Multiple Forms Of Gonadotropin-releasing Hormone Research Articles

Gonadotropin releasing hormone (GnRH) and gonadotropin inhibitory hormone (GnIH) in the songbird hippocampus: Regional and sex differences in adult zebra finches

Hypothalamic gonadotropin releasing hormone (GnRH) and gonadotropin inhibitory hormone (GnIH) are vital to reproduction in all vertebrates. These neuropeptides are also present outside of the hypothalamus, but the roles of extra-hypothalamic GnRH and GnIH remain enigmatic and widely underappreciated. We used immunohistochemistry and PCR to examine whether multiple forms of GnRH (chicken GnRH-I (GnRH1), chicken GnRH-II (GnRH2) and lamprey GnRH-III (GnRH4)) and GnIH are present in the hippocampus (Hp) of adult zebra finches (Taeniopygia guttata). Using immunohistochemistry, we provide evidence that GnRH1, GnRH2 and GnRH4 are present in hippocampal cell bodies and/or fibers and that GnIH is present in hippocampal fibers only. There are regional differences in hippocampal GnRH immunoreactivity, and these vary across the different forms of GnRH. There are also sex differences in hippocampal GnRH immunoreactivity, with generally more GnRH1 and GnRH2 in the female Hp. In addition, we used PCR to examine the presence of GnRH1 mRNA and GnIH mRNA in micropunches of Hp. PCR and subsequent product sequencing demonstrated the presence of GnRH1 mRNA and the absence of GnIH mRNA in the Hp, consistent with the pattern of immunohistochemical results. To our knowledge, this is the first study in any species to systematically examine multiple forms of GnRH in the Hp or to quantify sex or regional differences in hippocampal GnRH. Moreover, this is the first demonstration of GnIH in the avian Hp. These data shed light on an important issue: the sites of action and possible functions of GnRH and GnIH outside of the HPG axis.

Molecular characterization and expression profiles of three GnRH forms in the brain and pituitary of adult chub mackerel (Scomber japonicus) maintained in captivity

Gonadotropin-releasing hormone (GnRH) is a key neuroendocrine peptide involved in the reproduction of fish and other vertebrates. However, characterizing the involvement of GnRH in fish reproduction has been complicated by the discovery of multiple GnRH forms. In the present study, we isolated full-length cDNAs encoding three GnRH forms and analyzed seasonal changes in the concentrations of mRNA in the brain and corresponding peptides in the brain and pituitary, in relation to seasonal gonadal development of chub mackerel (Scomber japonicus). Chub mackerel sbGnRH, cGnRH-II, and sGnRH cDNAs encode 98, 85, and 90 deduced amino acids, respectively. In females, brain sbGnRH mRNA and peptide concentrations were significantly higher only during the post-spawning season (August); however, pituitary peptide concentrations were higher during late vitellogenesis (April) and the post-spawning season, in comparison to immature stage (November). In males, brain sbGnRH mRNA and pituitary peptide concentrations were higher during spermiation (April). No significant differences in cGnRH-II mRNA or peptide concentrations were found in either sex. Furthermore, in females, brain sGnRH mRNA concentrations did not vary significantly; however, corresponding peptide concentrations in the brain and pituitary were higher during late vitellogenesis and the post-spawning season, respectively. In males, only brain sGnRH mRNA concentrations were higher during the post-spawning season, with no significant change in peptide concentrations. This study quantified the seasonal expression changes of three GnRH mRNAs and peptides in both sexes of chub mackerel, and the present results combined with our previous immunocytochemical report indicates that sbGnRH form plays a dominant role in seasonal gonadal development.

