家畜免疫去势研究进展

传统的手术去势技术正受到动物福利和规模化标准化畜牧生产需求的挑战,作为手术去势的优势替代方法,免疫去势能够很好地阻滞睾丸发育,控制性行为,避免手术带来的痛苦。免疫去势是采用免疫学方法破坏下丘脑-垂体-性腺轴的激素平衡,通过降低促性腺激素释放激素(GnRH)、黄体生成素(LH)和卵泡刺激素(FSH)的水平,进而使性腺激素水平降低,最终抑制性腺功能,达到去势的目的。免疫去势分为激素免疫去势和基因免疫去势,其靶标经历了下丘脑-垂体-性腺轴自下而上的筛选,依次为睾酮、LH、FSH、GnRH、吻素-1(KISS-1),其中位于下丘脑-垂体-性腺轴上游的GnRH激素、GnRH和KISS-1基因3种靶标去势效果较好。免疫去势效果及机制的相关研究表明,免疫去势具有改善生产性能、安全和可逆的特点,但其可逆性机制还不清楚。去势导致肾上腺发挥代偿作用,但目前免疫去势的肾上腺代偿研究鲜见报道。文章首先介绍了几种靶标的免疫去势机制,其次对免疫去势的特点和应用进行了讨论,旨在为推动免疫去势技术在动物生产繁殖领域的应用提供理论依据。     

禽类下丘脑-垂体-性腺轴的内分泌调节

动物生殖系统的发育和功能维持受到下丘脑-垂体-性腺（HPG）轴的调控。下丘脑、垂体、性腺在中枢神经的调控下形成一个封闭的自动反馈系统，三者相互协调、相互制约使动物的生殖内分泌系统保持相对稳定。下丘脑接受经中枢神经系统分析与整合后的各种信息，以间歇性脉冲形式分泌促性腺激素释放激素（GnRH），刺激垂体前叶分泌促性腺激素（GTH），即卵泡刺激素（FSH）和黄体生成素（LH），然后促进睾丸或卵巢的发育并分泌睾酮或雌二醇。性腺、垂体、下丘脑释放的调控因子又可以作用于上级中枢或其自身，形成长轴、短轴和超短轴反馈调节通路。     

羊促性腺激素释放激素主动免疫去势的研究进展

促性腺激素释放激素(GnRH)是由下丘脑神经内分泌小细胞产生的十肽激素,主要通过下丘脑-垂体-性腺轴(HPG)参与调控动物的生殖活动,也可直接作用于性腺或其他器官发挥重要功能。GnRH免疫去势作为当前一种动物友好的去势方法在生产中应用和推广。本文综述了GnRH的分子结构和生物学功能、GnRH主动免疫的应用、GnRH免疫在改善公羊生产性能和繁殖性能上的研究进展,为进一步研究GnRH主动免疫在公羊繁殖中的作用提供一定理论支撑,以提高羊养殖业的经济效益和社会效益。     

哺乳动物下丘脑-垂体-卵巢轴的研究进展

哺乳动物的下丘脑、垂体和卵巢分泌的激素在功能上相互作用,构成一个完整的神经内分泌生殖调节体系,即下丘脑垂体卵巢轴,它在生殖活动中起着主要的调节作用。下丘脑中分布的GnRH神经元可以分泌GnRH,GnRH调节垂体中促性腺激素细胞分泌促性腺激素FSH和LH,促性腺激素作用于卵巢受体,引起雌激素和孕酮分泌并影响生殖活动。从组织学角度上研究,下丘脑垂体卵巢轴中的结构,如GnRH神经元、促性腺激素细胞、卵泡随周期性变化而呈现出不同的形态结构和分泌特点。因此,对以上各种细胞的研究是探讨其所分泌激素的基础,而下丘脑垂体卵巢轴中的各种激素的研究则是了解和控制动物繁殖机能的关键。     

