NaCl, and urea but not plasma proteins
Therefore colloid osmotic (oncotic) pressure in vasta recta
is higher than in surrounding tissue fluid
results in movement of H2O from interstitial fluid into ascending
vasa recta that can be removed from the renal medulla
 Countercurrent Exchange in
Vasa Recta
Important component of
countercurrent multiplier
Permeable to salt, H2O
(via aquaporins), and urea
Recirculates salt, trapping
some in medulla
interstitial fluid (maintains
hypertonicity)
Reabsorbs H2O coming
out of descending limb
Descending section has
urea transporters
Ascending section has
fenestrated capillaries
Tortora et al, 2009
 Effects of Urea
Urea
contributes to
high osmolality
in medulla
Deep region of
collecting duct
is permeable to
urea &
transports it
The Role of Urea in Urine
Concentration
1. Urea diffuses out of
inner collecting duct
(in renal medulla)
into interstitial fluid
2. Urea then passes
into ascending limb
Recirculates in
interstitial fluid of
renal medulla
Urea and NaCl in
interstitial fluid make
it very hypertonic, so

3. Water leaves the

CD by osmosis
 Osmolality of Different Regions of
the Kidney
Countercurrent
multiplier system in LH
and countercurrent
exchange in vasa recta
Creates hypertonic renal
medulla

Under influence of ADH

CD becomes more
permeable to H2O
Thus more H2O is drawn
out by osmosis into
hypertonic renal medulla
and peritubular
capillaries
Osmotic Gradient in the Renal
Medulla

Figure 25.13
Urine Concentrating
Mechanisms
 Summary Counter-Current
Peritubular fluid
Multiplier
1 2 3 4
Tubular fluid Fluid
300 300 300 300 300 300 300 300 300 300 300 300
Cortex

Medulla 300 300 300 300 400 200 400 400 200 300 400 200

Na + H2O Na +

Cl Cl
300 400 200 400 400 200
300 300 300 300 400 200
No osmotic H2O
K+ K+
gradient
(mOsm) 300 400 200 400 400 200
300 300 300 400 400 400

300 300 300 300 400 200 400 400 200 400 400 400

Fluid Active transport Water moves Iso-osmotic state in More fluid Active
enters of Na+, Cl, K+ ions out of descending descending limb; enters tubule, transport of
tubule into medullary limb by osmosis osmotic difference pushing fluid Na+, Cl, K+
interstitial fluid between descending through by ions into
increases and ascending limbs bulk flow medullary
osmolarity interstitial
fluid increases
osmolarity

5 6 7

300 300 200 300 300 200 300 300 100

300 300 100

300 350 150 350 350 150
H2O
500 500 300
300 350 150 350 350 150 Osmotic
gradient
H2O established 900 900 700
400 500 300 500 500 300 (mOsm)
H2O 1200 1200 1100

1400
400 500 300 500 500 300
1400

Water moves Iso-osmotic state in descending More water System

out of descending limb; osmotic difference enters tubule is in steady
limb by osmosis between descending and and process state
ascending limbs continues
RENAL CLEARANCE
 Urinary Excretion of Solutes
Sometimes tricky to calculate.
Things like univalent electrolytes, glucose, aas
are freely filterable.
However, if solute binds to protein, for example,
then its not (e.g. Ca2+, PO42-, Mg2+, PAH).
For these, you must measure plasma binding
and correct for the non-filterable fraction of
solute.
Solute must also not be synthesized or
degraded.
Ammonium is synthesised, and
glutamate/glutamine is degraded, as are other
organic acids.
There are also complex combinations of
reabsorption and secretion with K +, uric acid and
urea.
Must be careful, otherwise you will get
inaccurate measurements.

Urinary
Filtered Reabsorption Secretion by
Excretion = Load - by Tubules + Tubules
of Solute
Filtration, reabsoption, and excretion rates of substances by the kidneys

Filtered Reabsorbed Excreted Reabsorbed

(meq/24h) (meq/24h) (meq/24h) (%)