Introduction to special topic—estrogenic control of hypothalamic GnRH neurons

In female mammals, sexual maturation is initiated by a change in the release pattern of pituitary gonadotropins, luteinizing hormone (LH) and follicle-stimulating hormone (FSH), and a consequential increase in the secretion of ovarian sex-steroids, estradiol and progesterone. Orchestrating these endocrine changes is a diffuse population of hypothalamic neurons that produce the neuropeptide, gonadotropin-releasing hormone (GnRH). Although the development and functional integration of GnRH neurons with the rest of the central nervous system is still poorly understood, recent progress has been made on several fronts. For example, there is now evidence that some mammalian species express more than one molecular form of GnRH (GnRH-I and GnRH-II), and that the two corresponding GnRH neuronal sub-populations may play different roles in the regulation of reproductive function and behavior. Moreover, through the use of transgenic animal models, neuronal fiber tracing, gene expression profiling, and electrophysiologcal recordings, new insights have been gained into the mechanisms that regulate GnRH release. This volume brings together 10 articles that reflect current thinking about the role of GnRH in mediating estrogenic feedback to the neuroendocrine reproductive axis. The main focus is on the negative and positive actions that estrogens exert within the hypothalamus of mammals, especially around the time of the preovulatory LH surge. However, the articles also present findings from other vertebrate classes, and so provide an intersting comparative perspective. Taken together, the knowledge presented in this volume represents a foundation upon which to develop more effective therapies for pubertal disorders, infertility, and menopause. The first article (Sower and Baron, 2011) demonstrates the expression of estrogen receptors within the lamprey hypothalamus, suggesting potential feedback interactions between esatradiol and GnRH neurons. Phylogenetically, lampreys are positioned as basal vertebrates, and like many other non-mammalian vertebrates they express multiple forms of GnRH. Consequently, studies in lampreys provide us with important insights into the molecular evolution of estrogenic-GnRH interactions. A cluster of three articles then focus on hypothalamic neuropeptides that exert a major influences on the secretion of GnRH, and which are thought to mediate the feedback effects of sex-steroids on the reproductive axis. For example, studies in rodents have clearly shown the importance of kisspeptin to both pulsatile and surge modes of GnRH release (Navarro, 2012), while human studies have observed sexual dimorphism in kisspeptin neurons during aging (Hrabovszky et al., 2011). Again, an interesting evolutionary perspective is provided by comparing the steroid sensitivity of kisspeptin neurons in various non-mammalian vertebrates (Kanda and Oka, 2012). Although the exact mechanism by which estrogens interact with the reproductive neuroendocrine axis is unclear, our general understanding of this issue is neatly summarized by a series of three reviews (Micevych and Sinchak, 2011; Kenealy and Terasawa, 2012; Radovick et al., 2012). Of particular interest is the finding that sex-steroids exert their feedback not only through classic estrogen receptors located in the nucleus, but also more rapidly through receptors located in the cell membrane. Various physiological functions show adaptations for life in an environment that undergoes marked changes across the 24-h day. For example, in many mammals the timing of the preovulatory LH surge occurs at a very specific time of day. Two articles discuss the interactions between the circadian timing system and estrogen-sensitive neural circuits that regulate GnRH secretion and the preovulatory surge (Sun et al., 2012; Williams and Kriegsfeld, 2012). Finally, a novel hypothesis is proposed to explain the conundrum of how estrogens appear to exert both negative and positive feedback actions on the reproductive axis (Urbanski, 2012). Data is presented from non-human primate studies to suggest that negative and positive estrogen feedback are two completely separate mechanisms, which are mediated, respectively, by two distinct GnRH neuronal populations. This hypothesis challenges the common assumption that has dominated the field for the past four decades, namely that individual GnRH neurons are relatively homogenous and can mediate both negative and positive estrogen feedback.

