大鼠GTH细胞PKC对FSH-β mRNA表达的影响

为研究大鼠促卵泡素(FSH)分泌的受体后信号转导机制,将GTH细胞用PMA或H7处理后,用GnRH脉冲刺激,再用实时荧光定量PCR方法测定细胞FSH-βmRNA表达的Ct值,并与空白对照组比较。结果表明,PMA能显著降低GTH细胞FSH-βmRNA表达的Ct值,H7则曾能显著升高,说明GTH细胞PKC活性显著影响FSH-βmRNA的表达,FSH-βmRNA的表达随着PKC活性的升高而显著升高,随着PKC活性的降低而显著降低。因此,PKC-Ca2+是GnRH脉冲刺激所引起的FSH-βmRNA表达的受体后的信号转导途径。     

促卵泡素和促黄体生成素对儋州鸡生殖激素调控规律的研究

为研究促卵泡素（FSH）和促黄体生成素（LH）对儋州鸡体内其他生殖激素的调控规律,本试验通过改变FSH和LH在儋州鸡血液中的浓度,并采用双抗体一步夹心法酶联免疫吸附试验（ELISA）对处理前后儋州鸡血液中FSH、LH、催乳素（PRL）、孕酮（P）、雌二醇（E₂）的浓度进行测定。结果发现,注射外源性FSH和LH分别能提高儋州鸡血液中FSH和LH浓度;当儋州鸡血液中FSH或LH浓度显著升高时则均能引起PRL浓度显著降低（P < 0.05）,但当FSH和LH浓度同时显著升高时,PRL浓度显著升高（P < 0.05）;当儋州鸡血液中FSH浓度显著升高时,E₂及P浓度显著提升（P < 0.05）,且在高浓度LH的协同下提升幅度更大;当儋州鸡血液中LH浓度显著升高时E₂及P浓度升高但不显著（P > 0.05）。本研究结果表明,儋州鸡血液中FSH或LH浓度的提高均能降低PRL的浓度,并能不同程度的提升E₂及P的浓度,但FSH与LH浓度同时提高则能通过协同作用刺激E₂及P浓度的大幅提升,当E₂及P浓度过高时能通过刺激PRL的释放,负反馈调节血液中FSH与LH,并恢复血液中E₂及P浓度。     

催乳素对鸡胚垂体细胞促性腺激素分泌及生殖相关基因表达的影响

为探讨催乳素(PRL)在垂体层面对禽类生殖的调控机制,以原代培养的鸡胚垂体相关基因表达的变化。结果显示,用5 ng/mL和50 ng/mL的PRL处理鸡胚垂体细胞24 h,对PRLR表达有先抑制后促进作用,对LHβ表达则呈现明显促进作用,对GnRHR和FSHβ表达无显著影响;而对培养液中LH和FSH浓度均无显著影响。当处理时间为12 h时,与对照组相比,50 ng/mL PRL组显著抑制鸡胚垂体细胞分泌LH和FSH;并且在处理12 h后,PRL能抑制LHβ、FSHβ、GnRHR和TGFBR3的表达,而对PRLR和GnIHR的表达水平均无显著影响。研究结果表明,PRL在垂体层面对促性腺激素的分泌和促生殖相关基因表达具有抑制作用。     

