Forms Of Gonadotropin-releasing Hormone Research Articles

Chimaeric gonadotropin-releasing hormone (GnRH) peptides with improved affinity for the catfish (Clarias gariepinus) GnRH receptor.

The gonadotropin-releasing hormone (GnRH) receptor in catfish differs from its mammalian counterparts in showing a very low affinity for the hypothalamic GnRH form [i.e. catfish GnRH (cfGnRH)] and a very high affinity for the highly conserved mesencephalic GnRH, chicken GnRH-II (cGnRH-II). In the present study we investigated the molecular interactions between ligand and receptor involved in determining the ligand selectivity of the catfish GnRH receptor. Studies on the binding characteristics of the catfish GnRH receptor for cfGnRH and cGnRH-II as well as for mammalian GnRH (mGnRH) and synthetic chimaeric GnRHs, differing at positions 5, 7 and 8, revealed that the low affinity of the catfish receptor for cfGnRH can be improved by replacing Leu(7) by a tryptophan residue and/or Asn(8) by either a tyrosine or an arginine residue. Testing cfGnRH and cGnRH-II as well as mGnRH and the chimaeric GnRHs on Asp(304)-->Ala, Asp(304)-->Glu and Asp(304)-->Asn mutant catfish GnRH receptors revealed that Asp(304) of the catfish receptor mediates the recognition of Arg(8) in mGnRH, as well as in the chimaeric peptides [Arg(8)]cfGnRH and [Arg(8)]cGnRH-II, but seems to be less important for the recognition of Tyr(8) in cGnRH-II. On the basis of these results, a three-dimensional model for the binding of [Arg(8)]cGnRH-II to the catfish GnRH receptor is proposed.

Gonadotropin-releasing hormone (GnRH): from fish to mammalian brains.

This work deals with a family of neuropeptides, gonadotropin-releasing hormone (GnRH), that play a key role in the development and maintenance of reproductive function in vertebrates. 2. Until now, a total of 16 GnRH structural variants have been isolated and characterized from vertebrate and protochordate nervous tissue. All vertebrate species already investigated have at least two GnRH forms coexisting in the central nervous system. However, it is now well accepted that three forms of GnRH in early and late evolved bony fishes are present. 3. In these cases, cGnRH-II is expressed by midbrain neurons, a species-specific GnRH is present mainly in the preoptic area and the hypothalamus, and sGnRH is localized in the terminal nerve ganglion (TNG). In this context it is possible to think that three GnRH forms and three GnRH receptor (GnRH-R) subtypes are expressed in the central nervous system of a given species. 4. Then it is possible to propose three different GnRH lineages expressed by distinct brain areas in vertebrates: (1) the conserved cGnRH-II or mesencephalic lineage; or (2) the hypothalamic or "releasing" lineage whose primary structure has diverged by point mutations (mGnRH and its orthologous forms: hrGnRH, wfGnRH, cfGnRH, sbGnRH, and pjGnRH); and (3) the telencephalic sGnRH form. Also different GnRH nomenclatures are discussed.

GnRH in the invertebrates: an overview

Publisher Summary This chapter provides an overview of GnRH in the invertebrates. The gonadotropin-releasing hormone (GnRH), which was previously called luteinizing hormone-releasing hormone (LHRH), represents a pivotal peptide in animal reproduction. GnRH is a decapeptide originally isolated from the porcine and ovine hypothalamus and is characterized for its ability to enhance the release of pituitary gonadotropins, follicle-stimulating hormone (FSH), and luteinizing hormone (LH). All known forms of GnRH peptides have in common a pyroglutamyl residue at the amino terminus and an amidated glycine at the carboxy terminus. Except for the octoGnRH, all other known forms are decapeptides and differ by one or more amino acids; amino acids 4 and 9 are conserved in all forms known to date. Studies involving an evolutionary approach to GnRH have adopted one particular strategy; attention is on understudied invertebrate taxonomic groups. Invertebrate species that vary in their phylogenetic relationships are likely to reveal differences, as well as similarities, in GnRH form and function. Studies on invertebrate species are beginning to provide further insights on the structural and functional similarities and differences. The perspective that evolution has acted to shape the GnRH molecule effectively leads to investigations in a comparative approach.

