Chicken Gonadotropin-releasing Hormone Research Articles

Chromatographic and immunological identification of GnRH (Gonadotropin-releasing hormone) variants. Occurrence of mammalian and a salmon-like GnRH in the forebrain of an eutherian mammal: Hydrochaeris hydrochaeris (Mammalia, Rodentia).

The molecular variants of Gonadotropin releasing hormone (GnRH) in brain extracts of the eutherian mammal Hydrochaeris hydrochaeris (Mammalia, Rodentia) were characterized. An indirect method combining reverse-phase high-performance liquid chromatography (RP-HPLC) and radioimmunoassay (RIA) with different antisera was used. Two different forebrain regions (olfactory bulbs and preoptic–hypothalamic region) were analyzed. Characterization of RP-HPLC fractions from preoptic–hypothalamic extracts with three different RIA systems revealed two immunoreactive GnRH (ir-GnRH) peaks coeluting with mammalian GnRH (mGnRH) and salmon GnRH (sGnRH) synthetic standards. These results were additionally supported by serial dilution studies with specific antisera. Similar results were obtained from olfactory bulb extracts with the same methodology. However, a third ir-GnRH peak in a similar position to that of chicken GnRH II (cIIGnRH) synthetic standard was revealed. As far as we know, this is the first report showing chromatographic and immunological evidences for the presence of a second GnRH variant in the forebrain of an eutherian mammal.

An Analysis of Sex and Breeding Stage Differences in Prolactin Binding Activity in Brain and Hypothalamic GnRH Concentration in Wilson's Phalarope, a Sex Role-Reversed Species

Wilson's Phalarope (Phalaropus tricolor) is a sex role-reversed species in which incubation of eggs and care of young is performed exclusively by the male. Plasma levels of prolactin (PRL), the hormone most associated with parental care in birds, are higher in incubating males than in nonincubating males or females. Conversely, plasma testosterone levels are reduced in males during incubation. In an attempt to characterize the physiological basis of this unusual parental care system, we used quantitative film autoradiography and densitometry to measure the specific bindingin vitroof125I-ovine PRL to 12 brain regions in females, nonincubating males, and incubating males during the normal breeding season. We also measured hypothalamic chicken gonadotropin-releasing hormone I (cGnRH-I) in three brain areas in these same birds, as well as plasma levels of PRL and testosterone. Analysis revealed that cGnRH-I concentrations in the preoptic area and plasma testosterone levels were significantly lower in incubating males than in nonincubating males. Specific binding of125I-ovine PRL was detected in choroid plexus and in several diencephalic brain regions of both sexes, with highest binding activity recorded in the dorsolateral thalamus, medial habenula, nucleus subrotundus, and preoptic area. When adjustments were made for the large number of comparisons performed, specific binding did not vary significantly by sex or breeding stage in any single brain region. However, average specific binding values in nonincubating males exceeded those of incubating males in 9 of the 11 PRL-sensitive regions examined. Increased occupancy of the receptor by endogenous PRL during incubation could have contributed to this result, since plasma PRL levels were elevated in incubating males. In addition, PRL binding activity in several of these brain regions tended to correlate negatively with plasma PRL. The two exceptions to this general pattern were the preoptic area and the lateral septum, where mean specific binding was 14–15% higher in incubating males than in nonincubating males. This raises the interesting possibility that PRL sensitivity is up-regulated during incubation in some regions of the male phalarope brain, such as the preoptic area and lateral septum, that have been implicated in PRL-modulated changes in behavior and reproductive activity during this breeding stage.

Effects of thyroxine (T4) or triiodothyronine (T3) replacement therapy on the programming of seasonal reproduction and postnuptial molt in thyroidectomized male American tree sparrows (Spizella arborea) exposed to long days.