Immunological Characterization and Distribution of Three GnRH Forms in the Brain and Pituitary Gland of Chub Mackerel (Scomber japonicus)

The presence of three gonadotropin-releasing hormone (GnRH) forms in the brain of the chub mackerel, Scomber japonicus, namely, salmon GnRH (sGnRH), chicken GnRH-II (cGnRH-II), and seabream GnRH (sbGnRH), was confirmed by combined high performance liquid chromatography (HPLC) and time-resolved fluoroimmunoassay (TR-FIA). Immunocytochemical localization of the three GnRH forms in the brain was Investigated by using specific antisera, to elucidate possible roles of each GnRH form in reproduction in this species, and double immunolabeling was used to localize GnRH-ir (immunoreactive) fibers Innervating the pituitary. sGnRH-ir neurons were localized in the ventral olfactory bulb and terminal nerve ganglion region. Further, sGnRH-ir fibers were found in different regions of the brain, with prominent fibers running in parallel in the preoptic area (POA) without entering the pituitary. cGnRH-II-ir cell bodies were observed only in the midbrain tegmentum region, with a wide distribution of fibers, which were dense in the midbrain tegmentum and spinal cord. SbGnRH-ir cell bodies were localized in the nucleus preopticus of the POA, with fibers in the olfactory bulb, POA, and hypothalamus. Among the three GnRH forms, only SbGnRH-ir fibers innervated the pituitary gland from the preoptic-hypothalamic region, targeting follicle stimulating hormone (FSH) and luteinizing hormone (LH)-producing cells in the proximal pars distalis, as demonstrated by double immunocytochemistry. The localization of the GnRH-ir system was similar in male and female fish. These results demonstrate that multiple GnRH forms exist in the brain of the chub mackerel and suggest that they serve different functions, with SbGnRH having a significant role in reproduction in stimulating FSH- and LH-producing cells, and sGnRH and cGnRH-II serving as neurotransmitters or neuromodulators.

Opportunism, photoperiodism, and puberty: Different mechanisms or variations on a theme?

There are many parallels between the neural regulation of seasonal breeding in birds and puberty in primates, but most studies of the regulation of puberty in vertebrates involve the use of rodents. The findings from rodent studies are often perceived as being typical of mammals and therefore pertinent to human reproductive biology and in many cases, rodent models have a great deal to offer in terms of an understanding of the regulation of puberty and reproductive biology. However, knowledge available from comparative work perhaps highlights mechanistic similarities that may not exist between rodent and primate systems. In this short review, we highlight some of the advantages of studying avian reproductive biology in this regard. We discuss disparities between rodent puberty and primate puberty, and similarities between primates and birds. Thus, understanding the mechanisms regulating avian puberty and seasonal breeding might in some cases provide greater insight into the mechanistic control of puberty in nonrodent mammals. We also describe in detail the neuroendocrine regulation of reproduction in birds and aim to provoke discussion of the possible roles of thyroid hormones and multiple forms of gonadotropin-releasing hormone in avian and mammalian reproduction.

Dimorphic expression of GnRH, gonadotropin and tryptophan hydroxylase provides new markers to follow brain sex development in teleosts

resence of multiple forms of gonadotropin-releasing hormone (GnRH) is comon in teleosts. Among these, species-specific form of GnRH plays an important ole in regulating gonadotropin (GTH) in turn reproduction. Interestingly, serotonin 5-HT) plays an important role to regulate GnRH and GTH release. Nevertheless, ole of GnRH, GTH and 5-HT has not been implicated in sex differentiation of eleosts. Studies from our laboratory addressed this issue to some extent. Genetic Y males have differential seabream GnRH immunoreactivity (-ir) while in females teady -ir was evident during early development. It is true with reference to GTH. ur studies on tryptophan hydroxylase, rate limiting enzyme for 5-HT biosynthesis lso showed male-specific expression in teleosts. Based on these, we presume that uitable markers can be developed to identify male brain sex differentiation.

