Reproduction and beyond,kisspeptin in ruminants

Kisspeptin(Kp) is synthesized in the arcuate nucleus and preoptic area of the hypothalamus and is a regulator of gonadotropin releasing hormone in the hypothalamus.In addition,Kp may regulate additional functions such as increased neuropeptide Y gene expression and reduced proopiomelanocortin(POMC) gene expression in sheep.Other studies have found a role for Kp to release growth hormone(GH),prolactin and luteinizing hormone(LH)from cattle,rat and monkey pituitary cells.Intravenous injection of Kp stimulated release LH,GH,prolactin and follicle stimulating hormone in some experiments in cattle and sheep,but other studies have failed to find an effect of peripheral injection of Kp on GH release.Recent studies indicate that Kp can stimulate GH release after intracerebroventricular injection in sheep at doses that do not release GH after intravenous injection.These studies suggest that Kp may have a role in regulation of both reproduction and metabolism in sheep.Since GH plays a role in luteal development,it is tempting to speculate that the ability of Kp to release GH and LH is related to normal control of reproduction.     

Comparative Aspects of the Endotoxin- and Cytokine-Induced Endocrine Cascade Influencing Neuroendocrine Control of Growth and Reproduction in Farm Animals

Disease in animals is a well-known inhibitor of growth and reproduction. Earlier studies were initiated to determine the effects of endotoxin on pituitary hormone secretion. These studies found that in sheep, growth hormone (GH) concentration was elevated, whereas insulin-like growth factor-I (IGF-I) was inhibited, as was luteinizing hormone (LH). Examination of the site of action of endotoxin in sheep determined that somatotropes expressed the endotoxin receptor (CD14) and that both endotoxin and interleukin-Iβ activated GH secretion directly from the pituitary. In the face of elevated GH, there is a reduction of IGF-I in all species examined. As GH cannot activate IGF-I release during disease, there appears to be a downregulation of GH signalling at the liver, perhaps related to altered nitration of Janus kinase (JAK). In contrast to GH downregulation, LH release is inhibited at the level of the hypothalamus. New insights have been gained in determining the mechanisms by which disease perturbs growth and reproduction, particularly with regard to nitration of critical control pathways, with this perhaps serving as a novel mechanism central to lipopolysaccharide suppression of all signalling pathways. This pathway-based analysis is critical to the developing novel strategies to reverse the detrimental effect of disease on animal production.     

Regulation of the growth hormone and luteinizing hormone response to endotoxin in sheep

Infectious disease processes cause physiological adaptations in animals to reorder nutrient partitioning and other functions to support host survival. Endocrine, immune and nervous systems largely mediate this process. Using endotoxin injection as a model for catabolic disease processes (such as bacterial septicemia), we have focused our attention on regulation of growth hormone (GH) and luteinizing hormone (LH) secretion in sheep. Endotoxin produces an increase in plasma GH and a decrease in plasma LH concentrations. This pattern can be reproduced, in part, by administration of various cytokines. Antagonists to both interleukin-1 (IL-1) and tumor necrosis factor (TNF) given intravenously (IV) prevented the endotoxin-stimulated increase in GH. Since endotoxin will directly stimulate GH and LH release from cultured pituitary cells, the data suggest a pituitary site of action of the endotoxin to regulate GH. Studies with portal vein cannulated sheep indicated that gonadotropin releasing hormone was inhibited by endotoxin, suggesting a central site of action of endotoxin to regulate LH. However, other studies suggest that endotoxin may also regulate LH secretion at the pituitary. Thus, IL-1 and TNF regulate GH release from the pituitary gland while endotoxin induces a central inhibition of LH release.     