GnRH主动免疫对SD雄鼠下丘脑GnRH合成及性腺反馈系统的影响

探讨GnRH主动免疫对SD雄鼠下丘脑GnRH合成及性腺反馈系统的影响。36只SD雄鼠随机分为免疫组、手术去势组及对照组,免疫组于12周龄时初免,8周后加免。放免法测定血清抗体滴度、激素浓度及下丘脑正中隆起GnRH含量,实时荧光定量PCR分析下丘脑生殖相关基因mRNA的变化。结果显示,GnRH主动免疫后12只免疫鼠中11只血清GnRH抗体滴度显著升高,血清LH、FSH及T浓度显著降低(P<0.05),睾丸严重萎缩(免疫去势鼠)。与对照鼠相比,免疫去势显著降低下丘脑GnRH含量(P<0.01),且显著下调下丘脑雌激素α受体(ERα)、雄激素受体(AR)、Kiss-1、GPR54及GnRH的mRNA表达水平(P<0.05)。手术去势鼠除血清LH、FSH浓度及下丘脑ERα的mRAN表达水平显著高于对照鼠外(P<0.05),其余指标均与免疫去势鼠类似。结果首次表明,GnRH主动免疫抑制性腺负反馈调节作用及降低了下丘脑GnRH的合成。     

GnRH的生物学作用及其类似物在马属动物中的应用研究进展

繁殖率低是制约马属动物产业发展的瓶颈,繁殖调控技术不完善也是导致该产业发展缓慢的主要因素。促性腺激素释放激素（gonadotropin-releasing hormone,GnRH）是下丘脑分泌产生的神经激素,在下丘脑-垂体-性腺轴通路中起关键作用。GnRH可以有效促进马属动物卵泡发育及排卵。GnRH是具有十肽结构的化合物,通过改变十肽结构中不同位置的氨基酸合成不同的GnRH类似物。GnRH及其类似物可通过刺激卵泡刺激素（follicle-stimulating hormone,FSH）和黄体生成素（luteinizing hormone,LH）的分泌、抑制雌激素受体的合成影响动物繁殖性能。GnRH及其类似物已被证明可提高马属动物的繁殖性能。对GnRH和促性腺激素释放激素受体（gonadotropin-releasing hormone receptor,GnRHR）的结构特点、生物学功能以及GnRH类似物在马属动物繁殖方面的应用研究进展进行综述,以期为探究GnRH生物学作用的分子机制以及GnRH类似物在马属动物繁殖调控中的科学应用提供参考。     

GnRH主动免疫雌性大鼠调控机制研究

旨在探讨GnRH主动免疫雌性SD大鼠的调控机制。24只雌性SD大鼠随机分为免疫组、卵巢切除组和空白对照组(n=8),免疫组于12周龄注射疫苗,4周后加免;卵巢切除组11周龄外科手术摘除卵巢;空白对照组不作任何处理。使用放射免疫分析法测定血清激素E2、P4、FSH和LH含量,荧光定量PCR分析各基因mRNA表达情况。结果显示,主动免疫显著降低血清E2、P4、FSH和LH含量,至56周处死时,E2和LH含量有微弱上升,但4种激素含量均显著低于空白对照组(P0.05)。与空白对照组相比,主动免疫显著下调下丘脑GPR54、KiSS-1、GnRH和ER-αmRNA,垂体GnRH-R、FSH-β和LH-βmRNA以及卵巢FSHR、LHR和ER-αmRNA的表达水平(P0.05)。结果表明,GnRH主动免疫能显著抑制雌性SD大鼠性腺发育,降低血清性激素含量,通过下丘脑ER-αmRNA-GPR54/KiSS-1通路保持动物较长时间去势;但GnRH主动免疫造成的动物去势存在恢复的可能。     

促性腺激素释放激素及其类似物的功能与应用

促性腺激素释放激素（gonadotropin releasing hormone，GnRH）是主要由下丘脑合成的直链十肽激素，以脉冲形式分泌进入丘脑下部垂体门脉系统到达垂体前叶，或进入神经分泌轴突末端，调控垂体促卵泡激素（follicle-stimulating hormone，FSH）、促黄体素（1uteinizing hormone，LH）的合成与分泌。它是神经、免疫和内分泌调节系统相互联系的重要信号分子，与性腺和胎盘及生殖功能密切相关。     

影响母猪繁殖性能的几种主要因素

1激素 母猪在繁殖过程中,主要受促卵泡素（FSH）和促黄体素（LH）的激素作用。FSH和LH的分泌受下丘脑--垂体轴的调控,同时对下丘脑--垂体轴产生负反馈作用。各种因素对母猪繁殖过程的作用较为复杂,主要通过影响两个途径而发生作用：通过下丘脑--垂体--性腺轴作用影响LH和FSH分泌,从而对卵泡生长发育产生影响;直接作用于卵巢对卵泡细胞的增殖、生长和分化起作用。     

KISS基因的研究进展

KISS-1基因与GPR54基因一起组成KISS-1/GPR54系统,参与动物下丘脑-垂体-性腺轴功能的调节；KISS-2基因也参与动物性腺轴调控,而且作用比KISS-1基因强.研究经济动物KISS基因与繁殖性状的相关有非常重大的意义.本文主要论述了KISS-1基因的结构、组织分布、产物结构和功能,对KISS-2基因的研究进展也作了简单综述.     