Differential in vitro secretion of gonadotropin-releasing hormone (GnRH) and [hydroxyproline]GnRH from the rat hypothalamus during postnatal development.

The differential secretion of gonadotropin-releasing hormone (GnRH) and [hydroxyproline9]GnRH (HypGnRH) has been recently reported from the adult rat hypothalamus. We report here in vitro cosecretion of HypGnRH and GnRH by the hypothalamus of 2-45 day-old-rats and provide evidence that they are differentially regulated throughout development. The secretion of both forms of GnRH was increased in a dependent manner during depolarization by high K+ solutions, and was stimulated by forskolin and 12-O-tetradecanoylphorbol-13-acetate (TPA), activators of adenylate cyclase and protein kinase C pathways, respectively. The proportion of HypGnRH in the release of GnRH-like peptides remained stable and high (33-40%) under basal and K+-induced conditions until days 13 and 21, respectively. By contrast, the proportion of HypGnRH in the total GnRH-like content of the developing hypothalamus continuously decreased (from 37% to 14%). Similarly, the proportion of HypGnRH: total GnRH-like material released remained stable in TPA- (30%) and forskolin- (50%) induced secretion until postnatal day 8. Evaluation of release over tissue store ratios revealed a 1.3-to 2.8-fold higher release of HypGnRH compared to GnRH according to the different secretions and postnatal periods examined. The preferential recruitment of HypGnRH was maintained under basal and K+ conditions during postnatal development, but it disappeared under TPA stimulation from day 13 onwards. After forskolin stimulation, the preferential mobilization of HypGnRH was markedly reduced from day 2 to day 13 but recovered its high perinatal level during puberty. Taken together, our results support the hypothesis that HypGnRH may play a specific role in development. In addition, a specific function of this peptide taking place during puberty through the activation of the adenylate cyclase pathway is suggested.

Induction of Gamete Release by Gonadotropin-Releasing Hormone in a Protochordate, Ciona intestinalis

Gonadotropin-releasing hormone (GnRH) of vertebrates is now believed to have multiple functions in addition to its role as a hypophysiotropic hormone, as originally defined. Recently, it has been shown that GnRH occurs also in the ascidians, which are considered ancestral chordates. Here the author shows that GnRH induces spawning of gametes from mature individuals of Ciona intestinalis. Ciona accumulates mature gametes in the gonoducts and maintains them until spawning is triggered by a photoperiodic cue(s). Injection of synthetic tunicate GnRH-I or -II into various sites of mature individuals effectively induced gamete release (spawning), although the former was more potent. Gamete release often occurred on a larger scale than in spontaneous spawning. However, moderate gamete release, similar to spontaneous spawning, was often triggered by exogenous tunicate GnRH. GnRH in vivo apparently is released from the GnRH-containing neurons that are distributed from the region of the cerebral ganglion to the proximal part of the ovary along the dorsal strand within the blood sinus; this indicates that both forms of tunicate GnRH may be the actual inducers of spawning. It is suggested that, in the ancestral chordate, GnRH neurons release GnRH prior to the spawning and the released GnRH acts directly on the epithelium of gonoducts or functions as a neuromodulator of other neurons innervating the gonoducts to induce spawning.

Gonadotropin-Releasing Hormones (GnRHs) in a Primitive Teleost, the Arowana: Phylogenetic Evidence That Three Paralogous Lineages of GnRH Occurred Prior to the Emergence of Teleosts

Multiple molecular forms of gonadotropin-releasing hormone (GnRH) are present in a single vertebrate species. To extend the knowledge on GnRH evolution and the number of GnRH forms in one organism, GnRH cDNAs have been isolated and characterized from one of the most primitive teleosts, the arowana Scleropages jardini. This species had two molecular forms of GnRH: salmon-type GnRH (sGnRH) and chicken-II-type GnRH (cGnRH-II). Sequence comparison between the prepro-GnRHs of the arowana and those of other teleosts indicated that sGnRH represented a paralogue separate from any other forms of GnRH. Consistently, subsequent phylogenetic analysis showed that known forms of GnRH in teleosts fell into three paralogous lineages: sGnRH alone on one lineage, cGnRH-II on another, and many other forms on the other. These results suggest that an ancestral GnRH gene duplicated twice prior to the emergence of teleosts and, therefore, that teleosts, and probably also tetrapods, would possess three paralogous forms of GnRH in individual brains.