细菌类脂多糖(Lps)对桑蚕血球细胞培养系的蛋白激酶C和A活性的诱导

使用含桑蚕血球细胞(主要是颗粒细胞和浆细胞)的离体培养系,经各种刺激物诱导后,对蛋白激酶C(下称PKC)和蛋白激酶A(PKA,需cAMP型)的活性进行了试验。血球细胞经大肠杆菌中的类脂多糖(lipopolysaccharide,LPS)、离子霉素、霍乱弧菌产生的霍乱毒素或笨基乙酸汞(杀菌剂)(4β—phorbor 12—myristate 13 acetate)(PMA)处理后便可清楚地检测到PKC的活性,而没有上述刺激物诱导时,其活性情况没试验过。结果表明,不仅LPS而且Ca~(2+)和一种G蛋白均可能参与了PKC活性诱导的信号转导。同样用LPS、dcAMP、离子霉素或霍乱毒素处理血球细胞,对PKA的活性进行了类似的检测,而不经处理的对照细胞没有PKA活性,而且LPS、cAMP、Ca~(2+)和G蛋白很可能也参与了PKA活性诱导过程中的信号转导。用抗兔脑PKC—α的单克隆抗体进行蛋白质印迹分析,结果以LPS处理的血球细胞样品中有一种与单抗有交叉反应的90KDa蛋白质,而未经LPS处理的对照样品不存在这种蛋白。桑蚕血球细胞内的PKC和PKA被细菌感染后的LPS激活后,可以诱导自身防御反应,例如抗菌蛋白基因的表达。     

褪黑素对体外培养绵羊垂体细胞分泌激素的影响

文章采用体外细胞培养和放射免疫测定(RIA)等方法,研究了褪黑素(MLT)对季节性繁殖的蒙古母羊垂体细胞分泌促卵泡素(FSH)、促黄体素(LH)和促乳素(PRL)的作用。结果表明:当单独用递增的MLT(0、10、100、1000pg/ml)处理原代垂体细胞时,伴随时间的延长FSH的分泌量极显著下降(P<0·01),但对LH的基础分泌没有影响;用10IU/ml hCG单独刺激或用不同剂量MLT与10IU/ml hCG共同刺激垂体细胞,FSH和LH的分泌都极显著高于对照组(P<0·01),但与MLT的剂量没有关系。MLT能改变抑制垂体细胞PRL的分泌,用10IU/ml HCG单独作用使垂体细胞PRL分泌减少,但这种作用可被MLT呈剂量依赖性抑制。本研究表明MLT可作用于季节性繁殖蒙古绵羊的垂体细胞并影响其促性腺激素的分泌。     

初情期前母猪垂体细胞LH和FSH的释放及其对GnRH类似物的反应

利用垂体细胞单层培养模型研究了30，60，90，120和150日龄北京黑猪母猪垂体细胞LH和FSH的释放及其对LRH－A3反应能力。结果表明，猪垂体细胞LH和FSH释放对LRH－A3的反应呈S形剂量依赖型同线。LH基础释放量和LRH－A3刺激的最大释放量在不同日龄间没有显著差异，说明初情期前母猪垂体细胞已达最大LH释放和对GnRH反应的能力。FSH的基础释放量和LRH－A3刺激的最在释放量在30日龄与60日龄之间无差异，但90日龄以后垂体细胞FSH的释施量随日龄增大而降低，提出可能由于垂体在体内时受过抑制素的作用。     

生殖激素的生理功能

1促黄体素 促黄体素（LH）和促卵泡素（FSH）协同作用可促进卵巢血液加速，卵泡最后成熟并分泌雌激素。发情后期，在FSH作用的基础上LH突发性分泌引起成熟卵泡排卵，黄体形成。这种生理机能的出现，孕激素的作用是很重要的。LH通过卵泡壁细胞的环磷酸腺苷（cAMP）促进了孕激素的合成，孕激素则能刺激卵泡壁细胞的蛋白分解酶、胶原水解酶的合成，由于这种酶的出现，分解了卵泡壁细胞而引起排卵。因此，有的动物在排卵前血液中孕激素的含量也相应地升高，排卵后小剂量的LH有利于黄体的维持。     

