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Comparative Biochemistry and Physiology, Part A 149 (2008) 109 – 128

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Review
Ghrelin: A multifunctional hormone in non-mammalian vertebrates
Hiroyuki Kaiya a,⁎, Mikiya Miyazato a , Kenji Kangawa a , Richard E. Peter b , Suraj Unniappan c,⁎
a
Department of Biochemistry, National Cardiovascular Center Research Institute, Osaka, Japan
b
Department of Biological Sciences, University of Alberta, Edmonton, Alberta, Canada
c
Department of Biology, York University, Toronto, Ontario, Canada
Received 31 October 2007; received in revised form 3 December 2007; accepted 4 December 2007
Available online 14 December 2007

Abstract

In mammals, ghrelin is a non-amidated peptide hormone, existing in both acylated and non-acylated forms, produced mainly from the X/A or
ghrelin cells present in the mucosal layer of the stomach. Ghrelin is a natural ligand of the growth hormone (GH) secretagogue-receptor (GHS-R),
and functions primarily as a GH-releasing hormone and an orexigen, as well as having several other biological actions. Among non-mammalian
vertebrates, amino acid sequence of ghrelin has been reported in two species of cartilaginous fish, seven species of teleosts, two species of
amphibians, one species of reptile and six species of birds. The structure and functions of ghrelin are highly conserved among vertebrates. This
review presents a concise overview of ghrelin biology in non-mammalian vertebrates.
© 2007 Elsevier Inc. All rights reserved.

Keywords: Amphibians; Birds; Brain; Fish; Food intake; Ghrelin; Gut; Hormones; Peptides; Reptiles

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 110
2. From growth hormone secretagogues to ghrelin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 110
3. Ghrelin — from gene to peptide . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 111
3.1. Ghrelin gene. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 111
3.2. Ghrelin precursor, mature peptide and obestatin. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 111
3.2.1. Ghrelin precursor . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 111
3.2.2. Mature peptide . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 111
3.2.3. Acylation at the third amino acid . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 113
3.2.4. Active core of ghrelin molecule . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 113
3.2.5. Des-acyl ghrelin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 113
3.2.6. Obestatin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 113
4. Tissue distribution and localization of ghrelin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 113
4.1. Stomach . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 115
4.2. Brain. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 116
4.3. Ghrelin levels in plasma or tissues . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 116
5. Ghrelin receptor (GHS-R) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 116

⁎ Corresponding authors. H. Kaiya is to be contacted at Department of Biochemistry, National Cardiovascular Center Research Institute, 5-7-1 Fujishirodai, Suita,
Osaka 565-8565, Japan. Tel.: +81 6 6833 5012x2458; fax: +81 6 6835 5402. S. Unniappan, Department of Biology, York University, 4700 Keele Street, Toronto,
Ontario, Canada M3J 1P3.
E-mail addresses: kaiya@ri.ncvc.go.jp (H. Kaiya), suraju@yorku.ca (S. Unniappan).

1095-6433/$ - see front matter © 2007 Elsevier Inc. All rights reserved.
doi:10.1016/j.cbpa.2007.12.004
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110 H. Kaiya et al. / Comparative Biochemistry and Physiology, Part A 149 (2008) 109–128

6. Biological actions of ghrelin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 119

6.1. Regulation of hormone release and synthesis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 119
6.1.1. Fish . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 119
6.1.2. Amphibians . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 120
6.1.3. Birds . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 121
6.1.4. Reptiles . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 121
6.2. Appetite regulatory effects . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 121
6.2.1. Fish . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 121
6.2.2. Amphibians . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 122
6.2.3. Birds . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 122
6.2.4. Reptiles . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 122
6.3. Reproduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 122
6.3.1. Fish . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 122
6.3.2. Birds . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 122
6.4. Gastrointestinal motility . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 122
6.5. Regulation of water intake . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 123
6.6. Immunity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 123
6.7. Growth effects . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 123
6.8. Behavior . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 123
7. Perspectives . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 124
8. Summary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 124
Acknowledgments . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 124
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 124

1. Introduction in vitro in chicken (Bowers et al., 1984; Geris et al., 1998;

Gut hormones regulate a myriad of physiological functions
including appetite and energy balance in vertebrates (Badman
and Flier, 2005). Ghrelin, the only known appetite-stimulatory
(orexigenic) gut hormone, is an acylated peptide that was
recently added to the growing list of gut regulatory peptides
(Kojima et al., 1999). Ghrelin is involved in various physiolo-
gical functions other than GH release and appetite in mammals
(Korbonits et al., 2004; van der Lely et al., 2004; Kojima and
Kangawa, 2005; Hosoda et al., 2006). There are several lines of
evidence for the existence, wide distribution and multiple
biological actions of ghrelin in non-mammalian vertebrates, as
well as in mammals. This review focuses on a comparative
discussion of ghrelin in fish, amphibians, reptiles and birds with
mammals in terms of its structure, distribution and functions.
2. From growth hormone secretagogues to ghrelin
Growth hormone secretagogues (GHS) are synthetic com-
pounds having growth hormone (GH)-releasing actions (Tan-
nenbaum and Bowers, 2001). Some of these GHS also have
appetite-stimulatory effects in mammals (Laferrere et al., 2005).
GHS have been used in several studies on the regulation of GH
release or feeding in non-mammalian vertebrates. Intraperito-
neal (IP) injections of a GHS, KP-102, increased circulating GH
levels in Nile tilapia, Oreochromis niloticus (Shepherd et al.,
2000). However, other GHS, GHRP-6 and hexarelin, were
found ineffective in stimulating GH release in vitro and in vivo
in a grass carp, Ctenopharyngodon idellus (Xiao et al., 2002),
suggesting a species-specific action of GHS. Synthetic GHS, L-
692,429 and L-163,255 stimulate the release of GH in vivo and
2001). Geris et al. (2001) also reported that GHS stimulates the
release of an endogenous GH-releasing hormone, thyrotropin-
releasing hormone (TRH) from chicken brain. These results
indicate a direct effect of GHS on the hypothalamus as well as
its role in the pituitary to regulate GH release in chicken. The
results observed in fish and birds are in agreement with the
hypophysiotropic functions of GHS in mammals (Tannenbaum
and Bowers, 2001). Furthermore, GHS participates in the
regulation of food intake. Saito et al. (2002a) reported that
intracerebroventricular (ICV) injections of GHRP-6 reduce
food intake in neonatal chicks. Effects of GHS, although studied
only in a limited number of non-mammals, suggest the potential
presence of an endogenous GHS and its receptor in non-
mammalian vertebrates.
In 1996, GHS were found to bind to an orphan receptor
discovered from human and swine. Binding of GHS elicited an
increase in intracellular calcium in cells stably expressing the
orphan receptor. This finding led to the naming of the receptor
as the growth hormone secretagogue-receptor 1a (GHS-R1a)
(Howard et al., 1996). An inactive alternative splice variant of
the GHS-R subtype, termed GHS-R1b, has also been found,
although the function is unknown (Howard et al., 1996). Many
groups explored the endogenous ligand for the GHS-R1a,
Kojima and colleagues succeeded to de-orphanize the GHS-R1a
by discovering its endogenous ligand from rat and human
stomach (Kojima et al., 1999). The identified peptide is 28-
amino acid long and has a unique fatty acid modification at the
N-terminal third amino acid, serine (Ser-3) (Fig. 1). The peptide
has been named “ghrelin”, a term derived from the Proto-Indo-
European word “ghre” meaning “grow” and the name can also
indicate the abbreviation for GH, followed by “relin” a suffix
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H. Kaiya et al. / Comparative Biochemistry and Physiology, Part A 149 (2008) 109–128
Fig. 1. Primary structure of ghrelin in human and eel. Human ghrelin is a non-
amidated 28-amino acid peptide. Eel ghrelin is a 21-amino acid peptide and has
an amide structure at the C-terminal end.
meaning releasing substance. The fascinating story of ghrelin
discovery has been described in some reviews (Kojima et al.,
2001a; 2001b; Kojima and Kangawa, 2005). The International
Union of Pharmacology Committee on Receptor Nomenclature
and Drug Classification has accepted “GHS-R1a” as the name
for the ghrelin receptor and “GRLN” as the abbreviation for
ghrelin (Davenport et al., 2005).
3. Ghrelin — from gene to peptide
Since ghrelin was discovered in rats and humans in 1999, the
existence of ghrelin has been demonstrated in different non-
mammalian vertebrate species from cartilaginous fish to birds
(Table 1).
3.1. Ghrelin gene
It has been reported that human ghrelin gene consists of four
exons and three introns (Wajnrajch et al., 2000). On the other
hand, in rats and mice, an additional 19-bp non-coding short
exon was present (Tanaka et al., 2001) and rodent ghrelin gene
results in five exons and four introns. Recently, the presence of
an additional 20-bp non-coding exon in human ghrelin gene
was discovered after detailed sequence analyses. Based on this
new data, the human ghrelin gene is composed of five exons and
four introns (Wei et al., 2005).
In non-mammalian vertebrates, sequence of the ghrelin gene
and exon:intron organization have been reported in goldfish
(Unniappan et al., 2002), Nile tilapia (Parhar et al., 2003),
rainbow trout (Kaiya et al., 2003c), seabream (Yeung et al.,
2006), chicken (Nie et al., 2004; Richards et al., 2006) and
turkey (Richards et al., 2006) (Table 1). The ghrelin gene in
goldfish, tilapia and seabream has four exons and three introns.
The sequence identity of the short, non-coding 1st exon found
in humans, rats and mice ghrelin gene is currently not evident in
goldfish, tilapia and seabream. On the other hand, the ghrelin
gene of rainbow trout (Kaiya et al., 2003c), chicken (Nie et al.,
2004; Richards et al., 2006) and turkey (Richards et al., 2006) is
111
comprised of five exons and four introns as seen in rats, mice
and humans. The ghrelin gene organization of a reptile, the red-
eared slider turtle has been identified and it is composed of five
exons and four introns (H. Kaiya, unpublished data). The
ghrelin gene organization in amphibians is currently unknown.
3.2. Ghrelin precursor, mature peptide and obestatin
3.2.1. Ghrelin precursor
Fig. 2 shows a comparison of prepro-ghrelin sequences from
various non-mammalian vertebrates. Ghrelin precursor is
composed of a signal peptide, ghrelin mature peptide and the
C-terminal peptide (C-peptide). Length of ghrelin precursor
protein varies from species to species and it is 103-amino acid
long in goldfish, while only contains 124 amino acids in turtle.
Mature peptide is processed from the precursor protein. A
dibasic processing sequence (arginine (Arg)-Arg) is present at
the C-terminal end of mature ghrelin with some exceptions:
turkey has a proline (Pro)-Arg sequence, and the Perciformes,
including tilapia, sea bass, seabream and grouper have a single
Arg (Fig. 2). In addition to these, all teleost ghrelin has an amide
structure at the C-terminus of the mature peptide (Fig. 1). In
contrast, tetrapod ghrelin including amphibians, reptiles and
mammals do not have the structure (Fig. 1). A glycine residue
existing at the C-terminus of the mature peptide after processing
contributes to the formation of the amide structure. Presence of
non-amidated, glycine (Gly)-extended ghrelin has also been
reported in rainbow trout (Kaiya et al., 2003c). Although the
mature peptide is processed at the dibasic Arg residues, recently
identified shark ghrelin has no amide structure at the C-terminus
of the mature peptide. This is due to the lack of a Gly residue at
the C-terminus of the mature peptide. Goldfish (Unniappan et
al., 2002) ghrelin mRNA has two putative processing sites and
amidation signals after the 12th and 19th amino acids,
suggesting a possible presence of two putative amidated
peptides in this species. However, the endogenous forms of
ghrelin have not been purified.
3.2.2. Mature peptide
Mature ghrelins identified in mammals so far are known to
be a 27- or 28-amino acid peptide (Kojima and Kangawa,
2005). Ghrelin mature peptide is identified in goldfish
(Unniappan et al., 2002), eel (Kaiya et al., 2003a), Mozambique
tilapia (Kaiya et al., 2003b), Nile tilapia (Parhar et al., 2003),
rainbow trout (Kaiya et al., 2003c), channel catfish (Kaiya et al.,
2005), seabream (Yeung et al., 2006), European sea bass
(Terova et al., 2008) and zebrafish (Napolitano, F., 2005
(CAJ20254), referred in Olsson et al., 2008), and has different
lengths from 12- to 23-amino acids (Fig. 2). Recently, ghrelin
was identified from two species of sharks and shark ghrelin, the
first known ghrelin from a cartilaginous fish consists of 26-
amino acids (Kawakoshi et al., 2007). Ghrelin in two species of
frog, Rana catesbeiana (Kaiya et al., 2001) and Rana esculenta
(Napolitano, F., 2005 (AM055941)) are 28-amino acid long.
Turtle ghrelin, the only ghrelin reported so far from reptiles, has
25 amino acids (Kaiya et al., 2004). Ghrelin has been identified
in six species of birds: chicken (Kaiya et al., 2002; Nie et al.,
 Author's personal copy