Molecular co‐evolution of Gonadotropin‐releasing hormones and their receptors

Abstract Gonadotropin‐releasing hormone (GnRH), synthesized in the hypothalamus, plays a pivotal role in the regulation of vertebrate reproduction. Since molecular isoforms of GnRH and their receptors (GnRHR) have been isolated in a broad range of vertebrate species, GnRH and GnRHR provide an excellent model for understanding the molecular co‐evolution of a peptide ligand‐receptor pair. Vertebrate species possess multiple forms of GnRH, which have been created through evolutionary mechanisms such as gene/chromosome duplication, gene deletion and modification. Similar to GnRHs, GnRH receptors (GnRHR) have also been diversified evolutionarily. Comparative ligand‐receptor interaction studies for non‐mammalian and mammalian GnRHRs combined with mutational mapping studies of GnRHRs have aided the identification of domains or motifs responsible for ligand binding and receptor activation. Here we discuss the molecular basis of GnRH‐GnRHR co‐evolution, particularly the structure‐function relationship regarding li...

Chromosomal Organization, Evolutionary Relationship, and Expression of Zebrafish GnRH Family Members

Multiple forms of gonadotropin-releasing hormone (GnRH) are found in different vertebrates. In this study, we have cloned cDNA encoding the full-length gnrh3 and gnrh2 from zebrafish brain and characterized their structure and expression patterns. We performed phylogenetic analysis and compared conserved syntenies in the region surrounding the GnRH genes from human, chicken, pufferfish, and zebrafish genores. The gnrh3 and gnrh2 genes were mapped to LG17 and LG21, respectively. The zebrafish genome appears to lack an ortholog to human GNRH1, and the human genome appears to lack an ortholog of gnrh3. Expression of gnrh3 began in the olfactory pit at 24-26 h postfertilization and expanded to the olfactory bulb during early larval stage. Expression of gnrh2 is always in the midbrain. In addition, GnRH is also expressed in boundary cells surrounding seminiferous cysts of the testis. Thus, this detailed phylogenetic, chromosomal comparison, and expression study defines the identity and the evolutionary relationship of two zebrafish gnrh genes. We propose a model describing the evolution of gnrh genes involving ancestral duplication of the genes followed by selective loss of one gene in some teleosts.

Gonadotropin-Releasing Hormone (GnRH) Variants in a Lizard Brain: Is Mammalian GnRH Being Expressed?

In reptiles as in other vertebrates, multiple forms of gonadotropin-releasing hormone (GnRH) within a single brain have been identified. In this group the following GnRH molecular variants have been characterized either by direct or indirect methods: chicken GnRH I (cGnRH-I), chicken GnRH II (cGnRH-II), salmon GnRH (sGnRH) and several unidentified GnRH-like forms. In the present study GnRH variants were investigated in brain extracts of the lizard Tupinambis teguixin (= T. merinae) by combining high-performance liquid chromatography (RP-HPLC) followed by radioimmunoassays (RIA). Two peaks showing GnRH immunoreactivity with the elution position of synthetic mammalian GnRH (mGnRH) and cGnRH-II were detected. Both peaks were further analyzed with different radioimmunoassay systems specific for mGnRH, cGnRH-I, and cGnRH-II. Pooled fractions corresponding to the first eluting peak showed no crossreactivity when analyzed with a cGnRH-I specific assay and logit-log displacement curves were not significantly different from those of synthetic mGnRH with homologous RIA systems. The second peak showed immunological characteristics of cGnRH-II when analyzed with a specific antiserum. The first ir-GnRH peak was selected for further RP-HPLC purification showing similar chromatographic behavior as mGnRH synthetic standard. We demonstrated the absence of cGnRH-I in this lizard using well-characterized antisera.