The function of the pituitary-testicular axis in dogs prior to and following surgical or chemical castration with the GnRH-agonist deslorelin

Chemical castration, that is the reduction of circulating testosterone concentrations to castrate levels by administration of a GnRH-agonist implant, is a popular alternative to surgical castration in male dogs. Detailed information concerning the pituitary-testicular axis following administration of a GnRH-agonist implant is still scarce. Therefore, GnRH-stimulation tests were performed in male dogs, prior to and after surgical and chemical castration. This approach also allowed us to determine plasma concentrations of testosterone and oestradiol in intact male dogs for future reference and to directly compare the effects of surgical and chemical castration on the pituitary-testicular axis. In intact male dogs (n = 42) of different breeds GnRH administration induced increased plasma LH, FSH, oestradiol and testosterone concentrations. After surgical castration basal and GnRH-induced plasma FSH and LH concentrations increased pronouncedly. Additionally, basal and GnRH-induced plasma oestradiol and testosterone concentrations decreased after surgical castration. After chemical castration, with a slow-release implant containing the GnRH-agonist deslorelin, plasma LH and FSH concentrations were lower than prior to castration and lower compared with the same interval after surgical castration. Consequently, plasma oestradiol and testosterone concentrations were lowered to values similar to those after surgical castration. GnRH administration to the chemically castrated male dogs induced a significant increase in the plasma concentrations of LH, but not of FSH. In conclusion, after administration of the deslorelin implant, the plasma concentrations of oestradiol and testosterone did not differ significantly from the surgically castrated animals. After GnRH-stimulation, none of the dogs went to pre-treatment testosterone levels. However, at the moment of assessment at 4,4 months (mean 133 days ± SEM 4 days), the pituitary gonadotrophs were responsive to GnRH in implanted dogs. The increase of LH, but not of FSH, following GnRH administration indicates a differential regulation of the release of these gonadotrophins, which needs to be considered when GnRH-stimulation tests are performed in implanted dogs.     

促性腺激素释放激素及其类似物在母猪繁殖中应用的研究进展

促性腺激素释放激素(gonadotropin-releasing hormone,GnRH))是下丘脑分泌的生殖激素,主要通过下丘脑-垂体-性腺轴参与调控动物的生殖活动,也可直接作用于性腺或其他器官发挥重要功能。哺乳动物的GnRH具有相同的十肽结构,通过改变十肽结构中第六、九、十位氨基酸可合成不同的GnRH类似物。GnRH及其类似物可通过刺激促黄体素(LH)分泌、抑制雌激素受体二聚化及调节胚胎附植期相关蛋白质的合成来影响动物的繁殖性能。GnRH及其类似物已被证明可提高猪的繁殖力。在母猪生产中,GnRH类似物的应用仍存在受胎次影响、促进排卵但不能增加产仔数等问题。文章主要从GnRH的来源与功能、GnRH及其类似物的结构、GnRH受体(GnRHR)的结构与功能、GnRH及其类似物对母猪繁殖性能的影响,以及存在的问题与展望五方面介绍了GnRH及其类似物在母猪繁殖中的应用研究进展。     

Effect of unilateral castration and unilateral cryptorchidism on gonadotropin and testosterone response to gonadotropin releasing hormone in the bull

The effects of unilateral castration (UC) and induced unilateral cryptorchidism (UCR) on basal plasma luteinizing hormone (LH), follicle stimulating hormone (FSH) and testosterone, and on the responses of these hormones to gonadotropin releasing hormone (GnRH), were investigated in bulls altered at 3, 6 or 9 months of age. Blood plasma was collected before and after GnRH (200 micrograms) stimulation approximately 1 year following gonadal manipulation. Neither mean baseline concentrations nor GnRH-induced increases in plasma testosterone were altered (P greater than .1) by hemicastration or UCR (P greater than .1). Both mean baseline LH and GnRH-induced LH release were greater (P less than .05) in bulls altered at 3 months of age than in bulls altered at 9 months of age. UC increased (P less than .05) plasma LH response to GnRH over that observed in intact bulls, but not above that in UCR bulls. UCR had no detectable effect on either baseline concentrations or GnRH-stimulated LH release. FSH was increased (P less than .05) in hemicastrates, while UCR had a variable effect on peripheral FSH: FSH was reduced (P less than .05) in UCR animals altered at 3 months of age but increased (P less than .05) in UCR bulls altered at both 6 and 9 months of age when compared to FSH in intact bulls. The results indicate that, compared with intact bulls, UC bulls release increased amounts of both gonadotropins but similar amounts of testosterone in response to GnRH stimulation. UCR had a variable effect on FSH release and did not alter either LH or testosterone.     