Effects of intracerebroventricular injections of leptin on the release of luteinizing hormone and growth hormone in castrated calves

The purpose of the present study was to clarify the hypothalamic action of leptin on the secretion of luteinizing hormone (LH) and growth hormone (GH) in cattle. Intracerebroventricular (the third ventricle) injections of leptin were given to fully fed castrated Holstein calves. Blood samples were collected at 10‐min intervals for 60 min after injection and 20‐min intervals for 60 min before injection and for 60–180 min after injection through an indwelling catheter in the external jugular vein. Plasma LH and GH levels were examined by homologous radioimmunoassay. The administration of 10 µg of leptin stimulated a significant (P < 0.05) release of GH but not LH. Average GH levels began to rise after 30 min and were significantly increased at 40, 50 and 60 min after the injection, compared with the respective control values (P < 0.05). The present result suggests that leptin may act partly on the hypothalamus to stimulate the release of GH in castrated calves.     

Effects of serotonin injected into the third ventricle on prolactin and growth hormone secretion in Holstein steers

In order to clarify the role of serotonin (5-HT) in the regulation of pituitary hormones, the effects of 5-HT injected into the third ventricle (3V) on prolactin (PRL) and growth hormone (GH) release were investigated in Holstein steers. A chronic cannula was implanted in 3V by stereotaxic surgery under general anesthesia. After sufficient recovery from surgery, 5-HT (0, 0.1, 1.0, 2.0 mg) was injected into via the cannula and blood samples were collected over 4 h. Plasma PRL and GH concentrations were determined by radioimmunoassay. PRL release was significantly stimulated by the injection of 5-HT. The increase in PRL was observed at 20 min after the injection at three doses and the highest dose (2.0 mg) was the most effective in stimulating PRL release. The injection of 5-HT into 3V, at all doses tested, did not alter GH release significantly. Our results suggest that 5-HT is involved in the regulation of PRL release partly through the hypothalamus in cattle.     

Kisspeptins and the reproductive axis: potential applications to manage reproduction in farm animals

Kisspeptins (Kp) are a family of neuropeptides produced mainly by two hypothalamic neuronal cell populations. They have recently emerged as a major regulator of the gonadotropin axis and their action is located upstream of the gonadotropin-releasing hormone (GnRH) cell population. In less than 10 yr a growing body of literature has demonstrated the involvement of these peptides in most, if not all, aspects of reproductive axis maturation and function. In contrast to these abundant basic research studies, few experiments have evaluated the potential application of Kp as tools to manipulate reproduction in domestic animals. In mammals, exogenous Kp administration potently stimulates gonadotropin secretion. This action is exerted mainly, if not exclusively, through the stimulation of GnRH release. Intravenous, intraperitoneal, or subcutaneous administration of Kp induced a robust and rapid increase in plasma gonadotropins (luteinizing hormone [LH] and follicle-stimulating hormone [FSH]). However, this stimulatory effect is of short duration. Prolonged LH and FSH release over several hours can be achieved only when Kp are given as repeated multiple bolus or as an infusion. Kp administration was used in two experimental models, ewe and pony mare, with the aim of inducing well-timed and synchronized ovulations. During the breeding season, progesterone-synchronized ewes were given an intravenous infusion of Kp starting 30 h after the removal of progesterone implants. An LH surge was induced in all Kp-treated animals within 2 h of infusion onset. In contrast, in pony mares a constant infusion of Kp for 3 d in the the late follicular phase was unable to induce synchronized ovulation. Another set of studies showed that Kp could be used to activate reproductive function in acyclic animals. Pulsatile administration of Kp in prepubertal ewe lambs was shown to activate ovarian function, leading to enhanced ovarian steroidogenesis, stimulation of LH preovulatory surge, and ovulation. In anestrous ewes, an intravenous infusion of a low dose of Kp induced an immediate and sustained release of gonadotropins, followed a few hours later by an LH surge. This hormonal pattern mimicked hormonal changes normally observed during the estrous cycle follicular phase and was associated with a high percentage of ovulating animals (80%). In summary, exogenous administration of Kp appears to be a new tool to manipulate reproduction. However, optimal doses and periods of treatment should be defined for each species, and the development of powerful analogs or long-term release formulations is necessary before large-scale applications in domestic animals could be envisaged.     

