Ontogeny of the hypothalamo-pituitary-adrenocortical axis in the chicken embryo: a review

The embryo of the domestic fowl (Gallus domesticus) tenders one distinctive advantage over general mammalian models for investigating the development of the hypothalamo–pituitary–adrenocortical (HPA) axis. This is the relative simplicity with which the embryonic endocrine environment can be influenced without confounding maternal influences. The ease of direct manipulation of the embryonic endocrine system has facilitated analysis of the development and function of the HPA axis in the chick embryo. As the chick embryo develops, functional activation of the adrenal gland is regulated at three different levels: the adrenal gland itself, the anterior pituitary, and the hypothalamus. The adrenal gland appears capable of independent secretion of glucocorticoids from day 8 until shortly after day 14 of embryonic development, at which point the pituitary influences adrenocortical activity. Around the same age, the hypothalamic level of control also begins. The information covered in this review will describe the major steps in the development of the HPA axis in the chicken embryo and show that the chicken has an emblematic HPA neuroendocrine axis.     

Pituitary receptors for GnRH and estradiol, and pituitary content of gonadotropins in beef cows. II. Changes during the postpartum period

Pregnant beef heifers (n = 24) were assigned randomly to four groups and slaughtered at day 1, 15, 30 or 45 postpartum. The day prior to slaughter blood samples were taken from each cow every 15 min for 8 hr. The anterior pituitary gland, preoptic area (POA) and medial basal hypothalamus (HYP) were collected from each cow. Contents of gonadotropin-releasing hormone (GnRH) in extracts of POA and HYP, and luteinizing hormone (LH) and follicle-stimulating hormone (FSH) in extracts of anterior pituitary were quantified by radioimmunoassay. In the anterior pituitary gland, membrane receptors for GnRH were quantified by a standard curve technique and cytosolic receptors for estradiol were quantified by saturation analysis. Concentrations of LH, FSH and prolactin in serum were quantified by radioimmunoassay. Only one cow of eight had a pulse of LH during the 8 hr bleeding period on day 1 postpartum. This increased to 8 pulses in 6 cows on day 30 postpartum. Contents of GnRH in POA (15.0 +/- 3.2 ng) and HYP (14.0 +/- 2.0 ng) did not change significantly during the postpartum period. Pituitary content of LH was low following parturition (.2 +/- .1 mg/pituitary) and increased significantly through day 30 postpartum (1.2 +/- .1 mg/pituitary). Pituitary content of FSH did not change over the postpartum period. Receptors for both GnRH (.9 +/- .2 pmoles/pituitary) and estradiol (5.0 +/- .9/moles/pituitary) were elevated on day 15 postpartum, possibly increasing the sensitivity of the anterior pituitary gland to these hormones and leading to an increased rate of synthesis of LH that restored pituitary content to normal by day 30 postpartum.     

Pituitary receptors for GnRH and estradiol, and pituitary content of gonadotropins in beef cows. I. Changes during the estrous cycle

To further characterize the endocrinological changes in the hypothalamo-hypophyseal axis thoughout the bovine estrous cycle, cycling beef heifers (n = 24) were randomly assigned to six groups. These heifers were slaughtered 6, 12, 18, 19, 20 or 21 days following their previous estrus (day 0). Anterior pituitaries and hypothalami were collected. Hypothalami were divided into the preoptic area and medial basal hypothalamus, and content of gonadotropin-releasing hormone (GnRH) was quantified by radioimmunoassay. Contents of luteinizing hormone (LH) and follicle stimulating hormone (FSH) in the anterior pituitary gland were quantified by radioimmunoassay. Membrane receptors for GnRH were quantified by a standard curve technique and receptors for estradiol in anterior pituitary cytosol were quantified by saturation analysis. There was no significant change in content of GnRH in the hypothalamus or content of FSH in the anterior pituitary on any of the days examined; however, content of GnRH in the preoptic area was lower (P less than .1) on day 19 postestrus. Cytosolic receptors for estradiol increased (P less than .05) on day 18 post-estrus and returned to baseline by day 19. Content of LH and the number of receptors for GnRH in the anterior pituitary gland decreased (P less than .01) on day 19 postestrus, and the number of receptors for GnRH remained low through day 21 postestrus. The reduction in anterior pituitary content of LH was transient indicating that synthesis of LH restores pituitary content to preovulatory levels before the number of receptors for GnRH returns to normal.     