Expression and antiproliferative effect of a second form of gonadotropin-releasing hormone in normal and neoplastic ovarian surface epithelial cells.

Epithelial ovarian carcinomas are the most common cause of death from gynecological malignancies and appear to arise from ovarian surface epithelium (OSE), but the exact mechanism of ovarian tumorigenesis has not been elucidated. Recent cloning of a second form of gonadotropin-releasing hormone (GnRH-II) has been reported in various human tissues including the ovary. However, the expression and role of GnRH-II in human OSE and ovarian carcinomas is not known. In the present study, we demonstrated that in addition to the GnRH receptor (GnRH-R), GnRH-II mRNA is expressed in normal OSE, immortalized OSE (IOSE) cells, primary cultures of ovarian tumors and ovarian cancer cell lines. Treatments with increasing doses (10(-9)-10(-7) M) of GnRH-I and -II resulted in a growth-inhibition in both non-tumorigenic IOSE-29 and tumorigenic IOSE-29EC cells. These results indicate for the first time the expression and potential anti-proliferative effect of GnRH-II, suggesting that GnRH-II, similar to GnRH-I, may have a growth-regulatory effect in normal and neoplastic OSE cells.

Expression and Antiproliferative Effect of a Second Form of Gonadotropin-Releasing Hormone in Normal and Neoplastic Ovarian Surface Epithelial Cells

Epithelial ovarian carcinomas are the most common cause of death from gynecological malignancies and appear to arise from ovarian surface epithelium (OSE), but the exact mechanism of ovarian tumorigenesis has not been elucidated. Recent cloning of a second form of gonadotropin-releasing hormone (GnRH-II) has been reported in various human tissues including the ovary. However, the expression and role of GnRH-II in human OSE and ovarian carcinomas is not known. In the present study, we demonstrated that in addition to the GnRH receptor (GnRH-R), GnRH-II mRNA is expressed in normal OSE, immortalized OSE (IOSE) cells, primary cultures of ovarian tumors and ovarian cancer cell lines. Treatments with increasing doses (10−9–10−7 M) of GnRH-I and -II resulted in a growth-inhibition in both non-tumorigenic IOSE-29 and tumorigenic IOSE-29EC cells. These results indicate for the first time the expression and potential anti-proliferative effect of GnRH-II, suggesting that GnRH-II, similar to GnRH-I, may have a growth-regulatory effect in normal and neoplastic OSE cells.

A second form of gonadotropin-releasing hormone (GnRH II) activates the MMPs/TIMP-1 cascade in human decidual cells in vitro.

Objective: Implantation in the human is dependent upon the invasion of embryonic trophoblasts into the decidua of the maternal endometrium. The regulated expression of matrix metalloproteinases (MMPs) and their tissue specific inhibitors (TIMPS) have been shown to play a key role in human implantation by modulating the invasive capacity of the trophoblasts and/or mediating the cellular remodelling processes that occur in the endometrium in preparation for the implanting embryo. To date, the factors capable of regulating the expression of MMPs and TIMPs remain poorly characterised. In the present study, we have examined the ability of a novel form of Gonadotropin-Releasing Hormone (GnRH II), that is expressed in the human placenta, to regulate MMP/TIMP mRNA levels in primary cultures of human decidual cells.

The effect of gonadotropin-releasing hormone on growth hormone and gonadotropin subunit gene expression in the pituitary of goldfish, Carassius auratus

The goldfish brain contains at least two forms of gonadotropin-releasing hormone (GnRH): sGnRH and cGnRH-II. In goldfish sGnRH and cGnRH-II are present both in the brain and pituitary, and exert direct effects via specific GnRH receptors stimulating growth hormone (GH) and gonadotropin hormone (GtH) synthesis and secretion. In this study, we investigated the effects of sGnRH and cGnRH-II on GtH subunit (α, FSH-β and LH-β) and GH mRNA levels in the goldfish pituitary in vivo and in vitro. Injection of goldfish with sGnRH or cGnRH-II (4 μg/fish) stimulated GtH-α, FSH-β and LH-β mRNA levels after 24 h. For in vitro studies, goldfish pituitary fragments were treated continuously for 12 h with 10−7 M sGnRH or cGnRH-II. Both sGnRH and cGnRH-II stimulated GtH-α, FSH-β, LH-β and GH mRNA levels, however, cGnRH-II appeared to have a more pronounced effect. Similar experiments were carried out using cultured dispersed goldfish pituitary cells. In this study, treatments for 12 h with 10−7 M sGnRH or cGnRH-II also stimulated GtH and GH gene expression. The present results provide a basis for the investigation of the signal transduction pathways that mediate GnRH-induced changes in GtH subunit and GH mRNA levels in the goldfish pituitary.