Effect of LPS on Reproductive System at the Level of the Pituitary of Anestrous Ewes

In our research we focused our attention on the effect of the immune stress induced by bacterial endotoxin–lipopolysaccharide (LPS) on the hypothalamic–pituitary–gonadal axis (HPG) at the pituitary level. We examined the effect of intravenous (i.v.) LPS injection on luteinizing hormone (LH) and follicle‐stimulating hormone (FSH) release from the anterior pituitary gland (AP) in anestrous ewes. The effect of endotoxin on prolactin and cortisol circulating levels was also determined. We also researched the effect of immune challenge on the previously mentioned pituitary hormones and their receptors genes expression in the AP. Our results demonstrate that i.v. LPS injection decreased the plasma concentration of LH (23%; p < 0.05) and stimulates cortisol (245%; p < 0.05) and prolactin (60%; p < 0.05) release but has no significant effect on the FSH release assayed during 6 h after LPS treatment in comparison with the control levels. The LPS administration affected the genes expression of gonadotropins’β‐subunits, prolactin and their receptors in the AP. Endotoxin injection significantly decreased the LHβ and LH receptor (LHR) gene expression (60%, 64%; p < 0.01 respectively), increased the amount of mRNA encoding FSHβ, FSH receptor (FSHR) (124%, 0.05; 166%, p < 0.01; respectively), prolactin and prolactin receptor (PRLR) (50%, 47%, p < 0.01; respectively). The presented, results suggest that immune stress is a powerful modulator of the HPG axis at the pituitary level. The changes in LH secretion could be an effect of the processes occurring in the hypothalamus. However, the direct effect of immune mediators, prolactin, cortisol and other components of the hypothalamic pituitary–adrenal (HPA) axis on the activity of gonadotropes has to be considered as well. Those molecules could affect LH synthesis directly through a modulation at all stages of LHβ secretion as well as indirectly influencing the GnRHR expression and leading to reduced pituitary responsiveness to GnRH stimulation.     

Endotoxin inhibition of luteinizing hormone in sheep

Administration of endotoxin suppresses circulating concentration of luteinizing hormone (LH) in a number of species, including rats, sheep, cattle, and non-human primates. Specifically, endotoxin administration decreases circulating concentration of LH and LH pulses frequency in castrated male sheep. Endotoxin could alter circulating concentrations of LH via actions at the hypothalamus through altered GnRH production and/or release, or endotoxin could alter circulating concentrations of LH at the level of the pituitary via inhibition of LH production and release or inhibition of LH in response to GnRH. The site of endotoxin suppression of circulating concentrations of LH as well as possible mediators of endotoxin suppression of circulating concentrations of LH, including cortiocotropin-releasing hormone, arginine vasopressin, glucocorticoids, inflammatory cytokines, prostaglandins, and opioids, are discussed.     

Growth hormone stimulation of serum insulin concentration in cattle: Nutritional dependency and potential mechanisms

Previous studies on the effect of growth hormone (GH) on serum insulin concentration in cattle had generated seemingly conflicting results, and little was known about the mechanism by which GH affects serum insulin concentration in cattle, if it does. In this study, we determined whether the effect of GH on serum insulin concentration in cattle could be affected by the nutritional levels of the animal and whether GH increased serum insulin concentration in cattle by directly stimulating insulin release or insulin gene expression in the pancreatic islets. Administration of recombinant bovine GH increased serum insulin concentration in nonlactating, nonpregnant beef cows fed a daily concentrate meal in addition to ad libitum hay, but it had no effect in those cows fed hay only. Both GH treatments for 1 and 24 h increased insulin concentrations in cultures of pancreatic islets isolated from growing cattle. Growth hormone treatment for 24 h increased insulin mRNA expression in cultured bovine pancreatic islets. Growth hormone treatment for 16 h increased reporter gene expression directed by a ∼1,500-bp bovine insulin gene promoter in a rat insulin-producing β cell line. Taken together, these results suggest that exogenous GH can increase serum insulin concentration in cattle, but this effect depends on the nutritional levels of fed cattle, and that GH increases serum insulin concentration in cattle by stimulating both insulin release and insulin gene expression in the pancreatic islets.     