112
Table 1
Information of ghrelin in non-mammalian vertebrates
Scientific name Ghrelin cDNA accession# Reference Remarks Ghrelin gene accession# Reference GHS-R1a cDNA accession# Reference
Carcharhinus AB254129 Kawakoshi et al. (2007)
melanopterus

H. Kaiya et al. / Comparative Biochemistry and Physiology, Part A 149 (2008) 109–128
Sphyrna lewini AB254128 Kawakoshi et al. (2007) AB254130 Kawakoshi et al. (2007)
Ictalurus punctatus AB196449 Kaiya et al. (2005) Prepro-ghrelin-1
AB196450 Kaiya et al. (2005) Prepro-ghrelin-2
Anguilla japonica AB062427 Kaiya et al. (2003a)
Carassius auratus AF454389 Unniappan et al. (2002) AF454390 Unniappan et al. (2002)
Daino rerio AM055940 Napolitano et al. (2005)⁎ XM_001340372
NM_001083872 XM_001335981 Olsson et al. (2008)
Oreochromis AB077764 Kaiya et al. (2003b) AB361053 Kaiya et al. (2007)⁎
mossambiqus
Oreochromis niloticus AB104859 Parhar et al. (2003) AB104860 Parhar et al. (2003)
Oreochromis urolepis EF371464 Gao et al. (2007)⁎
Epinephelus coioides DQ343147 Lu et al. (2005)⁎
Dicentrarchus labrax DQ665912 Terova et al. (2008)
Acanthopagrus schlegeli AY643808 Yeung et al. (2006) AY643809 Yeung et al. (2006) AY151040 Chan and Cheng (2004)
Oncorhynchus mykiss AB096919 Kaiya et al. (2003c) Prepro-ghrelin-1 AB100839 Kaiya et al. (2003c) AB362479 Kaiya et al. (2007)⁎
AB101443 Kaiya et al. (2003c) Prepro-ghrelin-2
Spheroides nephelus AF082209 Palyha et al. (2000)
Rana catesbeiana AB058510 Kaiya et al. (2001)
Rana esculenta AM055941 Napolitano et al. (2005)⁎
Trachemis scripta AB161457 Kaiya et al. (2002) Prepro-ghrelin-1
elegans
AB161458 Prepro-ghrelin-2