This study tested the hypothesis that T3 (triiodothyronine) is the tissue-active "seasonality" hormone by determining whether T3 could mimic T4 (thyroxine) and program photostimulated thyroidectomized (THX) male American tree sparrows (Spizella arborea) for three components of seasonality (i.e., full-blown testicular growth, photorefractoriness, and postnuptial molt). Photosensitive males were radiothyroidectomized, transferred to long days 4 weeks later, and administered 14 daily injections (s.c.) of alkaline saline (V) containing 0.1, 1, or 10 micrograms T4 or T3. THX and thyroid-intact (THI) controls received only V. After 5 additional weeks on long days, all birds were tested for photosensitivity/photorefractoriness. Periodically during the experiment, primary flight feathers were scored for molt, and testis length was monitored by laparotomy. As an independent measure of reproductive (i.e., photosensitive vs. photorefractory) state, hypothalami collected at the end of the experiment were assayed for cGnRH-I (chicken gonadotropin-releasing hormone I) content. Like THI controls, THX males administered 1 or 10 micrograms T4 exhibited full-blown testicular growth and then regression, initiated molt, and had low hypothalamic cGnRH-I, indicating that photostimulated birds that received mid- or high-dose T4 replacement therapy had been programmed for all three components of seasonality. On the other hand, both THX controls and THX males administered low-dose (0.1 microgram) T3 replacement therapy exhibited only modest testicular growth, signifying that neither group had been programmed for any component of seasonality. By contrast, photostimulated THX males that received 0.1 microgram T4, or 1 or 10 micrograms T3, were programmed for testicular growth, but not for photorefractoriness or molt. Collectively, these results show that subcutaneously administered T3 mimicked T4 imperfectly and suggest either that T3 does not program photostimulated male tree sparrows for photorefractoriness and postnuptial molt, or that T3 does not cross the blood-brain barrier as efficiently as does T4.

Characterization of Two New preproGnRH mRNAs in the Tree Shrew: First Direct Evidence for Mesencephalic GnRH Gene Expression in a Placental Mammal

Reproductive maturation and regulation is centrally orchestrated by gonadotropin-releasing hormone (GnRH). GnRH produced in the vertebrate hypothalamus acts on the pituitary to regulate gonadotropins. In nonplacental mammalian species, it has recently been shown that a second GnRH gene is expressed in mesencephalic cells. Here, we report the cDNA sequences and expression patterns for two distinct genes encoding the hypothalamic and mesencephalic GnRH forms in the brain of a placental mammal, the tree shrew (Tupaia glis belangeri). The novel mammalian GnRH form, designated here as [His5Trp7Tyr8]GnRH (often called chicken GnRH II), is expressed in neurons of the mesencephalon and is the first nonhypothalamic form to be isolated from a mammal. Its peptide sequence is identical to the form previously reported in fish, amphibians, reptiles, and birds, revealing that it has remained unchanged for 500 million years. In contrast, the sequences of the hypothalamic GnRH decapeptides vary by as much as 50% across vertebrate species. The remarkable sequence conservation of mesencephalic GnRH suggests that it has been highly constrained throughout evolution, perhaps indicating an important, conserved nongonadotropic role. The discovery and localization of two mRNAs encoding distinct GnRH forms in an advanced mammal suggest that other mammals, including primates, may also have a second GnRH gene with expression localized in the midbrain.

The Thyroid and the Hypothalamus–Pituitary–Ovarian Axis in American Tree Sparrows (Spizella arborea)