Kisspeptin‐10 stimulates the release of luteinizing hormone and testosterone in pre‐ and post‐pubertal male goats

The aims of the present study were to clarify the effect of kisspeptin‐10 (Kp10) on the secretion of luteinizing hormone (LH) and testosterone (T) in pre‐pubertal and post‐pubertal male ruminants. Four male goats (Shiba goats) were given an intravenous (i.v.) injection of Kp10 (5 µg/kg body weight (b.w.)), gonadotoropin‐releasing hormone (GnRH, 1 µg/kg b.w.), or 2 mL of saline as a control at the ages of 3 (pre‐pubertal) and 6 (post‐pubertal) months. A single i.v. injection of Kp10 significantly stimulated the release of LH and T in both groups. The area under the response curve (AUC) of LH for a 60‐min period after the i.v. injection of Kp10 was significantly greater in the pre‐pubertal goats (P < 0.05). The AUC of T for a 120 min period post‐injection did not differ between the two age groups. A single i.v. injection of GnRH also significantly stimulated the release of LH and T in both groups (P < 0.05). The secretory pattern of LH and T in response to GnRH resembled that in response to Kp10. These results show that the LH‐releasing response to Kp10 is greater in pre‐pubertal than post‐pubertal male goats. They also show that Kp10, as well as GnRH, is able to stimulate the release of T in male goats.     

Effect of melatonin injected into the third ventricle on growth hormone secretion in Holstein steers

The effects of melatonin (MEL) injection into the third ventricle (3V) on growth hormone (GH) secretion were investigated in conscious Holstein steers. A stainless steel cannula was stereotaxically implanted in the 3V based on the ventriculogram. In Exp. 1, three doses of MEL (100, 300 or 600 microg) were injected into the 3V through the cannula and the GH concentration after the injection was determined. In Exp. 2, intracerebroventricular (icv) and intravenous (iv) injections of MEL (100 microg) and GH-releasing hormone (GHRH; 0.25 microg/kg body weight), respectively, were performed simultaneously to examine the effect of MEL on GHRH-induced GH release. The icv injection of MEL significantly stimulated GH release at 100 microg. The increase in GH concentrations by 100 microg of MEL was persistent. Intravenous injection of GHRH dramatically increased GH release. The injection of MEL did not alter GHRH-induced GH release. These results suggest that MEL stimulates GH secretion possibly through the hypothalamus in cattle.     

The control of adenohypophysial hormone secretion by amino acids and peptides in swine

Several different amino acids and peptides control secretion of adenohypophysial hormones and this control may be indirect, via the modulation of hypothalamic hormone secretion. Indeed, classical hypothalamic hormones (e.g., gonadotropin-releasing hormone [GnRH], growth hormone-releasing hormone [GHRH], somatostatin, etc.) may be released into the hypothalamo-hypophysial portal vasculature, travel to the adenohypophysis and there stimulate or inhibit secretion of hormones. Alternatively, some amino acids and peptides exert direct stimulatory or inhibitory effects on the adenohypophysis, thereby impacting hormone secretion. In swine, the most extensively studied modulators of adenohypophysial hormone secretion are the excitatory amino acids (ExAA), namely glutamate and aspartate, and the endogenous opioid peptides (EOP). In general, excitatory amino acids stimulate release of luteinizing hormone (LH), follicle-stimulating hormone (FSH), growth hormone (GH), and prolactin (PRL). Secretion of adenohypophysial hormones induced by ExAA is primarily, but perhaps not exclusively, a consequence of action at the central nervous system. By acting primarily at the level of the central nervous system, EOP inhibit LH secretion, stimulate GH release and depending on the animal model studied, exert either stimulatory or inhibitory influences on PRL secretion. However, the EOP also inhibited LH release by direct action on the adenohypophysis. More recently, peptides such as neuropeptide-Y (NPY), orexin-B, ghrelin, galanin, and substance P have been evaluated for possible roles in controlling adenohypophysial hormone secretion in swine. For example, NPY, orexin-B, and ghrelin increased basal GH secretion and modulated the GH response to GHRH, at least in part, by direct action on the adenohypophysis. Secretion of LH was stimulated by orexin-B, galanin, and substance P from porcine pituitary cells in vitro. Because the ExAA and various peptides modulate secretion of adenohypophysial hormones, these compounds may play an important role in regulating swine growth and reproduction.     