Gallus gallus AB075215 (broiler) Kaiya et al. (2002) AY303688 Nie et al. (2004) AB095995; AJ309542 Tanaka et al. (2003);
Geelissen et al. (2003)
AY299454 (Leghorn) Richards et al. (2006) AB158617 Tanaka et al. (2004)⁎
Anas platyrhynchos AY338466 Yuan et al. (2007)
Dromaius AY338467 Yuan et al. (2007)
novaehollandiae
Anser sp. AY338465 Yuan et al. (2007)
Meleagris gallapavo AY333783 Richards et al. (2006) AY497549 Richards et al. (2006)
Coturnix japonica AB244056 Yoshimura et al. (2006)
Homo sapiens AB029434 Kojima et al. (1999) AF296558 Wajnrajch et al. (2000)
Note: ⁎ These papers have not been published. Database only.
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H. Kaiya et al. / Comparative Biochemistry and Physiology, Part A 149 (2008) 109–128
2004; Richards et al., 2006), turkey (Richards et al., 2006), emu
(Yuan et al., 2007), goose (Yuan et al., 2007), duck (Yuan et al.,
2007) and Japanese quail (Yoshimura et al., 2005; Kaiya et al.,
2007b). Ghrelin in birds is a 26-amino acid peptide. In rats, a
27-amino acid des-Gln14-ghrelin, generated by the alternative
splicing of the ghrelin gene, has been identified (Hosoda
et al., 2000b). This peptide acts as a full agonist of GHS-
R1a. A similar splice variant, des-V13R14Q15-ghrelin was
reported in rainbow trout (Kaiya et al., 2003c). No nucleotide
sequences that cause similar alternative splicing in the region
encoding the mature peptide was found in other non-mammalian
vertebrates.
3.2.3. Acylation at the third amino acid
A unique feature of ghrelin is the modification of Ser-3 with
fatty acid; rat and human ghrelin is modified by octanoic acid
(Kojima et al., 1999; Kojima and Kangawa, 2005). In non-
mammalian vertebrates, almost of all species from cartilaginous
fish to birds have Ser-3, and the amino acid has been modified
mainly by octanoic acid as in mammals. Only frog ghrelin has a
substitution of the amino acid from Ser to threonine (Thr), and
the Thr residue has been modified by octanoic acid (Kaiya et al.,
2001). In fact both Ser-3 and Thr-3 can be modified by various
kinds of middle-chain fatty acids, i.e., decanoic acid (C10:0) and
decenoic acid (C10:1) are attached in human ghrelin (Hosoda
et al., 2003). Similar variations of Ser-3 and Thr-3 modification
have been reported in all non-mammalian vertebrates where the
peptide sequence has been identified (Kaiya et al., 2001, 2003c,
2004, 2005, 2007a; Kawakoshi et al., 2007). In Mozambique
tilapia, ghrelin modified with decanoic acid is the major form of
the mature ghrelin, and octanoylated ghrelin was not detected in
the stomach extracts (Kaiya et al., 2003b).
The exact mechanisms governing acyl modification of
ghrelin remains to be fully elucidated. It is likely that ingested
or absorbed substrates contribute to acylation of ghrelin (Nishi
et al., 2005). It has been reported that oral (OR) or IP
administration of octanoic acid promotes production of acylated
ghrelin in chicken proventriculus (Yamato et al., 2005).
3.2.4. Active core of ghrelin molecule
In mammals, the N-terminal four amino acids of ghrelin
including the acyl modification is essential for receptor binding
and this structure is named as the “active core” of the peptide
(Bednarek et al., 2000; Matsumoto et al., 2001). Comparison of
ghrelin sequences among non-mammalian vertebrates displays
high sequence homology within the seven amino acids at the N-
terminal region (Fig. 2). Goldfish and zebrafish ghrelin have a
substitution in the second amino acid from Ser to Thr
(Unniappan et al., 2002; Kojima and Kangawa, 2005). Bullfrog
ghrelin has two amino acids substituted in the second (Ser to
Leucine (Leu)) and third (Ser to Thr) positions (Kaiya et al.,
2001). On the other hand, there is little identity of the region of
shark ghrelins (GVSFHPR), although Ser-3 has been modified
by octanoic or decanoic acid (Kawakoshi et al., 2007).
Interestingly, the shark ghrelin can increase intracellular
calcium concentration of the cells expressing rat GHS-R1a,
indicating the peptide can bind to rat GHS-R1a.
113
3.2.5. Des-acyl ghrelin
Hosoda et al. (2000a) reported presence of two forms of
ghrelin in gastrointestinal tissues of rats, ghrelin having
acylation at Ser-3 and des-acyl ghrelin lacking the acyl
modification at Ser-3. Des-acyl ghrelin is also present in
circulation and growing evidence suggests des-acyl ghrelin has
some biological actions (Asakawa et al., 2005; Chen et al.,
2005; Toshinai et al., 2006; Sato et al., 2006). In non-
mammalian vertebrates, Kaiya et al. (2006a) revealed presence
of des-acyl ghrelin-like substance in the stomach and plasma of
bullfrog using radioimmunoassay (RIA) combined with high-
performance liquid chromatography (HPLC). Furthermore, des-
acyl ghrelin can inhibit ghrelin-induced food intake in goldfish,
although des-acyl ghrelin alone has no effect (Matsuda et al.,
2006a). These results suggest synthesis and bioactivity of des-
acyl ghrelin in non-mammalian vertebrates.
3.2.6. Obestatin
Zhang et al. (2005) discovered a peptide that antagonizes the
stimulatory effect of ghrelin on food intake and gut motility
though an orphan GPCR, GPR39. The peptide was found in the
C-terminal region of rat prepro-ghrelin. This peptide was named
obestatin, a contraction of obese from Latin “obedere”, meaning
to devour, and “statin” denoting suppression. Obestatin is a 23-
amino acid amidated peptide, and putative obestatin 13–23 has
also been present. However, several groups who followed up
these studies were unable to reproduce the original findings on
obestatin binding and activation of GPR39 in vitro (Lauwers et
al., 2006; Holst et al., 2007; Chartrel et al., 2007). Zhang et al.
(2007) confirmed impurities in the initial batch of obestatin they
used, and indicated that they could not reproduce their original
finding on obestatin binding and activation of GPR39. Zhang et
al. (2007) claims reproducibility of inhibitory effects of
obestatin on food intake. However, several other researchers
were unable to find any inhibitory effect for obestatin on food
intake and body weight gain (Bresciani et al., 2006; Seoane et
al., 2006; Sibilia et al., 2006; Zizzari et al., 2007). Nothing is
reported about the isolation of obestatin in non-mammalian
vertebrates, although possible existence has been proposed in
seabream (Yeung et al., 2006). However, the obestatin region in
non-mammalian prepro-ghrelin sequences is highly divergent
compared to mammalian obestatins (Fig. 2). It was proposed to
obestatin as ghrelin-associated peptide (Gourcerol and Taché,
2007; Gourcerol et al., 2007). Given that the “obestatin”
sequence does not exist in non-mammalian vertebrates and its
synthesis from proghrelin in mammals is not clear, re-naming
this sequence as ghrelin-associated peptide could lead to further
confusions in this already controversial research field.
4. Tissue distribution and localization of ghrelin
In rats, ghrelin is primarily produced in the mucosal layer of
stomach from the classically known X/A-like cells that is now
known as ghrelin cells (Kojima et al., 1999; Date et al., 2000).
Ghrelin immunoreactive cells are also found in the duodenum,
and their number are decreased gradually in distal intestines from
ileum to colon (Date et al., 2000). Ghrelin is also produced in the
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H. Kaiya et al. / Comparative Biochemistry and Physiology, Part A 149 (2008) 109–128 115

Table 2
Tissue distribution of ghrelin in vertebrates
Hammerhead shark Eel Goldfish Catfish Rainbow trout Tilapia Bullfrog Turtle Chicken Rat
Brain ++ ++ +++ N.D. ++ +++ – – ++ +
Hypothalamus ++ N.D. + + ++ N.D. N.D. N.D. N.D. +
Pituitary N.D. N.D. – ++ N.D. N.D. N.D. N.D. N.D. +
Heart +++ + – + +++ – – – – +
Lung N.P. N.P. N.P. N.P. N.P. N.P. + – + +
Stomach ++++ ++++ N.P. ++++ ++++ ++++ ++++ ++++ ++++ ++++
Pancreas N.D. N.D. N.D. +++ N.D. N.D. + + – +
Spleen + N.D. ++++ + + N.D. – – + N.D.
Liver ++ N.D. + + – N.D. – – – +
Intestine +++ ++++ +++ ++ +++ – ++ ++ + +++
Rectal gland + N.P. N.P. N.P. N.P. N.P. N.P. N.P. N.P. N.P.
Adrenal gland N.P. N.P. N.P. N.P. N.P. N.P. – – N.D. +
Head kidney N.D. + N.D. + + N.D. N.P. N.P. N.P. N.P.
Trunk kidney + ++ – ++ + – – – – +
Gall bladder N.D. N.D. N.D. +++ N.D. N.D. + N.D. N.D. N.D.
Ovary N.D. N.D. – N.D. N.D. N.D. N.D. N.D. + N.D.
Testis N.D. N.D. – N.D. N.D. N.D. + N.D. N.D. +
Gill ++ – +++ ++ + + N.D. N.P. N.P. N.P.
This table showed relative level of ghrelin based on expression level of ghrelin mRNA or ghrelin content.
++++: very high; +++: high; ++: moderate; +: low; –: not detected; N.D.: not determined; N.P.: not present.

hypothalamic arcuate nucleus (Kojima et al., 1999; Lu et al.,
2002; Mondal et al., 2005), pancreatic A-cells (Kageyama et al.,
2005), kidney (Mori et al., 2000) and pituitary (Korbonits et al.,
2001) of rats, mice and humans. However, the expression is
relatively lower than that of the gastrointestinal tract (Hosoda et
al., 2000a,b).
In non-mammalian vertebrates, ghrelin peptide was purified
from stomach extracts, and ghrelin immunoreactivity and its
mRNA expression have been detected mainly in the stomach or
intestine, indicating that gastrointestinal tract is the major site of
ghrelin production in vertebrates. However, expression of
ghrelin mRNA was found in various organs of non-mammalian
vertebrates other than stomach and there is a species-specific
difference in ghrelin expression. For example, ghrelin expres-
sion in the brain was found in chicken, eel, Mozambique tilapia,
rainbow trout, catfish and hammerhead shark, but not in turtle.
Expression in the intestine was found in chicken, turtle, eel,
rainbow trout, catfish and hammerhead shark, but not in
Mozambique tilapia. Ghrelin mRNA is expressed in the kidney
of eel, rainbow trout and catfish, but not in chicken, turtle,
Mozambique tilapia and hammerhead shark. Table 2 sum-
marizes the tissue distribution of ghrelin in vertebrates. The
following sections will highlight our knowledge of ghrelin
distribution in the stomach and brain, and ghrelin levels in blood
and tissues in non-mammals.
4.1. Stomach
Ghrelin mRNA expression is found in the stomach of
bullfrog and chicken by Northern blot analysis (Kaiya et al.,
2001, 2002). Ghrelin mRNA expression using reverse-tran-
scription-polymerase chain reaction (RT-PCR) was detected in
the stomach of chicken (Kaiya et al., 2002; Wada et al., 2003),
quail (Yoshimura et al., 2005), bullfrog (Kaiya et al., 2001),
turtle (Kaiya et al., 2004), Japanese eel (Kaiya et al., 2003a),
tilapia (Parhar et al., 2003; Kaiya et al., 2003b), rainbow trout
(Kaiya et al., 2003c), channel catfish (Kaiya et al., 2005),
seabream (Yeung et al., 2006), sea bass (Terova et al., 2008) and
hammerhead shark (Kawakoshi et al., 2007). Goldfish lacks a
stomach, but northern blot and RT-PCR analyses demonstrated
a strong expression of ghrelin mRNA in the gut (Unniappan
et al., 2002). Therefore, the gastrointestinal tract is recognized
as the main site for ghrelin production in non-mammalian
vertebrates.
Ghrelin-immunoreactive (ir) cells are detected in the
mucosal layer of the stomach in chicken (Wada et al., 2003;
Yamato et al., 2005), quail (Yoshimura et al., 2005), turtle
(Kaiya et al., 2004), frogs (Galas et al., 2002; Kaiya et al.,
2007b), rainbow trout (Sakata et al., 2004) and Japanese eel
(Kaiya et al., 2006b). Recently, Olsson et al. (2008) reported
localization of ghrelin-ir cells in the gut of zebrafish. The
antibody used in these studies recognizes only acylated (active-
form) ghrelin, indicating that immunopositive cells show the
presence of bioactive ghrelin (Hosoda et al., 2000a). Also
recently, Kaiya et al. (2006a) showed localization of ghrelin-ir
cells in the bullfrog stomach using the antisera raised against
bullfrog ghrelin C-terminal fragment 13–28. This antibody can
recognize both acylated and des-acyl ghrelin, and is character-
istically similar to the antisera for C-RIA (Hosoda et al., 2000a).
In chicken, Neglia et al. (2005) found ghrelin-ir cells in the