GnRH及其类似物在动物繁殖中的应用概述

促性腺激素释放激素(GnRH)是由下丘脑神经内分泌小细胞分泌的能促进腺垂体分泌促黄体素(LH)和促卵泡素(FSH)的生殖激素,其类似物较多,在动物繁殖生产实际中也有着广泛的应用。本文阐述了促性腺激素释放激素(GnRH)的化学特性、生理功能及作用特点,同时着重对近年来国内外关于GnRH及其类似物在猪、牛及羊繁殖上的应用研究进行了综述,为更好地应用这一激素提供参考。     

Clinical use of GnRH agonists in canine and feline species

GnRH (gonadotrophin releasing hormone) is a key hormone of reproductive function in mammals; agonist forms have been largely developed, and data concerning their use in small animal reproduction are now abundant. GnRH agonists act by a two-step mechanism. First, their agonist properties on the pituitary will cause marked LH (luteinizing hormone) and FSH (follicle-stimulating hormone) secretion into the bloodstream, accompanied by an increase in the concentrations of sex steroid hormones. Then, in case of constant administration, GnRH agonists will lead to pituitary desensitization, and FSH and LH levels will collapse. These two effects have been widely documented, and these compounds have many potential benefits in a clinical context, capitalizing both on their stimulating and sterilizing effects.     

Relationships of gonadotropins, testosterone, and cortisol in response to GnRH and GnRH antagonist in boars selected for high and low follicle-stimulating hormone levels

Considerable variation exists in the serum levels of gonadotropins in boars; this results in differential testicular function. Boars (Chinese Meishan, European White composite, and crosses of the two breeds) selected for high and low circulating FSH concentrations were used to define possible differences in pituitary sensitivity to GnRH and GnRH antagonist and gonadal and adrenal responses. After a 2-h pretreatment sampling period, boars were injected with GnRH or GnRH antagonist and repetitively sampled via jugular cannula for changes in serum concentrations of FSH, LH, testosterone, and cortisol. In response to varying doses of GnRH or GnRH antagonist, FSH, LH, or testosterone changes were not different in high- or low-FSH boars. Declines in LH after GnRH stimulation were consistently faster in boars selected for high FSH. Chinese Meishan boars had considerably higher cortisol concentrations than White composite boars (132.2 +/- 28.5 vs 67.4 +/- 26.8 ng/mL, respectively; P < .01). When select high- and low-gonadotropin Meishan:White composite crossbreds were sampled, cortisol levels were elevated but comparable between the two groups (126.5 +/- 13.7 vs 131.4 +/- 13.4 ng/mL, respectively). After GnRH antagonist lowered LH concentrations, administration of hCG resulted in increased testosterone and cortisol concentrations. Although testosterone concentrations remained high for 30 h, cortisol concentrations returned to normal levels within 10 h after hCG injection. The mechanism by which boars selected for high gonadotropins achieve increased levels of LH and FSH may not be due to differences in pituitary sensitivity to GnRH but to differences in clearance from the circulation.     