Effect of CD14/TLR4 antagonist on GnRH/LH secretion in ewe during central inflammation induced by intracerebroventricular administration of LPS

Background: Immune stress induced by lipopolysaccharide(LPS) influences the gonadotropin-releasing hormone(GnRH)/luteinizing hormone(LH) secretion. Presence of LPS interacting Toll-like receptor(TLR) 4 in the hypothalamus may enable the direct action of LPS on the GnRH/LH secretion. So, the aim of the study was to investigate the influence of intracerebroventricular(icv) injection of TLR4 antagonist on GnRH/LH secretion in anestrous ewes during LPS-induced central inflammation. Animals were divided into three groups icv-treated with: Ringer-Locke solution, LPS and TLR4 antagonist followed by LPS.Results: It was demonstrated that TLR4 antagonist reduced LPS-dependent suppression of GnRH gene expression in the preoptic area and in the medial basal hypothalamus, and suppression of receptor for GnRH gene expression in the anterior pituitary gland. It was also shown that TLR4 antagonist reduced suppression of LH release caused by icv injection of LPS. Central administration of LPS stimulated TLR4 gene expression in the medial basal hypothalamus.Conclusions: It was indicated that blockade of TLR4 prevents the inhibitory effect of centrally acting LPS on the GnRH/LH secretion. This suggests that some negative effects of bacterial infection on the hypothalamic-pituitary-gonadal axis activity at the hypothalamic level may be caused by central action of LPS acting through TLR4.     

小尾寒羊FSHβ和LHβ基因在生殖轴的表达研究

为揭示FSHβ和LHβ基因在小尾寒羊下丘脑-垂体-卵巢轴(HPOA)中的表达规律,深入了解其对小尾寒羊多羔的作用,本研究采用实时荧光定量PCR技术对6只小尾寒羊(FecB++型单、多羔羊各3只)的生殖组织及脑组织中FSHβ和LHβ基因的表达差异进行分析。结果表明:FSHβ和LHβ基因在大脑、小脑、下丘脑、卵巢、子宫、输卵管和垂体7种组织中均有表达,FSHβ主要在小尾寒羊下丘脑和卵巢高表达,LHβ在垂体高表达;FSHβ基因在小尾寒羊多羔群体下丘脑、卵巢、子宫、输卵管、垂体的表达极显著高于单羔群体(P<0.01),LHβ基因在小尾寒羊多羔群体下丘脑、卵巢、子宫、输卵管、垂体、小脑、大脑表达量均极显著高于单羔群体(P<0.01)。研究结果提示,FSHβ和LHβ基因可能参与小尾寒羊多羔性状调控。     

Injection of neuropeptide Y into the third cerebroventricle differentially influences pituitary secretion of luteinizing hormone and growth hormone in ovariectomized cows.