Prompted by evidence of a linkage between the thyroid gland and seasonal reproduction in obligately photoperiodic American tree sparrows (Spizella arborea), we measured circulating T4(L-thyroxine), hypothalamic cGnRH-I (chicken gonadotropin-releasing hormone, variant I), pituitary and plasma LH (luteinizing hormone), and ovarian mass in euthyroid females moved from short to long days. Our purpose was to correlate temporal changes in T4and cGnRH-I with each other and with pituitary and plasma LH and ovarian mass. T4increased early during photostimulation and peaked at Week 1 or 2 on 20L:4D (20 hr light:4 hr dark per day). Hypothalamic cGnRH-I also increased during photostimulation and peaked between Weeks 2 and 4. Pituitary and plasma LH peaked at Weeks 4 and 1, respectively, and ovarian mass increased significantly by Week 6. In a separate study, we charted the progression of postnuptial molt. Molt of the primary remiges began at Week 9, proceeded rapidly through Week 15, and was complete by Week 18. In view of a demonstrated role for the thyroid gland in the control of seasonal reproduction in female tree sparrows and the time-dependent effects of thyroidectomy after photostimulation in male tree sparrows, the observation that circulating T4increases early during photostimulation suggests the possibility of a causal relationship between the thyroid gland and the hypothalamus–pituitary–ovarian axis, hence athyroid–hypothalamus–pituitary–ovarian axis.

The Timing of Thyroid-Dependent Programming in Seasonally Breeding Male American Tree Sparrows (Spizella arborea)

There is convincing evidence that euthyroid male American tree sparrows are already programmed for seasonal reproduction and postnuptial molt by Week 4 of photostimulation. To explore more precisely when, during early photostimulation, thyroid-dependent programming of seasonal events occurs, photosensitive male tree sparrows were radiothyroidectomized or sham thyroidectomized on the first day of photostimulation (Week 0) or at Weeks 1 or 3 thereafter. Birds were monitored for testicular growth and regression over 8 or 12 weeks and then tested for photosensitivity or photorefractoriness by exposing them to constant light and exogenousL-thyroxine for 4 weeks. Molt of the primary flight feathers was scored periodically, and at the end of the experiment, hypothalami were saved for cGnRH-I (chicken gonadotropin-releasing hormone I) assay. Because (1) the thyroid is already dysfunctional by Day 4 after radiothyroidectomy, (2) thyroid-dependent photoinduced gonadal growth is programmed growth, and as demonstrated here, (3) testis length at Week 6 on long days (i.e., maximum or near-maximum testis size) is independent of the time of thyroidectomy at or after the onset of photostimulation, we conclude that male American tree sparrows were programmed for photoperiodic testicular growth during the first week of photostimulation. Based on measurements of testis length and hypothalamic cGnRH-I content, only 2 of 11 birds thyroidectomized at Week 1 were already programmed for photorefractoriness by Week 1, whereas all birds thyroidectomized at Week 3 were so programmed by Week 3. Marked differences in molt scores between photorefractory (thyroidectomized and euthyroid) and photosensitive (thyroidectomized) birds argue that postnuptial molt also was programmed roughly between Weeks 1 and 3. To explain these results, we ascribe organizational-like actions directly or indirectly to endogenous thyroid hormones and argue that the onset of photostimulation opens a window, during which time euthyroid male American tree sparrows are programmed for seasonality. A clear dissociation of photorefractoriness from photoperiodic testicular growth in birds thyroidectomized at Week 1 suggests that separate control circuits mediate these two annually periodic events.

Characterization and localization of gonadotropin-releasing hormone in the brain and pituitary of the three-spined stickleback, Gasterosteus aculeatus

Radioimmunoassay (RIA) studies on highperformance liquid chromatography (HPLC) fractions of brain extracts of the three-spined stickleback, Gasterosteus aculeatus, provided evidence for at least two forms of gonadotropin-releasing hormone (GnRH). One form showed chromatographic and immunological properties similar to that of synthetic salmon GnRH (sGnRH). A second, unidentified form of GnRH eluted in the same position as chicken GnRH I (cGnRH-I); however, it did not cross-react in a cGnRH-I RIA. Furthermore, it cannot be excluded that chicken GnRH II (cGnRH-II) and maybe one other unidentified form are present in the stickleback. The distribution of GnRH in the brain of breeding adult male sticklebacks was studied by use of immunohistochemistry. Two antisera against sGnRH and antisera against mGnRH and cGnRH-II were applied on cryosections and visualized using the peroxidase-antiperoxidase method. Staining patterns were similar after incubations with all four antisera. Immunoreactive fibers were found in most parts of the brain. Three distinct groups of GnRH-immunoreactive perikarya were found in the nucleus olfactoretinalis, in the nucleus anterior periventricularis, and in the nucleus lateralis tuberis. Moreover, weakly stained cells occurred in a periventricular position in the midbrain. The proximal pars distalis of the pituitary, housing the gonadotropic cells, was richly innervated by GnRH-positive fibers. In the pars intermedia and in the rostral pars distalis, immunoreactive fibers were absent.