Fig. 2. Comparison of prepro-ghrelin peptides from various non-mammalian vertebrates with human prepro-ghrelin sequence. Signal peptide, mature peptide and C-
terminal peptide (C-peptide) regions of the prepro-ghrelin are shown separately. Number of amino acids in amidated mature ghrelin is provided in parentheses of the
middle panel. Black-boxed letter indicates a glycine residue that contributes to create an amide structure of mature ghrelin in teleosts. Parentheses in the lower panel for
C-peptide shows total number of amino acids for prepro-ghrelin with exceptions of edible frog and quail for incomplete sequence. Obestatin region in the C-peptide of
human sequence is shown with broken lines. Prepro-ghrelin sequences were obtained from publications reporting its cloning or from the GenBank (accession numbers
provided in Table 1). Sequences were aligned using the GENETYX-Mac software program.
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116 H. Kaiya et al. / Comparative Biochemistry and Physiology, Part A 149 (2008) 109–128
gastrointestinal tissues, but Ahmed and Harvey (2002) failed to
detect ghrelin-ir cells in the proventriculus using commercially
available antibodies. The difference is due to the variation of
recognition site of the antibodies to ghrelin molecule.
4.2. Brain
Using RT-PCR, ghrelin mRNA expression is detected in the
brain of chicken (Kaiya et al., 2002; Saito et al., 2005; Richards
et al., 2006; Kaiya et al., 2007b), goldfish (Unniappan et al.,
2002), Mozambique tilapia (Kaiya et al., 2003b), rainbow trout
(Kaiya et al., 2003c), Japanese eel (Kaiya et al., 2003a) and
hammerhead shark (Kawakoshi et al., 2007). In chicken, various
parts of brain, including corpus striatum, cerebellum, optic lobe
and brainstem, express ghrelin mRNA, and the corpus striatum
showed the highest expression. In goldfish, telencephalon,
followed by hypothalamus, express ghrelin mRNA. In hammer-
head shark, dorsal nucleus, followed by forebrain, cerebellum,
hypothalamus and optic tectum were expression sites for the
ghrelin gene. On the other hand, no ghrelin mRNA expression
was detected in the brain of the bullfrog (Kaiya et al., 2001),
turtle (Kaiya et al., 2004), Nile tilapia (Parhar et al., 2003),
catfish (Kaiya et al., 2005), seabream (Yeung et al., 2006) and
sea bass (Terova et al., 2008). Further studies are necessary to
validate the presence of ghrelin mRNA in specific brain regions
of these animals.
In the mammalian brain, ghrelin-ir cells are localized in
arcuate nucleus of the hypothalamus, as demonstrated in rats
(Kojima et al., 1999). In addition, Cowley et al. (2003) reported
a wide distribution of ghrelin-ir neurons and fibers in rats.
Among non-mammalian vertebrates, the detailed distribution of
ir-ghrelin in the brain has been reported only in frogs and
chicken. Ghrelin-ir neurons were found in the diencephalon and
mesencephalon of the frog, R. esculenta (Galas et al., 2002). In
addition, a wide distribution of ghrelin-ir nerve fibers was also
detected in the telencephalon, diencephalon and mesencephalon
(Galas et al., 2002). In chicken, ghrelin-ir cells and nerve fibers
are observed in the hypothalamus (Ahmed and Harvey, 2002).
However, as the rat ghrelin antibody did not detect ir-ghrelin in
chicken proventriculus, it has been suggested that the antibody
cross-reacts with a protein other than ghrelin in the chicken
hypothalamus (Wada et al., 2003). No staining for ghrelin was
found in the pituitary and brain of a broiler strain using the
antibody for the active-form of ghrelin (S. Geelissen, R. Wada
and T. Sakai, personal communications).
4.3. Ghrelin levels in plasma or tissues
Hosoda et al. (2000a) validated two RIAs for ghrelin
measurement in mammals; the N-RIA that can measure
acylated, active-form of ghrelin and the C-RIA, that can
measure the total concentration of ghrelin including acylated
and unacylated des-acyl ghrelin. Using the N-RIA, acylated
ghrelin concentrations in rodent plasma are detected from
approximately 5 to 80 fmol/mL (Hosoda et al., 2000a; Ariyasu
et al., 2005), and acylated ghrelin content in the rat stomach
ranges from 370 to 1800 fmol/mg tissue (Hosoda et al., 2000a;
Date et al., 2000). Total ghrelin levels in plasma ranged from
150 to 640 fmol/mL in rats, mice and humans (Hosoda et al.,
2000a; Date et al., 2000; Yoshimoto et al., 2002; Ariyasu et al.,
2002; Akamizu et al., 2004). Total ghrelin levels in the stomach
of rats range from 1800 to 4600 fmol/mg tissue (Hosoda et al.,
2000a,b; Date et al., 2000).
In non-mammalian vertebrates, Mustonen et al. (2002)
reported plasma levels of ghrelin in a fish, the burbot (Lota lota)
before spawn as approximately 470 pg/mL (140 fmol) using a
commercial RIA kit for human ghrelin. They reported the levels
reduced during spawn to 350 pg/mL and increased to 780 pg/
mL after spawn. Nieminen et al. (2003) reported similar levels
of plasma ir-ghrelin in burbot (400 pg/mL) and the levels
reduced by two-week fasting at 2 °C. These results suggest that
ghrelin may be involved in energy homeostasis in burbot. In
goldfish, serum ghrelin levels were found to be 50 fmol/mL at
normal feeding state, using a validated RIA that detected the
active-form of ghrelin (Unniappan et al., 2004). Serum ghrelin
levels significantly increased 3 days after fasting in goldfish,
and reached peak levels at 5 days. A post-prandial decrease in
the serum ghrelin concentration was also observed. These
results indicate that ghrelin participates in feeding regulation in
goldfish. Kaiya et al. (2006b) validated the N-RIA for eel
ghrelin; plasma ghrelin levels were approximately 40 fmol/mL
in freshwater (FW)-adapted Japanese eels starved for 1 week,
and the levels transiently increased to 200 fmol/mL at 6 h after
transfer from FW to seawater (SW), suggesting a possible
osmoregulatory action of ghrelin in eels. Acylated ghrelin
content in the eel stomach was approximately 40 fmol/mg wet
tissue, and the levels did not change after transfer of eels from
FW to SW (Kaiya et al., 2006b). In rainbow trout, active ghrelin
levels in plasma have been reported as 4–8 fmol/mL (Jönsson et
al., 2007), and no pre-prandial and post-prandial changes in
plasma ghrelin levels were observed.
In amphibians, Kaiya et al. (2006a) established a RIA for the
measurement of total ghrelin concentration of bullfrog. Ghrelin
levels in plasma and stomach extracts were approximately 50–
150 fmol/mL and 83–135 fmol/mg wet tissue, respectively. In
bullfrog plasma, the majority of ghrelin immunoreactivity was
des-acyl ghrelin and C-terminal fragments of ghrelin. On the
other hand, the major form of ghrelin stored in the bullfrog
stomach was acylated ghrelin. Total ghrelin levels in plasma and
stomach significantly increases in frogs fasted for 10 days and
for 20 days, suggesting a possible involvement of ghrelin in
feeding behavior in bullfrog.
In birds, Kaiya et al. (2007b) have reported that active
ghrelin concentrations in chicken plasma are approximately
20 fmol/mL using the N-RIA for chicken ghrelin, and the levels
increase to about 30 fmol/mL 12 h after fasting. The increased
ghrelin levels returned to the control levels 6-h after re-feeding.
These results suggest that ghrelin plays a role in “hunger signal”
in chicken.
5. Ghrelin receptor (GHS-R)
As mentioned earlier, GHS-R is a G-protein-coupled
receptor (GPCR) firstly identified in pig and human (Howard
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H. Kaiya et al. / Comparative Biochemistry and Physiology, Part A 149 (2008) 109–128 117

et al., 1996). Two types of GHS-R, GHS-R1a that mediates GHS-R1a-expressing mammalian cells (Howard et al., 1996;
biological actions of GHS and ghrelin, and an inactive, Kojima et al., 1999) or in rat and human pituitary cells
alternative splice variant GHS-R1b have been known. In (Herrington and Hille, 1994; Lania et al., 1998), biphasic Ca2+