The immediate effects of weaning stress on the hypothalamus‐pituitary‐adrenal alteration of newly weaned piglets

This study was conducted to examine the effects of weaning stress on gene expression of specific markers in hypothalamus‐pituitary‐adrenal (HPA) axis and neuronal activity in the newly weaned piglets. Twelve 28‐days‐old, newly weaned crossbred (Landrace × Yorkshire × Duroc) male piglets from 6 l (2 piglets/l) were randomly categorized into two groups: (a) weaning stress: piglets were separated from their dams, relocated and mixed with the unacquainted domestic piglets for 2 hr (stress, n = 6); (b) no‐stress: piglets stayed with their dams in the farrowing house (NS; n = 6). After weaning stress, all piglets were electrically euthanized and the blood samples/HPA tissues were collected for subsequent analysis, including plasma cortisol and mRNA expression of c‐fos, c‐jun, corticotropin‐releasing hormone (CRH), CRH receptor 1 (CRHR‐1) and adrenocorticotropin hormone receptor (MC2R). Results: Weaning stress significantly (p < 0.05) increased the plasma cortisol level and suppressed the expression of c‐fos and CRH in hypothalamus. In addition, weaning stress enhanced the mRNA abundance of c‐jun and CRHR‐1 in the pituitary gland. No significant differences in the gene expression of MC2R and CRHR‐1 were observed in the adrenal gland between treatment groups. Taken together, HPA involved in weaning stress and CRHR‐1 and c‐jun could be potential markers to evaluate the activation of HPA axis.     

脂多糖刺激对断奶仔猪下丘脑-垂体-肾上腺轴PPARγ表达的影响

研究脂多糖(LPS)对断奶仔猪下丘脑-垂体-肾上腺(HPA)轴过氧化物酶体增殖物活化受体γ(PPARγ)表达的影响。对照组注射生理盐水,试验组注射LPS。注射后1.5 h和3 h采血,3 h采血后屠宰。结果表明:LPS刺激后1.5 h,中性粒细胞含量及其比例显著下降(P<0.05);LPS刺激后3 h,白细胞、淋巴细胞、中性粒细胞含量显著下降(P<0.05)。LPS刺激后1.5 h,血浆肿瘤坏死因子(TNF)-α、皮质醇和促肾上腺皮质激素释放素激素(CRH)含量显著上升(P<0.05);LPS刺激后3 h,血浆TNF-α、皮质醇和促肾上腺皮质激素(ACTH)含量显著上升(P<0.05);LPS刺激导致下丘脑、腺垂体、肾上腺皮质和髓质中PPARγ阳性细胞百分率显著升高(P<0.05)。这表明LPS导致免疫应激,激活HPA轴,诱导HPA轴PPARγ的表达。     

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.     

Failure of neuropeptide y to modulate the release of LH and prolactin by cultured bovine pituitary cells

Possible direct effects of neuropeptide Y (NPY) on dispersed and cultured cells of the anterior lobe (AL) of the bovine pituitary were investigated. AL tissue from steers was enzymatically dissociated into individual cells, preincubated for 18 hr and then incubated in suspension cultures for 2 hr or 24 hr with either NPY, gonadotropin-releasing hormone (GnRH) or both. Release of luteinizing hormone (LH) and prolactin (PRL) into medium was quantified by radioimmunoassay and expressed as hormone released per 100,000 cells. Basal release of LH averaged 38 and 86 ng for 2 hr and 24 hr respectively while that of PRL averaged 118 and 438 ng for the same incubation periods. Addition of NPY did not alter (P>.05) basal release of LH or PRL for either duration of incubation. Also, NPY did not affect (P>.05) release of LH in response to GnRH. In summary, this study indicated that NPY, at in vitro dosages of .01 to 100nM, does not modulate the release of LH or PRL at the pituitary level in castrate cattle.     

Effects of season and sex on in vitro aromatase and 17β-oxidoreductase activities in the brain and anterior pituitary gland of gonadectomized sheep

To improve understanding of seasonal control of reproduction, we studied effects of season and sex on in vitro conversion of androstenedione to estrone and testosterone within the brain and anterior pituitary gland of recently gonadectomized sheep. For both sexes, aromatase activity was relatively high in the amygdala and hypothalamus plus preoptic area (HPOA), but aromatase was not detected in the parietal cortex or the anterior pituitary gland. Aromatase activity of the amygdala was not affected by sex or season. For castrated rams, aromatase activity in the HPOA in May was <25% of that in December (P<.05). For ovariectomized ewes, however, aromatase activity in the HPOA was similar in May and December (P>.05). All tissues contained 17β-oxidoreductase. For the amygdala and HPOA, conversion of androstenedione to testosterone was greater (P<.05) in the male than the female, regardless of season. In rams, 17β-oxidoreductase activity was lower in the HPOA and higher in the anterior pituitary gland in May than in December. In the anterior pituitary of ewes, however, 17β-oxidoreductase activity was higher in December. Thus, conversion of androstenedione to testosterone and estrogens within the brain and anterior pituitary gland could help to control the secretion of luteinizing hormone. We postulate that in rams a seasonal reduction of aromatase activity in the HPOA and increase in 17β-oxidoreductase in the anterior pituitary gland during May may be involved with onset of the seasonal increase in LH secretion.     