Evolutionary aspects of gonadotropin-releasing hormone and its receptor.

1. Gonadotropin-releasing hormone (GnRH) was originally isolated as a hypothalamic peptide hormone that regulates the reproductive system by stimulating the release of gonadotropins from the anterior pituitary. However, during evolution the peptide was subject to gene duplication and structural changes, and multiple molecular forms have evolved. 2. Eight variants of GnRH are known, and at least two different forms are expressed in species from all vertebrate classes: chicken GnRH II and a second, unique, GnRH isoform. 3. The peptide has been recruited during evolution for diverse regulatory functions: as a neurotransmitter in the central and sympathetic nervous systems, as a paracrine regulator in the gonads and placenta, and as an autocrine regulator in tumor cells. 4. Evidence suggests that in most species the early-evolved and highly conserved chicken GnRH II has a neurotransmitter function, while the second form, which varies across classes, has a physiologic role in regulating gonadotropin release. 5. We review here evolutionary aspects of the family of GnRH peptides and their receptors.

Concentration of chicken gonadotropin-releasing hormones I and II in microdissected areas of turkey hen brain during the reproductive cycle

Chicken gonadotropin-releasing hormones I and II (cGnRH I and II) were measured by radioimmunoassay (RIA) in extracts of microdissected regions of turkey hen brain (preoptic area [POA], region of periventricular nuclei [PHN], septum [SEP], hippocampus [HP], dorsomedial thalamus/habenula [DMT], midbrain central gray [MCG], and caudal lateral hypothalamus [LH]) at five stages of the reproductive cycle: before photostimulation, during egg laying, during incubation, during photorefractoriness, and after return to short daylengths. The highest concentration of cGnRH I occurred in PHN, followed by POA, SEP, DMT, HP, LH, and MCG, in decreasing order, whereas the highest concentration of cGnRH II occurred in SEP, followed by POA, DMT, HP, MCG, PHN, LH. These results agree, with some exceptions, with the distribution of fibers and cells as determined by immunohistochemistry. cGnRH II was from 1.3 to 24 times as abundant as cGnRH I in different brain areas. During incubation, cGnRH I concentrations were significantly elevated in the POA and cGnRH II levels were significantly elevated in HP; few other significant differences were detected. Correlation analysis detected occasional significant positive and negative correlations between cGnRH I and II concentrations in forebrain areas and MCG of laying birds and in PHN and LH of incubating birds. These results demonstrate an approximate correspondence between hormone concentrations measured in tissue extracts by RIA and immunohistochemistry and indicate an abundance of cGnRH II as compared with cGnRH I. cGnRH I and II concentrations did not, however, change in parallel in all brain areas, suggesting that these peptides do not function in an exactly parallel fashion. Thus, the extent to which cGnRH II is involved in gonadotropin release remains unresolved.