Seasonal fluctuations in pituitary levels of the three forms of gonadotropin-releasing hormone in striped bass, Morone saxatilis (Teleostei), during juvenile and pubertal development.

In most vertebrates, the development of a mature gonadotropin-releasing hormone (GnRH) secretory system is pivotal for the onset of puberty. The role of the three native GnRH forms, seabream (sb) GnRH, chicken (c) GnRH-II and salmon GnRH, in striped bass puberty remains elusive. This study examined the changes in pituitary GnRH levels throughout juvenile and pubertal development, a period encompassing 3 to 4 years. The levels of the two most abundant forms in the pituitary, sbGnRH and cGnRH-II (10:1), increased during the Fall and peaked prior to (cGnRH-II) or during (sbGnRH) the natural breeding season in March to May. In most cases, sbGnRH and cGnRH-II levels of maturing fish correlated to changes in oocyte diameter, gonadosomatic index and LH pituitary content. Interestingly, pituitaries of immature and maturing 2- and 3-year-old males and females contained similar amounts of all three GnRH forms. Additionally, pituitary sbGnRH and cGnRH-II levels in juvenile fish were relatively high and GnRH profiles showed a clear seasonality, similar to those of older, mature fish. These findings suggest a role for both sbGnRH and cGnRH-II in the regulation of gonadal development and indicate that, unlike some mammalian species, the timing of puberty in striped bass is not limited by a low activity of the GnRH system.

Recombinant perciform GnRH-R activates different signaling pathways in fish and mammalian heterologous cell lines

Perciforms have three forms of gonadotropin-releasing hormone (GnRH) in their brain. All three GnRHs are potent secretogogues for luteinizing hormone (LH) from the pituitary. The pivotal role of GnRH-R–GnRH interactions in reproductive homeostasis is well established; however, there is a paucity of information on how a GnRH-R responds to the three endogenous GnRH forms in a perciform species. In this study, a recombinant pituitary GnRH-R from striped bass (stb) was expressed in a mammalian cell line (COS-7) and a fish cell line (CHSE–214). Activation of the signaling pathways was monitored by reporter gene (luciferase) based assays, which were specific for cAMP-PKA or Ca 2+/calmodulin kinase (activated via c-fos promoter) signaling pathways. The stbGnRH-R expressed in two different cell lines triggered different downstream signaling in response to the treatments with chicken (c) GnRH II. Interestingly, when endogenous GnRHs were used in combinations, the luciferase activity was significantly attenuated in transfected CHSE-214 cells.

A Developmental Increase in the Expression of Messenger Ribonucleic Acid Encoding a Second Form of Gonadotropin-Releasing Hormone in the Rhesus Macaque Hypothalamus

A Developmental Increase in the Expression of Messenger Ribonucleic Acid Encoding a Second Form of Gonadotropin-Releasing Hormone in the Rhesus Macaque Hypothalamus

The expression, regulation and signal transduction pathways of the mammalian gonadotropin-releasing hormone receptor