Glucose (g/day) 180 180 0 100

Bicarbonate (meq/day) 4,320 4,318 2 > 99.9
Sodium (meq/day) 25,560 25,410 150 99.4
Chloride (meq/day) 19,440 19,260 180 99.1
Water (l/day) 169 167.5 1.5 99.1
Urea (g/day) 48 24 24 50
Creatinine (g/day) 1.8 0 1.8 0
 Introduction to Renal
Measurement
There are 2 main categories of tests for
assessing renal function.
1. Modern imaging techniques macroscopic
views of renal blood flow, filtration and
excretory function.
2. Measurements of renal clearance of
various substances to evaluate the ability
of the kidneys to handle solutes and water.
 Glomerular filtration is the flow of fluid
from the glomerular capillaries into the
Bowmans capsule. The volume filtrate
formed per unit time is called
glomerular filtration rate
The rate of glomerular filtration averages
135-180L/day in a normal adult.
 What is Clearance?
The clearance of a substance is the volume of
plasma from which that substance is cleared by the
kidneys per unit time. Each substance in the
plasma has its own distinct clearance value but it is
always in the form of volume of plasma per time.
To measure GFR
Substance able to achieve constant plasma
concentration
Freely filtered at the glomerulus
Not reabsorbed or secreted
Filtered substance (eg inulin)= excreted substance
GFR x Pin = Uin x V
Use creatinine clearance as estimate of GFR
Creatinine clearance=Ucr x V/Pcr
Substance X is filtered and secreted but not reabsorbed.
Substance Y is filtered and some of it is reabsorbed.
Substance Z is filtered and completely reabsorbed.
 Measurement of GFR
GFR also assessed using principles of
clearance.
GFR = vol. of fluid filtered into Bowmans
capsule per unit time.
Same equation, GFR is Cx if X has certain
required properties (i.e. Cinulin).

Conc. of X in urine
Volume of urine
formed in given

time
GFR = Ux x V
Glomerular Conc. of X in
filtration rate Px systemic blood
plasma
 Solutes used to measure GFR
Required properties are:
1. Solute is freely-filtered (conc. in Bowmans
space = that in blood plasma).
2. Tubules do not absorb, secrete or metabolize
X.
Thus, amount of X in urine per unit time
= that which glomerulus filters per unit
time.
WHAT GOES IN = WHAT COMES OUT!
INULIN is such a substance that satisfies
all of these criteria and is commonly used
to measure GFR.
 Qualities of agents to measure GFR
Inulin: (Polysaccharide from Dahalia
plant)
Freely filterable at glomerulus
Does not bind to plasma proteins
Biologically inert
Non-toxic, neither synthesized nor
metabolized in kidney
Neither absorbed nor secreted
Does not alter renal function
Can be accurately quantified
Low concentrations are enough (10-20
mg/100 ml plasma)
 Drawbacks of Inulin
Most reliable method of measuring GFR, not
useful clinically.
Inulin must be administered by IV to get relatively
constant plasma levels.
Chemical analysis of inulin in plasma and urine is
technically demanding.
Use radiolabelled compounds instead like
radioactive Vitamin B or EDTA.
However, these may also bind to proteins and
distort results slightly.
Problems of IV infusion of GFR marker avoided by
using an endogenous substance with inulin-like
properties CREATININE.
Renal handling of inulin

Amount filtered = Amount excreted

Pin x GFR Uin x V
Inulin Clearance
 Renal Plasma Flow
PAH delivery to kidney=PAH
excretion
ERPF x P PAH =U PAH x V

Note: PAH only 85-90% is removed by

kidney on single pass, thus it will
underestimate renal plasma flow
(filtered and secreted)
Measurement of Renal Blood Flow

Not all blood delivered to glomerulus is

filtered into glomerular capsule
20% is filtered; rest passes into efferent arteriole
and back into circulation
Substances that aren't filtered can still be cleared
by active transport (secretion) into tubules
 p-aminohippurate (PAH)
There are certain special cases where the
kidneys completely clear X from plasma
during a SINGLE PASSAGE through them.
In this case, renal clearance of X =
arterial renal plasma flow.
p-aminohippurate (PAH) is such a special
solute.
Thus, PAH clearance is a good estimate of
renal plasma flow.
 Renal handling of PAH
PAH is an organic acid
that is not usually
present in the body, so
must give by IV
infusion.
Note that there is none
left in the renal vein - all
cleared in first pass.
Drawback is that
mustnt give too much
PAH, otherwise we
overwhelm the PAH
secretory system and
the data can be
misleading.
 Use of PAH Clearance to Estimate Renal Plasma Flow

Paraminohippuric acid (PAH) is freely filtered and secreted

and is almost completely cleared from the renal plasma

1. amount enter kidney =

RPF x PPAH

2. amount entered =~ amount excreted

3. ERPF x Ppah = UPAH x V
~ 10 % PAH
ERPF = UPAH x V remains
PPAH
ERPF = Clearance PAH
 Creatinine clearance
Creatinine is an end product of
muscle metabolism
Muscle mass is constant; creatinine
is constant
Normal 0.7 1.5 mg/ dL in plasma
Can then be compared to creatinine
in urine over 24 hour period to
determine clearance
74
 Creatinine clearance is an indirect
measure of GFR and renal blood flow
Creatinine is neither reabsorbed nor
secreted, just freely filtered.
Amount excreted = amount filtered
Useful to monitor changes in chronic
renal function
Increases with trauma with massive
muscle breakdown

75
TERIMA KASIH