Distribution of immunoreactive multiple forms of gonadotropin-releasing hormone in the brain of the antarctic fish, Notothenia coriiceps

Previous studies have shown that different gonadotropin-releasing hormone (GnRH) molecular forms are present in different groups of bony fish. In the present study, we have investigated the possible influence of the antarctic environmental contributions upon the distribution and biochemical patterns of GnRH- related molecules. The immunocytochemical distribution of GnRH-like peptides has been studied in the brain of the antarctic fish, Notothenia coriiceps, using antisera raised against three variants of GnRH: mammalian (m-GnRH), chicken (cII-GnRH) and salmon (s-GnRH). cII-GnRH immunoreactivity appears confined to cell bodies located in the lateral hypothalamus, the ventral thalamus and the midbrain rostral tegmentum; immunoreactive nerve fibers densely innervated the hypothalamic periventricular region. By contrast, m-GnRH-like immunoreactive neurons are present exclusively in the torus semicircularis of the mesencephalon and in the outer plexiphorm layers of the optic tectum. These findings suggest that cII-GnRH-like peptides appear to function as hypophysiotropic factors, as demonstrated in other species of bony fish, whereas m-GnRH-like peptides could be involved in modulatory pathways of vestibular and visual functions of N. coriiceps. Incubation with s-GnRH antiserum failed to prove the occurrence of immunoreactive elements; consequently, at least two molecular forms related to cII-GnRH and m-GnRH seem to act as hypophysiotropic and neuromodulatory factors in the brain of Notothenia coriiceps. Moreover, m-GnRH immunoreactivity in ependymal tanycytes suggests the involvement of such specialized glial cells in neuroendocrine function by linking the cerebrospinal fluid and the median eminence, as demonstrated in mammals.

Immunoreactive Gonadotropin-Releasing Hormone (GnRH) Is Detected Only in the Form of Chicken GnRH-II within the Brain of the Green Anole,Anolis carolinensis

The presence of multiple forms of gonadotropin-releasing hormone (GnRH) within a single brain is common among vertebrate species. In previous studies of reptiles, two forms of GnRH were isolated from the brain of alligators and the primary structure was determined to be that of chicken (c)GnRH-I and cGnRH-II. GnRH has also been detected by indirect methods in other reptiles including turtles, lizards, and snakes. We used a combination of high-performance liquid chromatography (HPLC) and radioimmunoassay to determine the number and molecular form(s) of GnRH in the brain of a lizard,Anolis carolinensis,that was reported to lack GnRH cells in the forebrain. Immunoreactivity was detected in the same HPLC elution position in which synthetic cGnRH-II elutes, but not in any other position. Detection was based on five antisera that among them detect the 12 known forms of GnRH; these antisera include ones that are specific to cGnRH-I and cGnRH-II. We conclude that the lizardA. carolinensiscontains cGnRH-II, but not cGnRH-I or another known form of GnRH. These data, coupled with our earlier immunocytochemical study, suggest that the lizard studied here lacks cGnRH-I, the form that is found in the terminal nerve, olfactory bulb, and forebrain in nonsquamate reptiles and in birds. Our hypothesis is that the presence of both cGnRH-I and cGnRH-II in the brain is ancestral in the reptilian lineage and retained in the orders that include turtles (Chelonia) or alligators (Crocodilia). However, the pattern in the order Squamata varies: inA. carolinensis,only cGnRH-II is present in the brain and cGnRH-I is absent, whereas in the snakeThamnophilis sirtalis,cGnRH-I is retained and cGnRH-II is absent in the brain, as recently reported. This raises the question of how reproduction is controlled in reptiles that lack one form of GnRH.

Characterization of the Pacific salmon gonadotropin-releasing hormone gene, copy number and transcription start site

Multiple forms of gonadotropin-releasing hormone (GnRH) have been shown to exist in all vertebrates examined except recently-evolved placental mammals. To study the origin and regulation of the GnRH genes in a Pacific salmon ( Oncorhynchus nerka), we isolated and sequenced the salmon form of GnRH. The Southern blot shows a single band that strongly hybridizes to a probe for the gene reported here and weaker bands that may represent genes for related forms of GnRH. There is strong conservation of sequence in the hormone coding region and of the gene organization between fish and mammals. However, the GnRH-associated peptide (GAP) shows very little sequence identity with the mammalian GAPs, questioning its physiological role. We also show for the first time the transcriptional start site for a GnRH gene in a non-mammalian species. Interestingly, a large segment of 1152 nucleotides in the promoter region of the Pacific salmon GnRH gene is missing compared with the Atlantic salmon ( Salmo salar) gene. These gene rearrangements suggest that these two salmonid species, which have been geographically separated for 8–15 million years, have evolved promoters with different mechanisms for control and transcription of GnRH.