Normal mammalian sexual maturation and reproductive functions require the integration and precise coordination of hormones at the hypothalamic, pituitary, and gonadal levels. Hypothalamic gonadotropin-releasing hormone (GnRH) is a key regulator in this system; after binding to its receptor (GnRHR), it stimulates de novo synthesis and release of gonadotropins in anterior pituitary gonadotropes. Since the isolation of the GnRHR cDNA, the expression of GnRHR mRNA has been detected not only in the pituitary, but also in extrapituitary tissues, including the ovary and placenta. It has been shown that change in GnRHR mRNA is one of the mechanisms for regulating the expression of the GnRHR. To help understand the molecular mechanism(s) involved in transcriptional regulation of the GnRHR gene, the 5' flanking region of the GnRHR gene has recently been isolated. Initial characterization studies have identified several DNA regions in the GnRHR 5' flanking region which are responsible for both basal expression and GnRH-mediated homologous regulation of this gene in pituitary cells. The mammalian GnRHR lacks a C-terminus and possesses a relatively short third intracellular loop; both features are important in desensitization of many others G-protein coupled receptors (GPCRs), Homologous desensitization of GnRHR has been shown to be regulated by various serine-threonine protein kinases including protein kinase A (PKA) and protein kinase C (PKC), as well as by G-protein coupled receptor kinases (GRKs). Furthermore, GnRHR was demonstrated to couple with multiple G proteins (Gq/11, Gs, and Gi), and to activate cascades that involved the PKC, PKA, and mitogen-activator protein kinases. These results suggest the diversity of GnRHR-G protein coupling and signal transduction systems. The identification of second form of GnRH (GnRH-II) in mammals adds to the complexity of the GnRH-GnRHR system. This review summaries our recent progress in understanding the regulation of GnRHR gene expression and the GnRHR signal transduction pathways.Key words: gonadotropin-releasing hormone receptor, transcriptional regulation, desensitization, signal transduction.

Molecular Cloning and Tissue-Specific Expression of a Gonadotropin-Releasing Hormone Receptor in the Japanese Eel

Gonadotropin-releasing hormone (GnRH) is a key regulatory neuropeptide involved in the control of reproduction in vertebrates. In the Japanese eel, one of the most primitive teleost species, two molecular forms of GnRH, mammalian-type GnRH and chicken-II-type GnRH (cGnRH-II), have been identified. This study has isolated a full-length cDNA for a GnRH receptor from the pituitary of the eel. The 3233-bp cDNA encodes a 380-amino acid protein which contains seven hydrophobic transmembrane domains and N- and C-terminal regions. The exon/intron organization of the open reading frame of the eel GnRH receptor gene was also determined. The open reading frame consists of three exons and two introns. The exon–intron splice site is similar to that of the GnRH receptor genes of mammals reported so far. Expression of the eel GnRH receptor was detected in various parts of the brain, pituitary, eye, olfactory epithelium, and testis. This result suggests that GnRH has local functions in these tissues in addition to its actions on gonadotropin synthesis and release in the pituitary. This tissue-specific expression pattern is similar to that of the eel cGnRH-II. Furthermore, the present eel receptor shows very high amino acid identity with the catfish and goldfish GnRH receptors, which are highly selective for the cGnRH-II. These results suggest that the cGnRH-II acts through binding to the present receptor in the eel.

Differential Distribution of Gonadotropin-Releasing Hormone-Immunoreactive Neurons in the Stingray Brain: Functional and Evolutionary Considerations

Gonadotropin-releasing hormone (GnRH) is a neuropeptide that occurs in multiple structural forms among vertebrate species. Bony fishes, amphibians, reptiles, birds, and mammals express different forms of GnRH in the forebrain and endocrine regions of the hypothalamus which regulate the release of reproductive gonadotropins from the pituitary. In contrast, previous studies on bony fishes and tetrapods have localized the chicken GnRH-II (cGnRH-II) nucleus in the midbrain tegmentum and, combined with cladistic analyses, indicate that cGnRH-II is the most conserved form throughout vertebrate evolution. However, in elasmobranch fishes, the neuroanatomical distribution of cGnRH-II and dogfish GnRH (dfGnRH) cells and their relative projections in the brain are unknown. We used high-performance liquid chromatography and radioimmunoassay to test for differential distributions of various GnRH forms in tissues from the terminal nerve (TN) ganglia, preoptic area, and midbrain of the Atlantic stingray, Dasyatis sabina. These experiments identified major peaks that coelute with cGnRH-II and dfGnRH, minor peaks that coelute with lamprey GnRH-III (lGnRH-III), and unknown forms. Immunocytochemistry experiments on brain sections show that dfGnRH-immunoreactive (-ir) cell bodies are localized in the TN ganglia, the caudal ventral telencephalon, and the preoptic area. Axons of these cells project to regions of the hypothalamus and pituitary, diencephalic centers of sensory and behavioral integration, and the midbrain. A large, discrete, bilateral column of cGnRH-II-ir neurons in the midbrain tegmentum has sparse axonal projections to the hypothalamus and regions of the pituitary but numerous projections to sensory processing centers in the midbrain and hindbrain. Immunocytochemical and chromatographic data are consistent with the presence of lGnRH-III and other GnRH forms in the TN that differ from dfGnRH and cGnRH-II. This is the first study that shows differential distribution of cGnRH-II and dfGnRH in the elasmobranch brain and supports the hypothesis of divergent function of GnRH variants related to gonadotropin control and neuromodulation of sensory function.