Fig. 3. Multiple sequence alignment of the ghrelin receptor (GHS-R) 1a protein in human, chicken, pufferfish, seabream, zebrafish, tilapia and rainbow trout.
Accession number is human GHS-R1a (AY429112), chicken GHS-R1a (AB095995; AJ309542), pufferfish 78B7 (AF082209), seabream GHS-R1a (AY151040),
zebrafish GHS-R1a (XM001335981), zebrafish GHS-R2a (XM001340372), tilapia GHS-R1a (AB361053), rainbow trout GHS-R1a (AB362479).
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118 H. Kaiya et al. / Comparative Biochemistry and Physiology, Part A 149 (2008) 109–128

Fig. 4. Multiple sequence alignment of the ghrelin receptor (GHS-R) 1b protein in human, chicken, seabream, tilapia and rainbow trout. Accession number is human
GHS-R1b (U60181), chicken GHS-R1aV (AB095996), seabream GHS-R1b (AY151041), tilapia GHS-R1b (AB361054) and rainbow trout GHS-R1b (AB362481).

increases, which due to a transient Ca2+ release from the
intracellular store and Ca2+ influx through voltage-dependent
L-type calcium channel, are observed as the signal transduction.
Among non-mammalian vertebrates, GHS-R has been
reported in two species of teleosts, black seabream and a
pufferfish, and in a bird, chicken (Figs. 3 and 4). In the black
seabream, both types of receptor, GHS-R1a (385 amino acids
(aa)) and GHS-R1b (295 aa), have been identified (Chan and
Cheng, 2004; Chan et al., 2004b). In a pufferfish, Spheroides
nephelus, GHS-R1a ortholog, namely 78B7 (374 aa) has been
reported (Palyha et al., 2000). In chicken, GHS-R1a (347 aa)
(Geelissen et al., 2003; Tanaka et al., 2003), GHS-R1aV (331
aa) (Tanaka et al., 2003) and GHS-R1c (Geelissen et al., 2003)
have been reported. Amino acid sequences of GHS-R1a of
seabream, pufferfish and chicken GHS-R1a have 56, 55 and
72% identity to human GHS-R1a, respectively (Fig. 3). GHS-
R1aV and GHS-R1c lack the same portion of 48 bp in the
transmembrane domain-6. Here we propose that nomenclature
of these two receptors be standardized as GHS-R1aV since
these receptor proteins are identical, and since it is a partial
truncated form of GHS-R1a. In addition, Sirotkin et al. (2006)
reported the possible presence of a third GHS-R variant
identified in chicken ovary, namely cGHS-R1tv. Unfortunately,
the nucleotide and amino acid sequences of the cGHS-R1tv is
unavailable. It is likely that this receptor may be an ortholog
of GHS-R1b as seen in other animals with judging from its
generation process. Recently, GHS-R has been identified in
zebrafish (Olsson et al., 2008), Mozambique tilapia and rain-
bow trout (unpublished observations by H. Kaiya). In zebrafish,
Olsson et al. (2008) reported 360-amino acid protein. On the
other hand, we discovered another GHS-R paralogous sequence
using commercial zebrafish genomic database, which has 365-
amino acid and shows 74% homology with the a 360-amino
acid zebrafish GHS-R at amino acid level. Therefore, we
propose that nomenclature of the firstly identified 360-amino
acid protein and the second 365-amino acid protein are named
as zebrafish GHS-R1a and GHS-R2a, respectively (Fig. 3).
Whether these receptors are functional remain unknown. In
Mozambique tilapia, both GHS-R1a (384 aa) and GHS-R1b
(298 aa) have been identified (Kaiya et al., 2007b, database
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H. Kaiya et al. / Comparative Biochemistry and Physiology, Part A 149 (2008) 109–128
only). Rainbow trout GHS-R1a and 1b were composed of 387
aa and 300 aa, respectively (Kaiya et al., 2007b, database only).
Amino acid sequences of GHS-R1a of Mozambique tilapia and
rainbow trout have 55 and 57% identity to human GHS-R1a,
respectively.
It has been reported that the GHS-R gene has two exons and
one intron in chicken (Tanaka et al., 2003), seabream (Chan and
Cheng, 2004) and pufferfish (Palyha et al., 2000). Recently
identified tilapia and rainbow trout GHS-R genes have also
composed of two exons and one intron (Kaiya et al., 2007b,
database only). This gene organization and the insert position of
the intron are identical to those of the mammalian GHS-R gene
(McKee et al., 1997; Petersenn, 2002), indicating that the GHS-
R gene has been evolutionary conserved from fish to mammals.
Yeung et al. (2004) isolated 5′-flanking region of the seabream
GHS-R gene, and reported presence of a number of unique
putative transcription factor-binding sites different from human
counterpart. Interestingly, a GHS-R agonist, GHS enhances the
promoter activity, and the stimulatory effect is blocked by a
GHS-R antagonist, indicating a homologous up-regulation of
transcription of the GHS-R gene in seabream. This is opposite
to the regulation in human GHS-R, where the promoter activity
is inhibited by the treatment with a GHS-R agonist.
GHS-R1a mRNA is mainly expressed in the pituitary and
various parts of the brain of chicken (Geelissen et al., 2003;
Tanaka et al., 2003; Saito et al., 2005; Richards et al., 2006) and
seabream (Chan and Cheng, 2004), suggesting direct actions of
ghrelin in these tissues. The GHS-R splice variants (GHS-R1b,
1aVor 1c) are detected in almost all tissues that expressed GHS-
R1a (Geelissen et al., 2003; Tanaka et al., 2003; Chan and
Cheng, 2004). However, the expression level of GHS-R splice
variant mRNAs is much lower than that of the GHS-R1a.
Furthermore, the physiological function of GHS-R1b remains to
be fully elucidated, although it has been reported that the
seabream GHS-R1a activity is attenuated to some extent when
co-expressed with seabream GHS-R1b in HEK293 cells (Chan
and Cheng, 2004). Expression of GHS-R1a and 1b mRNA were
detected in the phagocytic leukocytes of rainbow trout, and
GHS-R1a is involved in immune functions of the leukocytes
through production of local GH by ghrelin (Yada et al., 2006).
The signal transduction mechanism of GHS-R in non-
mammalian vertebrates has been investigated using mammalian
HEK293 cells expressing seabream GHS-R1a (Chan et al.,
2004b). They concluded that seabream GHS-R1a is a
constitutively active receptor, which triggers a PLC-PKC-
dependent [Ca2+] mobilization in association with L-type
voltage-gated calcium channel by PKC. Thus, it is likely that
the characteristics of GHS-R on signal transduction in the cells
are similar among seabream, rat and human (Herrington and
Hille, 1994; Lania et al., 1998; Holst et al., 2005).
6. Biological actions of ghrelin
In mammals, ghrelin participates in several physiological
functions other than stimulation of GH release. Several reviews
have discussed the details (van der Lely et al., 2004; Kojima and
Kangawa, 2005; Peeters, 2005; Hosoda et al., 2006). In non-
119
mammalian vertebrates, ghrelin is also involved in various
physiological functions (Table 3). Unniappan and Peter (2005),
and Kaiya et al. (2007b) have previously summarized ghrelin
function in fish and birds, respectively.
6.1. Regulation of hormone release and synthesis
A variety of evidence indicates that ghrelin directly acts on
the pituitary in fish, amphibians, and birds. In addition, it is
likely that circulating ghrelin directly or indirectly acts as an
endocrine peptide to stimulate the release of some hormones,
although the regulatory mechanisms need further investigation.
6.1.1. Fish
Ghrelin directly affects on the pituitary of fish. In organ-
cultured pituitary of Mozambique tilapia, the release of GH and
prolactin (PRL) is simulated by ghrelin originated from rat, eel,
and tilapia (Riley et al., 2002; Kaiya et al., 2003a,b). Recently,
Fox et al. (2007) reported that tilapia ghrelin-stimulated GH
release, but not PRL release from dispersed pituitary cells of the
tilapia. This suggests a possible importance of cell-to-cell
communication for regulation of PRL release by ghrelin. In the
study, it has been shown that an effect of tilapia ghrelin C-10
was more potent than that of tilapia ghrelin C-8. In addition, a
GHS-R antagonist, [D-Lys3]-GHRP-6 blocked the ghrelin's
effects, indicating that ghrelin's effects are mediated by GHS-
R1a on the pituitary cells. In seabream, GHS stimulates GH
release from the pituitary (Chan et al., 2004a). The direct action
of ghrelin on the pituitary are supported by the fact that GHS-
R1a is present in seabream pituitary (Chan and Cheng, 2004). In
goldfish, synthetic goldfish ghrelin stimulates the release of GH
from perifused pituitary cells and pituitary cells in static culture
(Unniappan and Peter, 2004). Ran et al. (2004) also observed a
GH-releasing activity from pituitary fragments by rat ghrelin in
orange-spotted grouper. In rainbow trout, trout ghrelin stimu-
lated the release of only GH, but not PRL and somatolactin (SL)
in organ-cultured pituitary (Kaiya et al., 2003c). Furthermore,
ghrelin-stimulated GH release from goldfish pituitary cells is
inhibited by somatostatin-14 (Unniappan and Peter, 2004),
suggesting a similar intracellular linkage of ghrelin with
classical regulation of GH secretion.
Ghrelin also affects on pituitary hormone secretion through
circulation. Circulating levels of GH increase after IP injections
of a GHS, KP-102 in Nile tilapia (Shepherd et al., 2000), after
ICV and IP injections of ghrelin in goldfish (Unniappan and
Peter, 2004), after IP injection of ghrelin in channel catfish
(Kaiya et al., 2005) and rainbow trout (Kaiya et al., 2003c;
Shepherd et al., 2007), and after IP injections of tilapia ghrelin
in Mozambique tilapia (Fox et al., 2007). Chronic administra-
tion of ghrelin using osmotic mini-pumps, however, caused no
change in circulating GH levels after 21 days post-implantation
(Riley et al., 2005). Similar findings have been reported in rats
(Bowers et al., 1984; Date et al., 2000). It may be due to
desensitization of the GH secretory response of somatotrophs
during ghrelin administration. Other than GH, no changes in
circulating levels of PRL are observed after IP injection of a
GHS in Nile tilapia, of trout ghrelin in rainbow trout or of tilapia
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120 H. Kaiya et al. / Comparative Biochemistry and Physiology, Part A 149 (2008) 109–128