Three gonadotropin-releasing hormone genes in one organism suggest novel roles for an ancient peptide.

Gonadotropin-releasing hormone (GnRH) is known and named for its essential role in vertebrate reproduction. Release of this decapeptide from neurons in the hypothalamus controls pituitary gonadotropin levels which, in turn, regulate gonadal state. The importance of GnRH is underscored by its widespread expression and conservation across vertebrate taxa: five amino acids are invariant in all nine known forms, whereas two others show only conservative changes. In most eutherian mammals, only one form, expressed in the hypothalamus, is thought to exist, although in a recent report, antibody staining in developing primates suggests an additional form. In contrast, multiple GnRH forms and expression loci have been reported in many non-mammalian vertebrates. However, evidence based on immunological discrimination does not always agree with analysis of gene expression, since GnRH forms encoded by different genes may not be reliably distinguished by antibodies. Here we report the expression of three distinct GnRH genes in a teleost fish brain, including the sequence encoding a novel GnRH preprohormone. Using in situ hybridization, we show that this form is found only in neurons that project to the pituitary and exhibit changes in soma size depending on social and reproductive state. The other two GnRH genes are expressed in other, distinct cell populations. All three genes share the motif of encoding a polypeptide consisting of GnRH and a GnRH-associated peptide. Whereas the GnRH moiety is highly conserved, the GnRH-associated peptides are not, reflecting differential selective pressure on different parts of the gene. GnRH forms expressed in nonhypothalamic regions may serve to coordinate reproductive activities of the animal.

Distribution of chicken‐II gonadotropin‐releasing hormone in mammalian brain

Brains of nonmammalian vertebrates typically contain multiple forms of gonadotropin-releasing hormone (GnRH). Until recently, only the mammalian form of GnRH (mGnRH) had been isolated in placental mammals. Biochemical and histological data show that both mGnRH and chicken-II GnRH (cGnRH-II) are present in a primitive placental mammal, the musk shrew (Suncus murinus). Similar to the case in nonmammalian species, in the musk shrew, neurons that express cGnRH-II are located in a discrete cluster in the midbrain. We have used a combination of radioimmunoassay and immunocytochemistry, analyzed at the light level and with electron microscopy, to describe the distribution of cGnRH-II cell bodies and fibers in the musk shrew brain. All cGnRH-II-immunoreactive (ir) neurons reside in the midbrain, and this area contains the greatest concentration of cGnRH-II peptide in the brain. At the light and electron micrographic levels, we have identified synaptic terminals containing dense core vesicles that are immunoreactive for cGnRH-II in the medial habenula. Radioimmunoassay reveals that this region contains the second greatest concentration of cGnRH-II in the brain. Widely scattered cGnRH-II-ir fibers are present throughout the forebrain, particularly in the medial septum, hypothalamus, and midbrain central gray. Scant cGnRH-II fibers are present in the median eminence, arcuate nucleus, and infundibular stem, and only low concentrations of the peptide are detected in these areas. Finally, intravenous administration of mGnRH is ten times more effective than cGnRH-II in promoting ovulation.(ABSTRACT TRUNCATED AT 250 WORDS)

Identity of gonadotropin-releasing hormone in passerine birds: Comparison of GnRH in song sparrow ( Melospiza melodia) and starling ( Sturnus vulgaris) with five vertebrate GnRHs