Biological and Immunological Characterization of Multiple GnRH in an Opisthobranch Mollusk, Aplysia californica

Gonadotropin-releasing hormone (GnRH) is a neurohormone central to the regulation of reproductive functions in vertebrates. Recently, several studies have reported the presence of GnRH immunoreactivity (IR) in a number of mollusks, suggesting that the distribution of GnRH may not be restricted to Phylum Chordata. In the present study, we extend our investigations to an opisthobranch mollusk, Aplysia californica, to characterize the source, chemical nature, and biological activity of molluscan GnRH-related molecules. Specific radioimmunoassays (RIAs) of various tissue extracts of Aplysia revealed that only ovotestis, hemocytes, and hemolymph contained significant amounts of GnRH that crossreacts with antisera raised against tunicate-I (tI) and mammalian (m) GnRH. Further RIAs and extractions revealed that the GnRH-IR in the hemolymph is biochemically and immunologically distinct from the GnRH-IR in the hemocytes and ovotestis. Using reverse-phase high-performance liquid chromatography coupled with RIAs, the GnRH-IR in the hemolymph was resolved into two major peaks. The first peak eluted earlier than most known forms of vertebrate GnRH, and the later peak coeluted with m, lamprey I, chicken II, and tI-GnRH. However, both peaks were broad and may contain a heterogeneous mixture of GnRH-IR. Immunocytochemical study showed that tI-GnRH-IR was present in the connective sheath surrounding the central nervous system, with a strong presence in what appeared to be vascular space, again suggesting the close association between Aplysia GnRH-IR and circulation. Finally, treatment of the neuroendocrine bag cells with chicken II GnRH significantly decreased the duration of the afterdischarge (AD, a characteristic pattern of electrical firing in bag cell neurons) and the number of action potentials fired during an AD, indicating the presence of a corresponding GnRH receptor in the Aplysia central nervous system. Overall, the results demonstrated the presence of multiple forms of GnRH-IR that crossreact with tI- and mGnRH antisera in A. californica and the ability of a vertebrate GnRH to alter Aplysia neural activity. Together, these data suggest that GnRH may be a factor released by the ovotestis and hemocytes into the circulation to alter neural functions. GnRH-IR produced by the latter may serve as a novel mediator of the neural and immune functions in Aplysia.

Mammalian and chicken I forms of gonadotropin-releasing hormone in the gonads of a protochordate, Ciona intestinalis.

Two forms of gonadotropin-releasing hormone (GnRH) were isolated from the gonads of the tunicate, Ciona intestinalis. The primary structure of the purified peptides was determined by MS and chemical sequence analysis. Both GnRH forms have blocked NH(2) and COOH termini, and their primary structures are identical to mammalian (mGnRH) and chicken I (cGnRH-I) forms reported previously in vertebrates. A total of 1.2 mg of purified cGnRH-I and 0.98 mg of mGnRH was obtained from 100 g of Ciona gonads. The physiological effects of native GnRHs included the induction of synthesis and secretion of sex steroids from ciona gonads and the secretion of luteinizing hormone from rat pituitary. These results suggest that the primary structure and functional roles of mGnRH and cGnRH-I have been highly conserved throughout evolution of chordates.