Table 3 Summary of ghrelin function examined in non-mammalian vertebrates

Cartilaginous fish Teleostian Amphibian Reptilian Avian Mammalian
Hormone secretion
GH N.D. ↑ ↑ N.D. ↑ ↑
PRL N.D. ↑→ ↑ N.D. N.D. ↑(weak)
SL – → – – – –
ACTH N.D. N.D. N.D. N.D. (↑) ↑(weak)
FSH N.D. – → N.D. → →
LH N.D. ↑ → N.D. N.D. ↑? →
TSH N.D. N.D. N.D. N.D. → ↓? →
TRH N.D. N.D. N.D. N.D. ↑→ →
CRF N.D. N.D. N.D. N.D. ↑→ ↑?↓
Corticosteroids N.D. N.D. N.D. N.D. ↑ ↑(weak)
Insulin N.D. N.D. N.D. N.D. N.D. ↑?↓
Behaviour
Food intake N.D. ↑→ (↑) N.D. ↓ ↑
Water intake N.D. ↓ N.D. N.D. ↓ ↓
Locomotor activity N.D. ↑ N.D. N.D. ↑↓ ↑↓
Metabolism
Adiposity N.D. ↑ N.D. N.D. N.D. ↑
Blood glucose N.D. N.D. N.D. N.D. N.D. ↑?↓
Gastrointestinal function
Acid secretion N.D. N.D. N.D. N.D. N.D. ↑
Motility N.D. N.D. N.D. N.D. ↑ ↑
Turnover of mucosa N.D. N.D. N.D. N.D. N.D. ↑
Cardiovascular function
Cardiac output N.D. N.D. N.D. N.D. N.D. ↑
Blood pressure N.D. N.D. N.D. N.D. N.D. ↓
Note: N.D.: not determined; ↑: increase; ↓: decrease; →: no change.