Development, validation, and utilization of a novel antibody specific to the type III chicken gonadotropin-releasing hormone receptor

Two gonadotropin-releasing hormone receptors (GnRH-Rs) have been characterized in chickens to date: cGnRH-R-I and cGnRH-R-III, with cGnRH-R-III being the predominant pituitary form. The purpose of the present study was to first validate a novel antibody for the specific detection of cGnRH-R-III and second, using this antibody, detect changes in cGnRH-R-III protein levels in the pituitary gland of male and female chickens during a reproductive cycle. The localization of cGnRH-R-III within the anterior pituitary gland was also determined. Western blotting of pituitary extracts and transiently transfected COS-7 cell lysates revealed that our antibody is highly specific to cGnRH-R-III protein. Similarly, when used in immunocytochemistry, this antibody specifically detects cells expressing cGnRH-R-III and not cGnRH-R-I. Western blot analyses of chicken pituitary gland homogenates show that cGnRH-R-III protein levels are significantly greater in sexually mature birds than in immature birds or birds at the end of a reproductive cycle (P < 0.0001). A similar pattern was observed for both males and females. Additionally, the antibody was able to detect cGnRH-R-III in cells along the periphery of the cephalic and caudal lobes of the anterior pituitary where the cells containing the gonadotropins are located. In summary, we successfully validated a novel antibody to cGnRH-R-III and showed levels of cGnRH-R-III protein in the pituitary fluctuate with respect to the reproductive status in both male and female chickens.     

Association study of molecular polymorphisms in candidate genes related to stress responses with production and meat quality traits in pigs

The hypothalamic-pituitary-adrenal (HPA) axis exerts a large range of effects on metabolism, the immune system, inflammatory processes, and brain functions. Together with the sympathetic nervous system, it is also the most important stress-responsive neuroendocrine system. Both systems influence production traits, carcass composition, and meat quality. The HPA axis may be a critical target for genetic selection of more robust animals. Indeed, numerous studies in various species have demonstrated the importance of genetic factors in shaping the individual HPA axis phenotype, and genetic polymorphism can be found at each level of the axis, including hormone production by the adrenal cortices under stimulation by adrenocorticotropic hormone (ACTH), hormone bioavailability, or receptor and postreceptor mechanisms. The aim of the present experiment was to extend these findings to the brain neurochemical systems involved in stress responses. To this end, a number of candidate genes were sequenced for molecular polymorphisms and their association was studied with stress neuroendocrine and production traits in a genetically diverse population consisting of 100 female pigs from an advanced intercross (F10-F12) between 2 highly divergent breeds, Large White (LW) and Meishan (MS). The LW breed has a high production potential for lean meat and a low HPA axis activity, and the MS breed has low growth rate, fat carcasses—but large litters of highly viable piglets—and a high HPA axis activity. Candidate genes were chosen in the catecholaminergic and serotonergic pathways, in the pituitary control of cortisol production, among genes previously demonstrated to be differentially expressed in ACTH-stimulated adrenal glands from LW and MS pigs, and in cortisol receptors. Sixty new polymorphisms were found. The association study with carcass and meat quality traits and with endocrine traits showed a number of significant results, such as monoamine oxidase (MAOA) polymorphisms with growth rate (P = 0.01); lean content and intramuscular fat (P < 0.01), which are the most important traits for carcass value; dopamine receptor D3 (DRD3) with carcass composition (P < 0.05); and vasopressin receptor 1B (AVPR1B) with meat quality traits (P ≤ 0.05). The effect of these polymorphisms on neuroendocrine parameters (eg DRD3 and HPA axis or AVPR1B and catecholamines) indicates information regarding their biological mechanism of action.     