The expression, regulation and signal transduction pathways of the mammalian gonadotropin-releasing hormone receptor

Normal mammalian sexual maturation and reproductive functions require the integration and precise coordination of hormones at the hypothalamic, pituitary, and gonadal levels. Hypothalamic gonadotropin-releasing hormone (GnRH) is a key regulator in this system; after binding to its receptor (GnRHR), it stimulates de novo synthesis and release of gonadotropins in anterior pituitary gonadotropes. Since the isolation of the GnRHR cDNA, the expression of GnRHR mRNA has been detected not only in the pituitary, but also in extrapituitary tissues, including the ovary and placenta. It has been shown that change in GnRHR mRNA is one of the mechanisms for regulating the expression of the GnRHR. To help understand the molecular mechanism(s) involved in transcriptional regulation of the GnRHR gene, the 5' flanking region of the GnRHR gene has recently been isolated. Initial characterization studies have identified several DNA regions in the GnRHR 5' flanking region which are responsible for both basal expression and GnRH-mediated homologous regulation of this gene in pituitary cells. The mammalian GnRHR lacks a C-terminus and possesses a relatively short third intracellular loop; both features are important in desensitization of many others G-protein coupled receptors (GPCRs), Homologous desensitization of GnRHR has been shown to be regulated by various serine-threonine protein kinases including protein kinase A (PKA) and protein kinase C (PKC), as well as by G-protein coupled receptor kinases (GRKs). Furthermore, GnRHR was demonstrated to couple with multiple G proteins (Gq/11, Gs, and Gi), and to activate cascades that involved the PKC, PKA, and mitogen-activator protein kinases. These results suggest the diversity of GnRHR-G protein coupling and signal transduction systems. The identification of second form of GnRH (GnRH-II) in mammals adds to the complexity of the GnRH-GnRHR system. This review summaries our recent progress in understanding the regulation of GnRHR gene expression and the GnRHR signal transduction pathways.

Gonadotropin-releasing hormone (GnRH) neuronal systems in the pejerrey, Odontesthes bonariensis (Atheriniformes)

The brain of the pejerrey (Odontesthes bonariensis) has recently been shown to contain three forms of gonadotropin-releasing hormone (GnRH): salmon GnRH (sGnRH), chicken GnRH-II (cGnRH-II) and pejerrey GnRH (pjGnRH), nevertheless neuroanatomical studies on the distribution of these peptides are lacking. In this study we investigated the distribution of immunoreactive GnRH in the brain of adult pejerrey. Four different policlonal antisera and a monoclonal antibody against different GnRH variants were applied on cryosections and visualized using the ABC method. Three antisera (PBL#49, sGnRH#2 and cII741) revealed three different immunoreactive areas: the terminal nerve ganglion (at the junction between the olfactory bulbs and the anterior telencephalon), the preoptic area just anterior to the hypothalamus and the midbrain tegmentum. Fibers immunoreactive to GnRH were detected in different brain areas: the olfactory bulbs, the ventral thelencephalon, the hypothalamus, the mesencephalic area and an important innervation entering into the pituitary gland. Two other antibodies (LRH13 and s1668) labeled the two nuclei corresponding to the forebrain but not the midbrain tegmentum. As both antibodies have low crossreactivity to cGnRH-II, the data suggest that this group of cells express cGnRH-II. In summary, three different areas with immunoreactivity to GnRH were detected in the pejerrey brain. The distribution of sGnRH, pjGnRH and cGnRH-II expressing neurons, is discussed. Abbreviations: C‐ cerebelum ;C M ‐corpus mamilaris ;D cd ‐area dorsalis telencephali pars centralis ;D d ‐ area dorsalis telencephali pars dorsalis ;D l ‐area dorsalis telencephali pars lateralis ;D lv ‐area dorsalis telencephali pars lateralis ;D m ‐area dorsalis telencephali pars medialis ;M t ‐midbrain tegmentum; NDLl ‐ nucleus difusus lobi inferioris ;N G ‐nucleus glomerulosus ;N PGC ‐nucleus preglomerulosus pars medialis ;N PP ‐ nucleus preopticus periventricularis ;N R ‐nucleus rotundus ;N T ‐nucleus tenia; Ob ‐ olfactory bulbs; Oc ‐ optic chiasma; ON ‐ optic nerve; Otec ‐ optic tectum; Pit ‐ pituitary; Tel ‐ telencephalon; Tl ‐ torus longitudianalis; TNG ‐ terminal nerve ganglion; Vd ‐ area ventralis telencephali pars centreli ; area ventralis telencephali pars ventralis.