Induced gonadotropin release in adrenocorticotropin-treated bulls and steers

The effect of adrenocorticotropin hormone (ACTH) on plasma cortisol and on gonadotropin releasing hormone (GnRH)-induced release of luteinizing hormone (LH), follicle-stimulating hormone (FSH), and testosterone was determined in nine Holstein bulls and 12 Holstein steers. Treatments consisted of animals receiving either GnRH (200 micrograms, Group G), ACTH (.45 IU/kg BW, Group A) or a combination of ACTH followed 2 h later by GnRH (Group AG). Group G steers and bulls had elevated plasma LH and FSH within .5 h after GnRH injection and plasma testosterone was increased by 1 h after GnRH injection in bulls. In Group A, plasma cortisol was elevated by .5 h after ACTH injection in both steers and bulls, but plasma LH and FSH were unaffected. In Group A bulls, testosterone was reduced after ACTH injection. In Group AG, ACTH caused an immediate increase in plasma cortisol in both steers and bulls, but did not affect the increase in either plasma LH or FSH in response to GnRH in steers. In Group AG bulls, ACTH did not prevent an increase in either plasma LH, FSH or testosterone in response to GnRH compared with basal concentrations. However, magnitude of systemic FSH response was reduced compared with response in Group G bulls, but plasma LH and testosterone were not reduced. The results indicate that ACTH caused an increase in plasma cortisol, but did not adversely affect LH or FSH response to GnRH in steers and bulls. Further, while testosterone was decreased after ACTH alone, neither ACTH nor resulting increased plasma cortisol resulted in decreased testosterone production in the bull after GnRH stimulation.     

Effects of dihydrotestosterone benzoate administration on gonadotropin secretion in ovariectomized pony mares

Eight long-term ovariectomized pony mares were treated with either dihydrotestosterone (DHT) benzoate (400 micrograms/kg body weight) in safflower oil or an equivalent amount of oil every other day for 21 d to determine the effects of DHT on follicle stimulating hormone (FSH) and luteinizing hormone (LH) concentrations in blood samples drawn once daily and after administration of three successive injections of gonadotropin releasing hormone (GnRH). The GnRH injections were given at 4-h intervals on the day following the last DHT or oil injection. Treatment with DHT benzoate did not alter (P greater than .10) concentrations of FSH or LH in daily blood samples relative to controls. The FSH and LH response, assessed by areas under the GnRH curves, decreased (P less than .05) from the first to third injection of GnRH when averaged over both groups of mares. There was no effect of DHT treatment on FSH response to GnRH. There was an interaction (P less than .05) between treatment and GnRH injection for LH areas; areas decreased (P less than .05) for DHT-treated mares from the first to third GnRH injection but were unchanged for control mares. It seems that DHT alone cannot mimic the stimulatory effects of testosterone on FSH production and secretion as observed in previous experiments with ovariectomized and intact mares. Moreover, because intact mares have been shown previously to respond to DHT treatment with an increase in GnRH-induced FSH secretion, it appears that some mechanism is lost in long-term ovariectomized mares, making them unresponsive to DHT treatment.     

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.     

Effects of unilateral castration and unilateral cryptorchidism of the Holstein bull on plasma gonadotropins, testosterone and testis anatomy

The effects of unilateral castration (UC) and induced unilateral cryptorchidism (CR) on plasma hormones and testis anatomy were studied in 36 Holstein bulls altered at either 3, 6 or 9 mo of age (n = 12). Plasma hormone concentrations were determined in six samples collected at hourly intervals on d 0, 1, 3, 7, 14 and 30, and then at monthly intervals through 6 mo after gonadal manipulation. Although plasma testosterone (T) showed a transient decrease (P less than .05) immediately after treatment, mean plasma concentrations of luteinizing hormone (LH) and T were unaffected by UC or CR over the 6-mo period (P greater than .05). Both hormones increased (P less than .05) in concentration with advancing age. Plasma follicle stimulating hormone (FSH) concentration was greater (P less than .05) in UC than in intact (IN) bulls overall, while FSH in CR bulls did not differ (P greater than .05) from either group. At slaughter, 11 mo after gonadal alteration, mean testis weight, ratio of testis weight to body weight and mean testis sperm cell numbers were increased (P less than .05) in UC bulls compared with mean testis values in intact (IN) bulls. Unilateral castration increased (P less than .05) seminiferous tubuler diameter and seminiferous epithelial cell height from basement membrane to the border of the lumen, but did not alter the ratio of tubuler to interstitial space within the testis. Seminiferous tubuler diameter and epithelial cell height were increased (P less than .05) in CR compared with IN bulls. Unilateral gonadal alteration at 3 mo of age caused a greater (P less than .05) hypertrophy of the scrotal testis in both UC and CR bulls than alteration at 6 or 9 mo of age. Results indicate that unilateral gonadal disruption is followed by rapid compensation in testis T production, little change in systemic LH and a rapid increase in secretion of FSH in the bull within those ages investigated. Further, UC elicits a greater compensatory hypertrophy than CR and the pituitary-testis endocrine axis is more responsive to alteration at 3 mo than at 6 or 9 mo of age in the bull.