发情期和乏情期哈萨克羊垂体差异表达基因筛选及功能预测

垂体是哺乳动物重要的内分泌器官之一,主要通过调控激素合成来调节动物发情。为了研究绵羊垂体系统在发情状态和乏情状态之间的基因表达差异,本研究使用RNA-seq技术分析了发情期和乏情期哈萨克羊垂体前叶的转录组数据。通过比较转录组数据发现:在发情期和乏情期的哈萨克羊垂体中共筛选出了3 211个差异表达的基因,其中包含298个上调的和2 913个下调的基因。利用GO和KEGG富集分析这些差异表达的基因发现:它们被显著地富集在卵母细胞减数分裂和孕酮介导的卵母细胞成熟等调控发情的信号通路上,提示这些基因在调控哈萨克羊发情状态中发挥着不可或缺的作用。本研究丰富了绵羊的转录组资源,为今后深入研究绵羊季节发情机制、提高绵羊繁殖率打下了良好的基础。     

Hypothalamic-pituitary-adrenal axis responsiveness to insulin-induced hypoglycaemia is modified by trypanosome infection in Boran (Bos indicus) cattle.

Ten Boran (Bos indicus) cattle were used to study the stress responsiveness of the hypothalamic-pituitary-adrenal (HPA) axis during trypanosome infection. Five cattle were infected with Trypanosoma congolense IL 1180 by tsetse challenge and five cattle served as controls. All infected animals developed acute trypanosomiasis. Insulin-induced hypoglycaemia (50 per cent of pre-insulin glucose concentration) was used as a stress factor. Acute hypoglycaemia was observed in three infected and three control animals after insulin challenge. Two animals from each group either did not respond or responded slowly. Hypoglycaemia in infected animals completely failed to induce an HPA axis response, while in control animals an HPA axis response was indicated by a significant increase in plasma adrenocorticotrophic hormone (ACTH) and cortisol concentrations (P less than 0.01). The results show that trypanosomiasis in Boran cattle can cause a decrease in the stress responsiveness of the HPA axis as indicated by a blunted ACTH/cortisol response to insulin-induced hypoglycaemia.     

Changes in the concentration of hypothalamic and hypophyseal receptors for estradiol in pregnant and postpartum ewes

The sensitivity of the hypothalamic-pituitary axis to estradiol was evaluated by monitoring changes in the concentration of receptors for estradiol during gestation and the postpartum period of the ewe. Ewes that had been bred during anestrus so that lambing would occur during the breeding season were killed on d 50 and 140 of gestation or on d 2, 13, 22 and 35 postpartum. Total cytosolic receptors for estradiol were quantified in the anterior and posterior hypothalamus, median eminence, anterior pituitary and cerebral cortex. Concentration and total number of receptors for estradiol in the anterior and posterior hypothalamus and anterior pituitary gland during gestation through d 13 postpartum were similar but increased (P less than .05) by d 22 and 35 postpartum. Changes in the concentration of receptors for estradiol were tissue-specific because stage of reproduction did not influence the number of receptors for estradiol in the median eminence or cerebral cortex. Increases in the concentration of receptors for estradiol in hypothalamic and pituitary tissues on d 22 postpartum immediately preceded the onset of normal ovarian cyclicity in ewes. From these data we conclude that resumption of reproductive cycles in postpartum ewes occurs about the time concentrations of receptors for estradiol in the anterior pituitary gland and hypothalamus increase.     

母体妊娠期应激对后代出生前后下丘脑-垂体-肾上腺轴的影响及其机制

动物的生理健康与其正常下丘脑-垂体-肾上腺(hypothalamus-pituitary-adrenocortical,HPA)轴活性是密不可分的.母体妊娠期应激对后代出生前后HPA轴功能及生长发育等均有显著的程序化影响,尤其是对胎儿期HPA轴的影响更大.这种影响主要是通过改变后代上游海马体、下丘脑和垂体中的糖皮质激素受体和盐皮质激素受体表达实现的.下游器官肾上腺皮质作为HPA轴的重要组成部分,也可能是HPA轴程序化的靶器官,有待深入研究.     

Feeding-induced increases in insulin do not suppress secretion of growth hormone

Secretion of growth hormone (GH) is reduced for several hours after feeding when access to feed is restricted to a 2-hr period each day. We hypothesized that increased secretion of insulin after feeding inhibits release of GH from the anterior pituitary gland. Our objectives were to determine whether: 1) alloxan prevents concentrations of insulin from increasing after feeding steers; 2) concentrations of GH remain high after feeding alloxan-treated steers; and 3) GH-releasing hormone (GHRH) stimulates greater release of GH in alloxan-treated, than in control, steers after feeding. Steers were injected iv with either saline (control) or with alloxan (110 mg/kg) (n = 4 per group). Concentrations of insulin were not different (P = 0.61) between control and alloxan-treated steers before feeding (87.5 +/- 33.6 pmol/l). However, alloxan prevented insulin from increasing (P < 0.001) after feeding (131.8 pmol/1) compared with control steers (442.0 pmol/l) (pooled SEM = 47.5). Overall, GH was higher (P < 0.05) in alloxan-treated (6.4 ng/ml) than in control steers (3.7 ng/ml) (pooled SEM = 0.7), but GH decreased (P < 0.001) after feeding in both groups. Iv injection of GHRH stimulated release of GH 1 hr before, but not when injected 1 hr after feeding (P < 0.001). In addition, net areas under the GH curve were not significantly different between control and alloxan-treated groups. We conclude that increased concentrations of insulin after feeding do not mediate feeding-induced suppression of GH secretion in steers.     