Identification of the Molecular Forms of and Steroid Hormone Response to Gonadotropin-Releasing Hormone in the Australian Lungfish, Neoceratodus forsteri

Gonadotropin-releasing hormone (GnRH) peptides in the hypothalamus of the lungfish, Neoceratodus forsteri, were investigated by reverse-phase HPLC and RIA with region-specific antisera. Both chicken GnRH II and mammalian GnRH were identified, the latter being present in greater concentration. The steroidogenic response to a single intracardiac injection of synthetic mammalian GnRH was investigated in early and late spring (beginning and end of spawning season) and in early autumn. In early spring, both sexes responded with rapid and transient elevation of circulating steroid hormones. Testosterone in males showed the greatest response by elevating from 120 to 240 nmol/liter within 2 min of the injection and returning to ∼100 nmol/liter within 15 min. Female lungfish showed a similar but slightly less dramatic response in circulating estradiol and testosterone. The responses of both males and females were reduced in late spring and abolished in early autumn, which is indicative of a period of seasonal refractoriness.

Immunocytochemical localization of mammalian GnRH (gonadotropin-releasing hormone) and chicken GnRH-II in the brain of the European silver eel ( Anguilla anguilla L.)

Using specific antibodies for the two molecular forms of gonadotropin-releasing hormone (GnRH) present in the European eel, Anguilla anguilla, (mammalian GnRH, mGnRH, and chicken GnRH II, cGnRH-II), we employed immunocytochemistry to determine the distribution of these two peptides in the brain and in the pituitary. The results indicate that mGnRH and cGnRH-II are localized in different neurons: mGnRH-immunoreactive (ir) perikaria were observed in the olfactory bulbs, the junction between olfactory bulbs and telencephalon (nucleus olfactoretinalis), the telencephalon, the preoptic region and the mediobasal hypothalamus. These cell bodies are located along a continuum of ir-fibers that could be traced from the olfactory nerve to the pituitary. Mammalian GnRH-ir fibers were detected in many parts of the brain (olfactory bulbs, ventral telencephalon, hypothalamus, optic tectum, mesencephalon) and in the pituitary. Chicken GnRH-II-ir cell bodies were detected in the nucleus of the medial longitudinal fasciculus of the midbrain tegmentum, but only scattered fibers could be detected in different parts of the brain. The pituitary exhibited very few cGnRH-II-ir fibers, contrasting with an extensive mGnRH innervation. These results are in agreement with our previous data obtained in the same species using specific radioimmunoassays for mGnRH and cGnRH-II. They demonstrate a differential distribution of the two forms of GnRH in the brain of the eel, as in the brain of some other vertebrate species, and suggest differential physiological roles for the two GnRH forms in the eel. They also provide information concerning the evolution of the GnRH systems in vertebrates.

Gonadotropin releasing hormone (GnRH) neurons project to growth hormone and somatolactin cells in the steelhead trout

Analysis of gene expression using gonadotropin-releasing hormone (GnRH) antisense oligonucleotide confirmed by immunocytochemical localization the occurrence of GnRH neurons along the nervus terminalis in the steelhead trout (Oncorhynchus mykiss). Double-label immunocytochemistry revealed the distribution of mammalian (m), salmon (s) and chicken II (cII)-type GnRHs and various pituitary hormones. Both sGnRH and mGnRH appeared to be colocalized in the same cells of the nervus terminalis. Chicken GnRH II-immunoreactivity was found only in fibers and terminals. In the younger fish [73 and 186 days after fertilization (DAF)] GnRH neurons were seen rostral to the olfactory bulb. A novel GnRH ganglion, along the nervus terminalis, was found at the cribiform bone (gCB). A few non-immunoreactive rounded cells were seen among the GnRH neurons. A second smaller ganglion was seen at the most rostrally located part of the ventromedial olfactory bulb (gROB). In the older fish (850 DAF) GnRH neurons were also observed in the basal forebrain. A small group of neurons (2-3 cells), at the caudoventromedial border of the olfactory bulb, formed the ganglion terminale. Occasionally isolated GnRH-immunoreactive cells were seen at the base of the olfactory epithelium, along the ventromedial margins of the olfactory nerve. GnRH-immunoreactive and GnRH mRNA expressing neurons were absent from midbrain regions at the ages observed. GnRH-immunoreactive fibers were present only in older fish. The pattern of distribution of fibers that were immunoreactive to all three forms of GnRH was identical. Fibers were seen along the medial side of the olfactory nerve, throughout the brain and in the pituitary, associated with growth hormone and somatolactin cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Insulin and Insulin-like Growth Factors I and II Induce Final Maturation of Oocytes of Red Seabream, Pagrus major, in Vitro