ghrelin in Mozambique tilapia (Shepherd et al., 2000; Kaiya et
al., 2003c; Fox et al., 2007). As mentioned before, ghrelin can
stimulate PRL release of Mozambique tilapia from organ-
cultured pituitary (Riley et al., 2002; Kaiya et al., 2003a,b).
Further studies are necessary to identify the reasons for different
PRL response in tilapia and rainbow trout. Besides, plasma SL
levels did not change after IP injections of trout ghrelin in
rainbow trout (Kaiya et al., 2003c). Unniappan and Peter (2004)
have reported a small increase in serum LH levels in goldfish
after ICV or IP injection of goldfish ghrelin. In rainbow trout,
Shepherd et al. (2007) observed increases in plasma IGF-I
levels 3 h after IP injection of KP-102 or rat ghrelin. Similarly,
Fox et al. (2007) reported IGF-I increase in tilapia plasma 10 h
after IP injections of tilapia ghrelin. Ghrelin affects gene
expression of pituitary hormones. Ghrelin increased GH mRNA
expression in the pituitary of orange-spotted grouper (Ran et al.,
2004) and goldfish (Unniappan and Peter, 2004) in vitro,
suggesting a role for ghrelin in regulating GH synthesis in fish.
IP injections of ghrelin also increased GH mRNA expression in
the pituitary of channel catfish and Mozambique tilapia (Kaiya
et al., 2005; Fox et al., 2007). Riley et al. (2005) reported a
different stimulatory action on GH synthesis in Mozambique
tilapia with tilapia ghrelin C-8 and C-10; ghrelin C-8 increased
GH mRNA but ghrelin C-10 did not. These in vitro and in vivo
results indicate that ghrelin stimulates not only the release but
also the synthesis of GH in some fish. In goldfish, LH mRNA
expression, as well as LH secretion, is stimulated by ghrelin
(Unniappan and Peter, 2004).
IP injections of ghrelin were found to cause a significant
reduction in prepro-somatostatin-II mRNA expression in the
goldfish brain (Canosa et al., 2005), indicating possible
interactions between ghrelin and somatostatin in the regulation
of GH release in goldfish. Hepatic IGF-I mRNA increased by IP
injections of tilapia ghrelin in Mozambique tilapia (Fox et al.,
2007), suggesting that the GH-IGF-I axis is mediated by
ghrelin. Recent studies suggest the possible interactions of
ghrelin with other peptides such as gastrin-releasing peptide/
bombesin (GRP/BBS) to regulate the post-prandial GH surge in
goldfish (Canosa et al., 2005). Further studies are necessary to
elucidate the actions and interactions of ghrelin with other
releasing and release-inhibitory factors in regulating GH
secretion and release of other pituitary hormones in fish.
6.1.2. Amphibians
Bullfrog and rat ghrelin stimulate the release of GH and PRL
from dispersed pituitary cells of the bullfrog without any effects
on LH and FSH release (Kaiya et al., 2001). This result indicates
that ghrelin directly acts on the somatotrophs and lactotrophs of
bullfrog. Interestingly, the potency is 100–1000 times greater
for bullfrog ghrelin than for rat ghrelin, which may be due to the
differences in the structure and affinity of these peptides to the
bullfrog ghrelin receptor. Bullfrog ghrelin increases plasma GH
and PRL levels 15 min after injection when injected IP into
juvenile bullfrogs and the elevated levels returns to the basal
levels by 60 min after the injection (H. Kaiya, A. Koda, K.
Yamamoto, S. Kikuyama, unpublished data). This suggests a
direct action of circulating ghrelin on somatotrophs and
lactotrophs and the possible existence of a signal transduction
with the vagal system in the bullfrog, as demonstrated in rats
(Date et al., 2002).
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H. Kaiya et al. / Comparative Biochemistry and Physiology, Part A 149 (2008) 109–128
6.1.3. Birds
Ghrelin stimulates GH release in vivo and in vitro in chicken
(Ahmed and Harvey, 2002; Kaiya et al., 2002; Baudet and
Harvey, 2003). Ahmed and Harvey (2002) observed an increase
in plasma GH levels 10 min after IV injection of rat ghrelin in
chicken. The effect was comparable with that of GHRH, but
much weaker than that of TRH. Kaiya et al. (2002) observed the
same effects using a homologous chicken ghrelin. In this study,
plasma GH levels transiently increased 15 min after IV
injection. In addition to the GH increase, plasma corticosterone
levels also increased and the dose–response relationship was
much clearer in corticosterone than GH (Kaiya et al., 2002).
ICV injections of chicken ghrelin also increase plasma
corticosterone levels (Saito et al., 2005). Thus, ghrelin would
act on pituitary or adrenal gland directly or indirectly through
the hypothalamo-pituitary-adrenal axis (HPA-axis). Geelissen
et al. (2006) reported no effects for chicken ghrelin on thyroxine
(T3) levels in chicken after IV injections, suggesting no effect
for ghrelin on at least TSH release from the pituitary.
6.1.4. Reptiles
There are no reports currently available on the in vitro or in
vivo effects of ghrelin in the regulation of any hormone release
in reptiles. From a phylogenic point of view, it is valuable to
investigate the role of ghrelin in the regulation of hormone
release in reptiles.
6.2. Appetite regulatory effects
Ghrelin is an orexigen in mammals (Nakazato et al., 2001;
Tschöp et al., 2000; Wren et al., 2000, 2001). Ghrelin stimulates
food intake when injected centrally or peripherally, and it is
known that various hypothalamic neurons such as NPY/AGRP,
orexin, and POMC interact with the ghrelin signal system
(Horvath et al., 2001; Date et al., 2006; Gil-Campos et al., 2006).
A variety of evidence supports the participation of ghrelin in
modifying feeding behaviors in non-mammalian vertebrates.
6.2.1. Fish
The regulation of food intake has been studied using
goldfish, tilapia and rainbow trout. Central administration of
human ghrelin, octanoylated 19-amino acid and 12-amino acid
goldfish ghrelins stimulate food intake in goldfish (Unniappan
et al., 2002; Unniappan et al., 2004). Matsuda et al. (2006a,b)
also found that ICV injections of octanoylated, 12-amino acid
goldfish ghrelin stimulate food intake in goldfish. Recently,
Miura et al. revealed that ICV ghrelin-induced food intake in
goldfish is mediated by NPY and orexin, as seen in rodents
(Miura et al., 2006, 2007).
IP injections of ghrelin are also effective to stimulate food
intake (Unniappan et al., 2004; Matsuda et al., 2006a,b). The
peripheral ghrelin-induced food intake in goldfish is mediated
by a mechanism through capsaicin-sensitive vagal afferent, as
demonstrated in rats (Date et al., 2002; Matsuda et al., 2006a).
In agreement with these studies using acute injections of
ghrelin, chronic administration of ghrelin using osmotic mini-
pumps for 21 days also resulted in an increase in food intake and
121
weight gain of the Mozambique tilapia (Riley et al., 2005). It is
interesting to note that the effect has been observed with
decanoylated tilapia ghrelin C-10, which is the major form in
the tilapia, but not with octanoylated tilapia ghrelin C-8.
Shepherd et al. (2007) observed increased food intake 2 or 5 h
after IP injections of several GH secretagogues such as KP-102
and rat ghrelin in juvenile rainbow trout. On the other hand,
Jönsson et al. (2007) has observed no effect of trout ghrelin on
food intake in two-year-old rainbow trout during 12-h post-IP
injections of trout ghrelin. These results may suggest a species-
specific and a developmental stage-specific action of ghrelin on
feeding regulation in fish.
It has been reported that ghrelin mRNA expression and
ghrelin secretion changes during feeding conditions. A post-
prandial decrease in ghrelin mRNA expression in the
hypothalamus and gut, and a post-prandial decrease in serum
ghrelin levels are observed in goldfish (Unniappan et al., 2004;
Canosa et al., 2005). Fasting for 7 days caused a significant
increase in ghrelin mRNA expression in the hypothalamus and
gut of goldfish (Unniappan et al., 2004). A significant increase
in serum ghrelin levels is observed 3 days after fasting, the
increase reaches the maximum at 5 days, and a little decrease
in serum ghrelin is observed at 7 days after fasting. Matsuda et
al. (2006b) also observed a significant increase in ghrelin
mRNA expression in the gut, but such an increase was not
observed in the brain. This may be due to the very low
expression of the ghrelin gene in the hypothalamus. These
meal-related increases in ghrelin mRNA expression in the
brain and gut, and ghrelin levels in blood support the view that
ghrelin is involved in the regulation of food intake in goldfish.
In burbot (Lota lota), however, a significant reduction in
plasma ir-ghrelin levels was observed after a two-week fast at
2 °C, but not at 10 °C (Nieminen et al., 2003). This is an
opposite response compared with other animals. Furthermore,
in Nile tilapia, it has been demonstrated using quantitative RT-
PCR that ghrelin mRNA levels remain unchanged after 7-day
food deprivation (Parhar et al., 2003). Food intake in fish is
regulated by the co-ordination of multiple, redundant neu-
roendocrine pathways (Volkoff et al., 2005). The lack of an up-
regulation in ghrelin mRNA expression in Nile tilapia after
7 days of starvation might be due to a compensatory decrease
or increase in one or more of the appetite regulatory peptides.
IP injections of the anorexigen GRP/BBS inhibit ghrelin
mRNA expression in the gut of goldfish (Canosa et al., 2005),
suggesting interactions between ghrelin and other appetite
regulatory peptides in fish.
Des-acyl ghrelin, which is a form lacking acyl modification
of ghrelin Ser-3, has been known to have some biological
actions such as anti-apoptotic, anti-proliferative and adipogenic
effects in mammals (Baldanzi et al., 2002; Cassoni et al., 2001,
2004; Thompson et al., 2004). Stimulatory and inhibitory
effects of des-acyl ghrelin on food intake through different
regulatory pathways have been reported (Asakawa et al., 2005;
Toshinai et al., 2006). Central and peripheral injections of des-
acyl goldfish ghrelin were found ineffective in regulating food
intake of goldfish (Matsuda et al., 2006a). However, ICV or IP
pretreatment of the des-acyl ghrelin attenuates ghrelin-induced
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122 H. Kaiya et al. / Comparative Biochemistry and Physiology, Part A 149 (2008) 109–128
food intake in a dose-dependent manner (Matsuda et al., 2006a).
The mechanisms underlying the inhibitory action of des-acyl
ghrelin is currently unknown.
6.2.2. Amphibians
There are no reports about a role of ghrelin on feeding
regulation in amphibians. However, Kaiya et al. (2006a) reported
that plasma ghrelin levels, stomach ghrelin content and stomach
ghrelin mRNA levels did not change after 5-day starvation in
bullfrog. However, ghrelin mRNA was significantly increased
after 10-day starvation, suggesting a possible involvement of
ghrelin in energy metabolism and feeding behavior in bullfrog.
6.2.3. Birds
In discordance with the orexigenic actions of ghrelin
reported in mammals and fish, ICV administration of ghrelin
decreases food intake in neonatal chicks (Furuse et al., 2001;
Saito et al., 2002a, 2005). Saito et al. (2005) revealed that the
anorectic effect of ghrelin is mediated via corticotropin-
releasing factor (CRF): ICV injections of ghrelin increased
plasma corticosterone levels in chicken and the increase in
corticosterone and anorexia was attenuated by the co-adminis-
tration of ghrelin and a CRF receptor antagonist, astressin.
Similar to what has been reported in chicken, ICV injections of
ghrelin inhibit food intake in Japanese quails (Shousha et al.,
2005). However, IP injections of high doses of ghrelin inhibited
food intake, and small doses of ghrelin were found to stimulate
food intake in Japanese quail. The reason of the different effect
by the doses injected has considered a different permeability of
ghrelin into central nervous system (Shousha et al., 2005).
IV injections of chicken ghrelin (1 nmol/100 g body weight)
decrease food intake during 1 h post-injection, as well as
respiratory quotient for 14 h post-injection in chickens
(Geelissen et al., 2006). However, Kaiya et al. (2007b) reported
no change in food intake during 2 h after IV injection of chicken
ghrelin (600 pmol/100 g body weight). The difference of
response might be due to the chicken strain used for each study:
Geelissen et al. (2006) used a broiler strain, and Kaiya et al.
(2007b) used a layer strain.
6.2.4. Reptiles
There are no reports currently available on the effects of
ghrelin in regulating food intake in reptiles. Given that ghrelin
regulates appetite in fish and birds, it will be interesting to study
the role of ghrelin in regulating energy balance reptiles.
6.3. Reproduction
Kojima et al. (1999) reported no effects of ghrelin on the
release of LH and follicle-stimulating hormone (FSH) in
dispersed rat pituitary cells, supposed no reproductive effects.
However, recent studies have indicated some involvement of
ghrelin in gonadal and reproductive function in mammals. For
example, ghrelin and/or GHS-R1a express in testicular Sertoli
and Leydig cells of rats, regulate seminiferous tubule functions
and inhibit testosterone secretion. In addition, ghrelin shows
predominant inhibitory effects at central level and on GnRH-
induced gonadotropin secretion, but direct stimulatory actions
on basal LH and FSH secretion by long-term treatment (Garcia
et al., 2007; Tena-Sempere, 2005a,b).
6.3.1. Fish
There are some reports that suggest possible involvement of
ghrelin in reproduction in non-mammalian vertebrates. Plasma
ir-ghrelin levels were found to increase after spawning in burbot
(Mustonen et al., 2002) although the functional significance of
this is currently unknown. They have suggested stimulation of
gluconeogenesis by ghrelin-released GH and increase in
appetite in post-spawning fish. A sexually dimorphic expression
of ghrelin in the stomach of the Nile tilapia has been reported
(Parhar et al., 2003), pointing towards a linkage between ghrelin
and the reproductive axis. In goldfish, ghrelin stimulates LH
release and LH-β mRNA expression in pituitary cells
(Unniappan and Peter, 2004), indicating direct actions of
ghrelin on goldfish gonadotrophs. The minimal effective dose
of ghrelin is comparable to that of gonadotropin-releasing
hormone (GnRH) and NPY (Marchant et al., 1989; Peng et al.,
1993), suggesting physiological importance of ghrelin on the
regulation of LH release and synthesis in goldfish. Further
studies, including those examining the role of steroid hormones
in the regulation of ghrelin synthesis and release, will be helpful
in unraveling the role of ghrelin in the reproductive physiology
of fish.
6.3.2. Birds
Yoshimura et al. (2005) identified ir-ghrelin and its mRNA in
the oviduct of laying Japanese quails. Ghrelin immunoreactivity
and mRNA expression were also found in the infundibulum and
magnum, but not in the isthmus, uterus and vagina. The intensity
of ghrelin immunoreactivity in the infundibulum and magnum
reduced post-ovulation, suggesting secretory activity of ghrelin
during the passage of eggs through oviduct. The physiological
relevance remains unknown, but a possible involvement of
ghrelin in embryonic development through GH has been
proposed (Decuypere and Buyse, 2005). Furthermore, Sirotkin
and colleagues have reported possible involvement of ghrelin in
the control of gonadal functions in chicken, such as proliferation,
apoptosis, and hormone secretion (Sirotkin et al., 2006). Ghrelin
and GHS-R1a mRNA expression was found in chicken ovarian
tissue and cultured chicken ovarian cells. A ghrelin analog 1–18
increased makers of proliferation (PCNA, cyclin) and decreased
makers of apoptosis (caspase-3, bax, bcl-2 and TUNEL-positive
cells) in ovarian granulosa cells. In chicken ovarian follicular
fragments, ghrelin 1–18 increased PCNA, cyclin, bax and p53,
where ghrelin stimulated the release of progesterone, estradiol,
arginin–vasotocin (AVT) and IGF-1, but not testosterone. These
secretory activities of ghrelin on ovarian cell are mediated through
TK-, MAPK-, CDK- or OKA-dependent intracellular mechan-
isms (Sirotkin and Grossmann, 2007).
6.4. Gastrointestinal motility
Ghrelin promotes gastric emptying in mammals when
injected centrally and peripherally. The motor effects of ghrelin
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H. Kaiya et al. / Comparative Biochemistry and Physiology, Part A 149 (2008) 109–128
are regulated by efferent pathways from the central nervous
system, vagal afferents and the enteric nervous system (Peeters,
2005). There are two reports about the effect of ghrelin on
gastrointestinal motility in chickens. In layer chicks, ICV
injections of a peptidyl GHS, GHRP-6 did not affect
gastrointestinal motility (Khan et al., 2006). Since GHS inhibits
food intake in neonatal chicks when injected centrally (Saito
et al., 2002a,b; Khan et al., 2006), these results were found
supportive for the anorexigenic actions. However, Kitazawa
et al. (2007) clearly demonstrated different actions of various
GHS-R agonists on contractile effect on the chicken gastro-
intestinal tract. Homologous chicken ghrelin stimulated the
contraction of the crop and proventriculus, but GHRP-6 and rat
and human ghrelins showed only weak stimulatory effects.
Kitazawa at al. (2007) also showed that chicken ghrelin caused
contractile responses in smooth muscle strips isolated from
several parts of the gastrointestinal tract such as esophagus,
crop, proventriculus, duodenum, jejunum and colon, and that
localization of functional GHS-R in the gastrointestinal wall is
region- and species-specific. In addition, chicken ghrelin and
motilin, a ghrelin-related gut peptide, both caused contraction of
chicken gastrointestinal tract with a region-dependent manner:
ghrelin mainly stimulated the contractions of upper (crop) and
lower (colon) intestines, while motilin principally stimulated
those of middle and small intestines. It will be interesting to
compare the effects of ghrelin on the motility of different parts
of the gastrointestinal tracts of other non-mammalian
vertebrates.
6.5. Regulation of water intake
Kozaka et al. (2003) demonstrated that the intracranial
injection of ghrelin inhibited water intake in the seawater-
adapted eel. This result provides further evidence for the central
actions of ghrelin in fish. Furthermore, it has been shown that
ghrelin potently inhibits water intake in neonatal chicks when
injected ICV (Tachibana et al., 2006). Recently, similar
inhibitory effects on water intake have been reported in rats
(Hashimoto et al., 2007), via actions on the area postrema (AP)
and the nucleus of the tractus solitarius (NTS). These results
from fish to mammals indicate that regulation of water intake by
ghrelin may be a conserved physiological action in vertebrates.
6.6. Immunity
In mammals, ghrelin participates in immune functions.
Hattori et al. (2001) has reported expression of the GHS-R gene
in human T cells, B cells and neutrophils. Dixit and Taub (2005)
revealed that ghrelin inhibits expression of proinflammatory
cytokine in human T cells and monocytes. These results suggest
that ghrelin acts on the immune cells through GHS-R.
In non-mammalian vertebrates, Yada et al. (2006) demon-
strated actions by ghrelin on the fish immune system using
rainbow trout. Ghrelin stimulates superoxide production and
activates phagocytosis in leukocytes derived from the head
kidney of rainbow trout. Interestingly, these effects were found
mediated by GH, secreted from leucocytes in response to
123
ghrelin. It is possible that similar regulatory systems are present
among vertebrates.
6.7. Growth effects
Ghrelin mRNA expression increases before the “rapid
growth period” in Nile tilapia, suggesting its direct role in the
growth of tilapia (Parhar et al., 2003). It has been reported that
ghrelin affects IGF-I release in Mozambique tilapia and rainbow
trout (Fox et al., 2007; Shepherd et al., 2007). In rainbow trout,
plasma levels of the 32 kDa, 42 kDa and 50 kDa IGF binding
protein (IGFBP) vary after KP102 and rat ghrelin (Shepherd et
al., 2007), suggesting a ghrelin action on the somatotropic axis.
Continuous infusion of tilapia ghrelin C-10 but not C-8 for
21 days using osmotic mini-pumps resulted in increases in liver
weight, total lipid content in the liver and muscle of
Mozambique tilapia although the longitudinal growth did not
increase and plasma IGF-1 levels were decreased (Riley et al.,
2005). The reduction of fat mobilization and stimulation of
lipogenesis are similar with that observed in rodents (Tschöp et
al., 2000). In chicken, bolus IV injections of ghrelin had no
apparent effects on lipid metabolism, as plasma glucose,
triglyceride, free fatty acids protein levels, as well as
thermogenesis were unaffected (Geelissen et al., 2006).
6.8. Behavior
In goldfish, ICV or IP injection of octanoylated 12-amino
acid goldfish ghrelin increases locomotor activity of goldfish,
although des-acyl ghrelin has no effect, suggesting specific
actions of the ghrelin–GHS-R system on locomotor behavior of
fish (Matsuda et al., 2006a,b).
In chicken, ICV injections of ghrelin induced hyperactiv-
ity and a sleep-like behavior (Tachibana et al., 2001; Saito et
Fig. 5. Multiple physiological functions of ghrelin in non-mammalian
vertebrates. Biological actions that ghrelin was found to regulate at least in
one non-mammalian vertebrate have been included. Information used to develop
this figure was obtained from published articles on ghrelin in non-mammalian
vertebrates, and in some instances based on unpublished, preliminary
observations.
 Author's personal copy
124 H. Kaiya et al. / Comparative Biochemistry and Physiology, Part A 149 (2008) 109–128
al., 2002b). These behaviors are contradictory to each other
but are observed during a different time course: hyperactivity
was induced during the first 30 min, and then sleep-like
behavior occurred during the next 30 min. The hyperactivity
is due to the activation of hypothalamic CRF system by
ghrelin in relation to inhibition of food intake (Saito et al.,
2005).
7. Perspectives
There is enough evidence available now to highlight the
importance of ghrelin as a multifaceted endocrine regulator in
non-mammalian vertebrates (Fig. 5). The high similarity in the
structure of ghrelin gene, peptide and GHS-R1a, distribution
pattern of ghrelin and GHS-R1a and the commonality of many of
ghrelin's functions among vertebrates indicate that ghrelin is an
evolutionarily conserved, essential hormone in vertebrates. While
ghrelin shares some biological functions across the vertebrates, it
is also clear that some functions are more species-specific. Based
on the results reported so far, it is reasonable to conclude that the
regulation of GH release and feeding would be two major
functions of ghrelin in vertebrates. Future studies employing
cellular, molecular and physiological tools and in vitro and in
vivo approaches could enable us to better understand the biology
of ghrelin, not only in non-mammalian vertebrates but also in all
vertebrates. Some of the properties of ghrelin, especially its role
in the regulation of food intake and GH release are currently
being explored to develop potential therapies for the treatment of
several endocrine pathophysiologies in humans. Utilizing the
potential of the effects of ghrelin on feeding and pituitary
hormone release in non-mammalian vertebrates could also be
beneficial, for example, in poultry and aquaculture to promote
growth and weight gain of cultured species.
8. Summary
In this review, we discussed many aspects of ghrelin from
fish to birds, in comparison to mammals. Ghrelin is a conserved
hormone during vertebrate evolution and have multiple
fundamental biological effects. Research on non-mammalian
ghrelin is at its infancy. Comparative endocrinological
approaches using non-mammalian vertebrates would act as
strong tools to better understand the commonality of ghrelin in
vertebrates.
Acknowledgments
SU is funded by a Discovery Grant and a Research Tools and
Instruments Grant from the Natural Sciences and Engineering
Research Council (NSERC) of Canada. REP received funding
from the NSERC of Canada. HK, MM and KK are supported by
a Grant-in-Aid for Scientific Research from the Ministry of
Education, Culture, Science, Sports and Technology (MEXT) of
Japan, the Ministry of Health, Labor and Welfare of Japan, and
the Promotion of Fundamental Studies in Health Science from
the Organization for Pharmaceutical Safety and Research
(ORSR) of Japan.
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