Hypothalamic neurons that control the luteinizing hormone (LH) and growth hormone (GH) axes are localized in regions that also express neuropeptide Y (NPY). Increased hypothalamic expression of NPY due to diet restriction has been associated with suppressed secretion of LH and enhanced secretion of GH in numerous species. However, these physiological relationships have not been described in cattle. Thus, two studies were conducted to characterize these relationships using ovariectomized (Experiment 1) or ovariectomized estrogen-implanted (Experiment 2) cows. In Experiment 1, four well-nourished, ovariectomized cows received third cerebroventricular (TCV) injections of 50 and 500 micrograms of NPY in a split-plot design. Venous blood was collected at 10-min intervals from -4 hr (pre-injection control period) to +4 hr (postinjection treatment period) relative to TCV injection. NPY suppressed (P < or = 0.04) tonic secretion of LH irrespective of dose and tended to stimulate (P < or = 0.10) an increase in tonic secretion of GH. In Experiment 2, six ovariectomized cows that were well nourished and implanted with estradiol received TCV injections of 0, 50, or 500 micrograms of NPY in a replicated 3 x 3 Latin Square. Both doses of NPY suppressed (P < 0.06) mean concentration of LH relative to the 0-microgram dose. The 50-microgram dose of NPY tended (P < 0.10) to increase the amplitude of GH pulses. In conclusion, TCV injection of NPY suppressed pituitary secretion of LH and simultaneously tended to increase pituitary secretion of GH.     

Endogenous opiates and the hypothalamic-pituitary-gonadal axis in male sheep

The effects of morphine and the opiate receptor antagonist, naloxone, on the secretory pattern of luteinizing hormone (LH) were assessed in male sheep. Morphine infusion (250 mg/hr) abruptly stopped LH pulsatile secretion in castrates (wethers) and decreased mean serum LH concentrations by nearly 70 percent. Response of the pituitary to exogenous LH releasing hormone was not affected by morphine suggesting that the effects of morphine on LH secretion were mediated through the hypothalamus. Estradiol-implanted wethers, characterized by a nonpulsatile LH secretory pattern, responded to intravenous injection of naloxone (20, 50 and 200 mg Lv.) with an immediate release (pulse) of L.H. Similarly, LH release was significantly increased following naloxone infusion (200 mg/hr for four hours) in intact rams and wethers implanted with testosterone or estradiol. In contrast, naloxone infusion altered the pattern of LH secretion in wethers but without affecting mean serum LH concentrations. These results support the notion that LH secretion in male-sheep is tonically regulated by endogenous opiates and further suggests that opioid modulation of the hypothalamic-pituitary-LH axis in sheep involves an interaction with the steroid negative feedback system.     

Comparison of the ability of the three endogenous GnRHs to stimulate release of follicle-stimulating hormone and luteinizing hormone in chickens

It is well established that GnRH can stimulate the release of LH and FSH in mammals. Two GnRHs have been found in the chicken hypothalamus, cGnRH-I and -II. There is controversy as to whether either peptide can stimulate release of FSH in birds. The present studies compared the ability of cGnRH-I and -II to stimulate the release of FSH and LH in chickens. Lamprey (l) GnRH-III may be a specific-releasing factor for FSH, as it selectively stimulates FSH release in rodents and cattle, and has been detected in the hypothalamus of rodents, sparrows and chickens. Therefore, the ability of lGnRH-III to stimulate LH and FSH release was also examined. In our first experiment, the effects of cGnRH-I and -II were studied using 17-week prepubertal females. Intravenous injection of cGnRH-II at 1 and 10 microg/kg BW significantly increased LH secretion more than did cGnRH-I. Neither peptide significantly increased plasma FSH levels. In our second study, we administered cGnRH-I, -II or lGnRH-III to mature males maintained on a short photoperiod. cGnRH-II was again more potent than cGnRH-I in stimulating LH release, while lGnRH-III produced a modest LH rise. No GnRH peptide provided specific or potent stimulus to FSH secretion, although the high dose of cGnRH-II modestly enhanced FSH levels in the adult male (P < 0.05). Our results are not consistent with the view that lGnRH-III is a specific FSH-releasing hormone across multiple classes of vertebrates. We conclude that the mechanism by which independent release of FSH occurs in chickens remains unresolved.     