The effects of various growth factors and peptides on in vitro germinal vesicle breakdown (GVBD) were examined in oocytes of the red seabream, Pagrus major , a daily spawning marine teleost. The oocytes used underwent GVBD in response to HCG but not to 17α,20β-dihydroxy-4-pregnen-3-one (DHP). Insulin (1 μ M) and insulin-like growth factor-1 (IGF-I, 10 n M) and IGF-II (13 n M) induced GVBD. Salmon gonadotropin-releasing hormone (GnRH), chicken GnRH (100 n M), angiotensin II and III (10 μg/ml), inhibin A, activin A (200 ng/ml), and epidermal growth factor (1 n M) were ineffective. IGF-I was the most potent inducer of GVBD. In the presence of DHP, oocytes underwent GVBD in response to low doses of IGF-I or -II or insulin which alone were ineffective. GVBD occurred in response to DHP after oocytes were preincubated with IGF-I or with HCG. Actinomycin D inhibited GVBD induced by HCG alone or in combination with DHP, but actinomycin D did not block GVBD induced by IGF-I alone or in combination with DHP. Cycloheximide totally inhibited induction of GVBD by HCG or IGF-I, alone or in combination with DHP. These results suggest, for the first time in teleost, that growth factors, such as IGF-I, are involved in the induction of GVBD of oocytes. IGF-I appears to act directly on oocytes, and not via maturation-inducing hormone production, to induce GVBD.

Antibodies against different forms of GnRH distinguish different populations of cells and axonal pathways in a urodele amphibian, Taricha granulosa

Neurons immunoreactive to the peptide hormone gonadotropin-releasing hormone (GnRH) have been identified in the posterior diencephalon or anterior midbrain of diverse vertebrates. These cells are located caudal to the more well-characterized GnRH neurons in the nervus terminalis and septo-preoptic area, and are believed to express one or more of the nonmammalian forms of the GnRH. The present study utilized immunocytochemical techniques to determine whether the posterior GnRH group is present in a urodele amphibian, the newt Taricha granulosa. Antibodies directed against different molecular forms of GnRH were used to evaluate the immunological properties of GnRH-containing neurons in amphibians. An antibody selective for mammalian GnRH labeled perikarya in the nervus terminalis (terminal nerve) and septo-preoptic region, as described previously. Thick fibers that arise from terminal nerve and septo-preoptic neurons project mainly to the median eminence, medial pallium and habenula. An antibody selective for chicken GnRH II labeled cell bodies in the paraventricular organ and posterior tubercle of the caudal diencephalon, and thin fibers that project widely throughout the central nervous system. Region-specific staining with different GnRH antibodies supports the interpretation that different molecular forms of GnRH are expressed by neuroanatomically distinguishable systems.

Isolation, characterization and expression of cDNAs encoding the catfish‐type and chicken‐II‐type gonadotropin‐releasing‐hormone precursors in the African catfish