Gonadotropin-releasing hormone (GnRH) was detected in the brains of passerine birds, a recently evolved and diverse avian group. The molecular forms of GnRH in two species of birds under breeding conditions were deduced using methods of HPLC and immunology. The brain extracts of song sparrows ( Melospiza melodia) contained a form of GnRH identified as chicken I GnRH-like peptide by its HPLC elution pattern and cross-reactivity with four antisera. In contrast, starling ( Sturnus vulgaris) brain extracts showed molecular heterogeneity of GnRH forms; equal amounts of chicken I and chicken II GnRH-like peptides were present. Neither bird contained GnRH that could be identified as mammalian, salmon, or lamprey GnRH. Chicken II GnRH-like peptide may not have evolved after the separation of the song sparrow and starling as both peptides are found in chicken, a more primitive bird. The possibility remains that different stages of the life cycle are associated with the expression of these GnRH-like peptides or their ratio. Only determination of the primary structure will establish whether our chromatographic and immunological evidence is correct that chicken I and II GnRH are present in passerine birds and have been conserved in representatives throughout the reptiles and birds. Starlings can be added now to the growing list of submammalian species that express multiple forms of GnRH in their brains.

Evolution of a Neuropeptide Family: Gonadotropin-Releasing Hormone

SYNOPSIS. Gonadotropin-releasing hormone (GnRH), a small peptide in the brain, is essential for reproduction. It is now clear that GnRH is part of a family of closely related molecules. The primary structure has been identified for 4 GnRH molecules: mammalian, chicken I, chicken II and salmon. During evolution the molecule has been conserved in length, terminal amino acid structure, 70–90% of amino acid sequence and the His2Trp3 residues,residues, which are important in the release of gonadotropin. Alterations have occurred in positions 5, 7 and 8, regions thought to be involved in receptor binding. The receptors for GnRH have apparently evolved also in that the mammalian and avian receptors vary considerably in their ability to bind different GnRH molecules. Other GnRH family members have been distinguished indirectly by chromatographic or immunological means; 3 different GnRH-like molecules are present, respectively, in lamprey, sturgeon and salmon (a second form). Several GnRH-like molecules including those in chondrichthyes have not yet been distinguished from the proposed salmon II molecule. The lamprey GnRH-like molecule may be a nodal point inthe analysis of the ancestral molecule; hagfish do not contain a detectable GnRH molecule. The elucidation of the GnRH precursor molecule in human placenta showed the presence of a 53-amino-acid gene-related peptide of unknown function, but did not reveal the basis for expression of multiple GnRH forms in many nonmammalian species. GnRH has a variety of novel functions in addition to release of gonadotropin from the pituitary. During evolution certain functions such as those in the retina and sympathetic ganglia have apparently disappeared in amniotes, but GnRH placental functions have appeared in mammals.

Multiple forms of gonadotropin-releasing hormone in amphibian brains

Several forms of gonadotropin-releasing hormone (GnRH)-like molecules were found in brains of both anurans (frogs) and urodeles (salamanders). The presence of the mammalian-like GnRH molecule was confirmed by HPLC and cross-reactivity studies. Small amounts of salmonid-like GnRH molecules in the brains of frogs ( Rana pipiens, Hyla regilla) and salamanders ( Taricha granulosa, Ambystoma gracile) were detected by comparing the HPLC chromatographic pattern and immunological reactivity of the brain extracts with native trout and synthetic salmon GnRH. This nonmammalian form of GnRH in the amphibian brain is similar and perhaps identical, at least by indirect evidence, to a form of GnRH reported earlier to be in sympathetic ganglion, retina, chromaffin tissue, and tadpole brain. If two of the amphibian GnRH molecules prove to be mammalian and salmon GnRH, then it is likely that two separate genes in amphibians code for the distinct primary structures of the molecules. The most parsimonious interpretation of the presence of both mammalian- and salmon-like GnRH in anurans and urodeles is that a common phylogenetic ancestor also possessed the two forms of GnRH. Thus the mammalian form of GnRH may well have been present in labyrinthodont amphibians. Independent of evolutionary origin, the functions of the different GnRH molecules in amphibians are unknown.