Adrenergic regulation of growt hormone secretion in the ewe

This study examined the role of the adrenergic system in the regulation of growth hormone (GH) secretion in sheep. Intravenous infusion of noradrenaline (0.5μg/kg per min for 2 hr) totally suppressed plasma GH concentrations. Concomitant treatment of animals with the β-adrenergic antagonist propranolol completely blocked the noradrenaline-induced suppression of GH. In contrast, intravenous injection of the centrally acting α₂-agonist clonidine (2μg/kg) elicited a release of GH. To further investigate the central adrenergic regulation of GH secretion 10 μg of noradrenaline or adrenaline was microinjected (1μl) directly into the preoptic area of the hypothalamus of ovariectomized ewes. When the time of injection coincided with a GH trough period, both noradrenaline and adrenaline caused an increase in plasma GH concentrations, whereas if the injection coincided with an endogenous pulse of GH no additional GH response was obtained. In conclusion, these results provide evidence for the involvement of the adrenergic system in the regulation of GH secretion in sheep. Centrally, adrenergic pathways exert a stimulatory effect on GH release via an α₂-adrenergic system, whereas peripherally adrenergic pathways exert an inhibitory effect via β-adrenergic mediated mechanisms. Furthermore, adrenergic stimulation of the preoptic area may inhibit somatostatin activity and directly facilitate a GH pulse. Alternatively, adrenergic innervation of the preoptic area may influence neurons (somatostatin or other) that project to the arcuate nucleus and stimulate the release of GH-releasing factor.     

Effects of dietary energy on control of luteinizing hormone secretion in cattle and sheep.

Prolonged restriction of dietary energy delays onset of puberty, disrupts cyclicity in sexually mature animals, and lengthens the postpartum anestrous period in domestic ruminants. One important mechanism by which energy restriction impairs reproductive activity seems to be suppression of the increase in LH pulse frequency that is necessary for growth of ovarian follicles to the preovulatory stage. Under-nutrition apparently inhibits pulsatile secretion of LH by reducing LHRH secretion by the hypothalamus. The ability of an animal to sustain a high-frequency mode of pulsatile LH release is related to its metabolic status. Mechanisms linking metabolic status to LHRH secretion have not been fully characterized. Changes in body fat have been associated with changes in reproductive activity, but it is unlikely that body fat per se regulates LHRH secretion. It is possible that pulsatile LHRH release is regulated by specific metabolites and(or) metabolic hormones that reflect nutritional status. Alternatively, availability of oxidizable metabolic fuels, such as glucose and nonesterified fatty acids, may influence activity of neurons that control LHRH release. Our understanding of how the central nervous system transduces information about nutritional status into neuroendocrine signals that control reproduction in cattle and sheep is limited by a lack of information concerning the nature of neurons controlling LHRH release in these species.     

Cry1基因在雄性绵羊生殖轴系的表达与定位

隐花色素1(Cryptochrome 1,Cry1)基因作为生物钟调控环路的负调控因子,对生物节律的稳定发挥着重要作用,因此研究Cry1基因在哺乳动物生殖轴系的表达对揭示哺乳动物季节性繁殖的调控机理有着重要意义。本实验应用qPCR和免疫组织化学等方法研究了Cry1基因在雄性绵羊生殖轴系(松果体、下丘脑、垂体、睾丸和附睾)的分布和表达情况,并通过生物信息学分析对Cry1蛋白的结构做了预测。结果显示:在绵羊生殖轴系各组织中均有Cry1表达,其中在下丘脑中的表达最高,睾丸次之,各组数据间差异显著。免疫组化结果显示Cry1蛋白在松果体和下丘脑中成弥散性表达,胞膜、胞质及胞核均有阳性表达。在垂体上Cry1蛋白主要位于毛细血管的管壁细胞和血管周围的腺细胞上。Cry1蛋白还在睾丸组织的基膜和间质细胞,以及附睾管腔上皮细胞、管周肌样细胞和腔面精子上呈阳性表达。生物信息学分析发现Cry1蛋白无信号肽信息和跨膜结构,主要位于细胞质和细胞核中,与牛、鹿的亲缘关系最近。Cry1基因参与了绵羊的繁殖,为生物钟调控哺乳动物的季节性繁殖提供了一定的研究基础。