The cDNAs encoding the catfish prepro-gonadotropin-releasing hormone and the chicken prepro-gonadotropin-releasing hormone II of the African catfish (Clarias gariepinus) have been isolated and sequenced. The catfish gonadotropin-releasing-hormone precursor and the chicken gonadotropin-releasing-hormone-II precursor have the same overall architecture as other gonadotropin-releasing-hormone precursors identified so far; each is composed of a signal peptide, gonadotropin-releasing hormone and a gonadotropin-releasing-hormone-associated peptide which is connected to gonadotropin-releasing hormone and chicken gonadotropin-releasing hormone II, in combination with the Gly-Lys-Arg sequence, are highly conserved during evolution when compared with the corresponding regions of mammalian, avian (chicken gonadotropin-releasing hormone I) and other fish gonadotropin-releasing-hormone precursors. However, the gonadotropin-releasing-hormone-associated peptide regions are markedly divergent. Northern-blot analysis revealed the presence of a single catfish gonadotropin-releasing-hormone mRNA species of about 470 bases, and the presence of a single chicken gonadotropin-releasing-hormone-II mRNA species of about 650 bases in the African catfish brain. In situ hybridization revealed catfish gonadotropin-releasing-hormone cell bodies rostro-caudally scattered in the olfactory nerve, along both sides of the midline of the telencephalon, in the preoptic area of the ventral hypothalamus, and in the infundibular stalk close to the pituitary. Chicken gonadotropin-releasing-hormone-II cell bodies, however, were exclusively found in the midbrain tegmentum.

Functional and Molecular Aspects of the Gonadotropin Releasing Hormones in the African Catfish, Clarias Gariepinus

Two molecular forms of GnRH were identified in the African catfish, Clarias gariepinus: catfish gonadotropin-releasing hormone (cfGnRH) and chicken gonadotropin-releasing hormone (cGnRH-II). They differ in gonadotropine hormone (GTH)-release activity, cGnRH-II being about a hundred times as potent as cfGnRH. This difference corrcsponds to different GnRH receptor binding affinities. cDNAs encoding both GnRH preprohormones were cloned and specific antibodies were raised against the GnRHs. By immunocytochemistry and in situ hybridization cGnRH-II was localized in the midbrain tegmentum, cfGnRH in the rostro-ventral forebrain.

Brain Content of cGnRH I and II during Embryonic Development in Chickens

We used specific radioimmunoassays to measure chicken gonadotropin-releasing hormones I and II (cGnRH I and II) in extracts of chicken brain as a first step in determining whether these peptides may function identically during embryonic development in birds. In three experiments chicken embryo brains were removed at various times between Day 6 of incubation and hatching. In Experiment 3, the sex of embryos was identified by means of polymerase chain reaction amplification of a W chromosome-specific DNA sequence. Brain concentrations of cGnRH I increased sharply from Day 6 of incubation to Day 8, then decreased until Day 10 to Day 12, followed by an increase beginning on approximately Day 17 and continuing until hatch. In contrast, cGnRH II concentration remained low until about Day 14 of incubation, then increased progressively until, at hatch, brain content of cGnRH II was approximately 9 to 11 times that of cGnRH I. The difference in development pattern and total content of these peptides supports the view that any physiologic function they may have might be differentiated as early as Day 7 of incubation.

Chicken Gonadotropin-Releasing Hormones Enhance Soluble and Insoluble Fibronectin Production by Granulosa Cells of the Domestic Fowl In Vitro

Experiments were conducted in vitro to examine the effect of two chicken gonadotropin-releasing hormones, cGnRH-I ([Gln8]-GnRH) and cGnRH-II ([His5,Trp7,Tyr8]-GnRH), on fibronectin (soluble and insoluble) production by chicken granulosa cells isolated from the largest (F1; about 35 mm in diameter), and third largest (F3; 15 to 20 mm in diameter) preovulatory follicles as well as from a pool of immature small yellow follicles (SYF; 6 to 8 mm in diameter). The amounts of soluble fibronectin (fibronectin secreted into the incubation medium) and insoluble fibronectin (fibronectin associated with cells plus fibronectin attached to culture substratum) were quantified with a specific ELISA. Fibronectin secreted into the incubation medium (soluble fibronectin) by unstimulated cells increased with advanced stages of follicular maturation. Addition of both cGnRH-I and -II increased the amount of fibronectin secreted into the incubation medium by all follicular cell types. The amount of insoluble fibronectin in culture wells that contained unstimulated cells also increased with advanced stages of follicle development. Both cGnRH-I and -II increased the quantity of insoluble fibronectin by granulosa cells from all follicle types. Total (soluble plus insoluble) fibronectin production was elevated when cGnRH-I or -II was added to F1, F3, and SYF granulosa cells. The magnitude of cGnRH-I or -II stimulation (percentage increase) of soluble, insoluble, or total fibronectin production was calculated as a multiple of the unstimulated (control) value for each follicle type, and they were greatest in cells derived from developing and immature follicles. These results indicate that homologous cGnRH-I and -II are capable of directly modulating the physiology of the avian ovary.