Feeding-induced increases in insulin do not suppress secretion of growth hormone

Secretion of growth hormone (GH) is reduced for several hours after feeding when access to feed is restricted to a 2-hr period each day. We hypothesized that increased secretion of insulin after feeding inhibits release of GH from the anterior pituitary gland. Our objectives were to determine whether: 1) alloxan prevents concentrations of insulin from increasing after feeding steers; 2) concentrations of GH remain high after feeding alloxan-treated steers; and 3) GH-releasing hormone (GHRH) stimulates greater release of GH in alloxan-treated, than in control, steers after feeding. Steers were injected iv with either saline (control) or with alloxan (110 mg/kg) (n = 4 per group). Concentrations of insulin were not different (P = 0.61) between control and alloxan-treated steers before feeding (87.5 +/- 33.6 pmol/l). However, alloxan prevented insulin from increasing (P < 0.001) after feeding (131.8 pmol/1) compared with control steers (442.0 pmol/l) (pooled SEM = 47.5). Overall, GH was higher (P < 0.05) in alloxan-treated (6.4 ng/ml) than in control steers (3.7 ng/ml) (pooled SEM = 0.7), but GH decreased (P < 0.001) after feeding in both groups. Iv injection of GHRH stimulated release of GH 1 hr before, but not when injected 1 hr after feeding (P < 0.001). In addition, net areas under the GH curve were not significantly different between control and alloxan-treated groups. We conclude that increased concentrations of insulin after feeding do not mediate feeding-induced suppression of GH secretion in steers.     

NDV对雏鸵鸟HPA轴的影响

以45日龄雏鸵鸟为试验动物,分正常组和攻毒组,利用RIA技术和TUNEL技术,探讨新城疫病毒（NDV）对雏鸵鸟HPA轴的影响。结果表明,雏鸵鸟感染NDV,HPA轴内细胞凋亡数量在病毒感染期间明显高于对照组,提示NDV能诱导雏鸵鸟HPA轴系统发生细胞凋亡;在病毒接种后1d,肾上腺内可检测到大量凋亡细胞,接种5d后凋亡数量显著增加（P〈0.05）,之后凋亡呈下降趋势,肾上腺内细胞凋亡的动态变化反映了雏鸵鸟HPA轴对ND病变的适应和调节。雏鸵鸟感染NDV,血清ACTH水平于NDV接种后1d开始上升,至5d达到峰值（P〈0.01）,之后有所下降,渐趋于正常;血清Cor水平于病毒接种后1d开始下降,至7d有所回升,9d渐趋于正常水平,表明NDV接种后雏鸵鸟血清ACTH和Cor水平的变化与其HPA轴功能的损伤及恢复密切相关。     

Pituitary weight and gonadotrophin concentration, body weight and ovarian weight in growing and mature female turkeys exposed to different light regiments

Anterior pituitary weights and gonado‐trophin concentration, body weight changes and ovarian weights were followed every two weeks in three lighting treatment periods in spring hatched turkeys. In the growing period (12 to 20 weeks of age), all of the characters changed with the increasing age of the birds while no changes were associated with the lighting treatments (either 14 hr of light daily or natural daylight). During the conditioning period (20 to 28 weeks) anterior pituitary weight, body weight change and ovarian weight all increased with increasing age. No changes were found in anterior pituitary gonadotrophin concentration. Body weight change and ovarian weight were also greater in birds maintained on range as compared with birds treated with 6 or 14 hr of light daily. In the production period (28 to 40 weeks), the birds were divided into laying and non‐laying groups. In the laying group, no differences were found in the three characters studied (ovarian weight not included) within the ages studied. In the non‐laying group, the birds on range had greater body weight increases within the 34th to 38th week than the birds on the other lighting treatments. More of the birds given 6 hr of light during the conditioning period were classified as layers in the production period than birds treated with either 14 hr or natural light (14.3 decreasing to 11.5 hr) during the conditioning period. Once a bird had started laying, a decided drop in anterior pituitary gonadotrophin occurred.