Immunohistochemical localization of chicken gonadotropin‐releasing hormones I and II (cGnRH I and II) in turkey hen brain

The distribution of cells and fibers immunoreactive (ir) for either chicken gonadotropin-releasing hormone I (cGnRH I; [Gln8]GnRH) or II ([His5,Trp7,Tyr8]GnRH) was determined in brains of turkey hens to reveal whether these peptides occur in separate neuronal systems. ir-cGnRH I cells were located: along the medial aspect of the ventriculus lateralis, nucleus accumbens, and bed nucleus of the stria terminalis; ventral to the tractus septomesencephalicus and extending medially to the third ventricle, and caudally into the lateral hypothalamic area; and in a diffuse band extending from the nucleus preopticus medialis to the nucleus dorsomedialis anterior thalami. cGnRH I fibers were evident in these areas in addition to the hippocampus, nucleus subhabenularis medialis, nucleus ventromedialis hypothalami, and median eminence. Two groups of ir-cGnRH II cells were observed: a magnocellular group lying between the substantia grisea centralis and the nucleus ruber; and a parvicellular group lying medial to the nucleus of the basal optic root and extending into the lateral hypothalamic area. ir-cGnRH II fibers were prominent in limbic structures (cortex piriformis, lateral to nucleus taeniae, hippocampus); olfactory areas (tuberculum olfactorium, nucleus subhabenularis lateralis, nucleus septalis lateralis); areas that in other avian species have steroid-concentrating cells or receptors (medial edge of lobus parolfactorius, nucleus septalis medialis, nucleus periventricularis magnocellularis, nucleus dorsomedialis posterior thalami); and areas containing ir-GnRH I cells or fibers but not in median eminence. These results suggest that cGnRH I and II occur in separate neuronal systems and that cGnRH II does not directly promote pituitary gonadotropin secretion.

Precocious puberty in tamoxifen‐treated cockerels: Hypothalamic gonadotrophin‐releasing hormone‐I and plasma luteinising hormone, prolactin, growth hormone and testosterone

1. The administration of the anti-oestrogen, tamoxifen (TAM) to juvenile chicks results in precocious puberty. In the present study the effects of TAM administration (1 mg/kg body weight on alternate days from 12 d of age) on testicular function, hypothalamic chicken gonadotropin-releasing hormone (cGn-RH-I), plasma luteinising hormone (LH), growth hormone (GH), prolactin (PRL) and testosterone were studied in juvenile White Leghorn cockerels. 2. The increase in hypothalamic GnRH-I content which occurs during sexual development was advanced in TAM-treated birds, in association with precocious testicular development, an early rise of plasma testosterone content and enhanced comb growth. 3. Plasma LH concentrations behaved similarly and were higher in the TAM-treated than in control birds, during most of the experimental period. Plasma PRL concentration, which is high at hatching, decreased more quickly in TAM-treated than in control birds; plasma GH values were not consistently affected by TAM treatment. 4. Both the growth and the involution of the bursa of Fabricius in the TAM-treated cockerels preceded that in the control chicks. 5. It is concluded that TAM treatment induces precocious puberty in the cockerel by blocking the negative feedback action of aromatised testicular androgens on the hypothalamus.