HMG和FSH一次性注射青年黄牛的药代动力学比较研究

[目的]人绝经期促性腺激素(HMG)是一种具有促卵泡素(FSH)和促黄体素(LH)活性的蛋白质激素,为了比较HMG和FSH一次性注射青年母牛的药代动力学差异和参数。[方法]本研究选择处于发情间期健康状况良好的青年母黄牛20头,随机分成两组,每头牛按4IU/kg体重肌肉注射HMG或FSH,于激素注射后不同时间段采集血样,分离血清后用放射免疫法测定血液中FSH和LH浓度。[结果]表明注射HMG或FSH后血液中FSH与LH水平呈现先升高后下降的趋势,注射HMG后血液中FSH和LH激素水平达峰时间为6h,然后呈缓慢下降趋势,药代半衰期为18h;注射FSH后血液中FSH和LH激素水平在注射后激素血药达峰时间为0.5h,之后呈下降趋势,药代半衰期为6h。单次注射HMG后,0至144h监测期血液中FSH和LH的血药曲线下面积分别为971.21±342.06和176.28±81.80mIU*hr/mL(P0.05),均显著高于单次注射FSH后同期内血液中FSH和LH的血药曲线下面积(分别为39.81±18.01和32.00±20.92mIU*hr/mL,P0.05)。[结论]在青年黄牛体内HMG比FSH的代谢稳定,其活性成分FSH和LH起作用的时间也较长。     

不同日粮能量水平对蛋鸡血液FSH、LH、P4和产蛋率的影响

采用放射免疫分析方法(RIA)测定蛋鸡性成熟过程中和产蛋期血液中FSH、LH和P4的含量,研究不同日粮能量水平对蛋鸡血液FSH、LH、P4和产蛋率的影响,进而研究这些激素与产蛋率的关系。结果表明,高能使见蛋日龄提前,但见蛋后,高能I和低能Ⅱ组产蛋率上升较慢。随着能量的降低,到达产蛋率峰值的时间推迟,峰值以对照组最高,低能组居中,高能组最低。峰值后,高能组产蛋率迅速下降,而其它各组在37周龄前产蛋率维持较高水平的70%左右,到46周龄仍维持在41%以上。在性成熟过程中,血清FSH含量上升,到22周龄时极显著升高,高能组FSH水平显著高于低能组;LH水平也随着周龄的增大而升高;高能I组LH水平显著高于低能组。28周龄高能I组FSH达到峰值,其它各组在33~40周龄达到峰值。高能I、高能Ⅱ和对照组LH水平分别在27、34、30周龄达峰值,低能I和低能Ⅱ组分别在30和42周龄达峰值。在产蛋期,对照组LH水平高于低能组,低于高能组,而对照组产蛋率无一例外地高于其它各组。FSH峰值与平均产蛋率呈显著正相关,产蛋期FSH均值与平均产蛋率呈显著正相关。在性成熟过程中P4含量上升,但18周龄前血清P4含量较低,到22周龄极显著升高;高能组血清中P4水平显著高于低能组。产蛋期,对照和低能组平均P4水平和平均产蛋率均比高能组高。P4峰值和峰后平均值和平均产蛋率呈显著正相关,产蛋期血浆中P4水平与平均产蛋率呈显著正相关。     

垂体促性腺激素表达的受体后信号转导机制

对促性腺激素释放激素(GnRH)作用机制的研究是一个重要领域。文章综述了国内外GnRH的研究进展,认为GnRH与受体(Rs)结合后,要通过一系列的信号转导,调节和控制促卵泡激素(FSH)和黄体生成素(LH)的合成和分泌。GnRH受体后信号转导途径主要是通过3大信号转导系统完成的:环磷酸腺苷(cAMP)信号转导系统、蛋白激酶C(PKC)信号转导系统和丝裂原活化蛋白激酶(MAPK)信号转导系统。cAMP信号转导通路和PKC信号转导通路都属于G蛋白偶联的信号通路,二者最后都要通过激活MAPK信号转导系统来完成调控。这为进一步研究GnRH作用机制提供了依据,同时讨论了存在的问题,并对下一步的研究方向做出展望。     

Luteinizing hormone, follicle stimulating hormone and prolactin secretion in ewes and wethers after zeranol or estradiol injection

Plasma concentrations of luteinizing hormone (LH), follicle stimulating hormone (FSH) and prolactin (PRL) were determined over a 24-h period using radioimmunoassay in sheep injected with corn oil (control) or various doses of zeranol or estradiol-17 beta. Injection of .333, 1 or 10 mg of zeranol caused dose-related increases (P less than .01) in plasma PRL (peak levels at 12 to 18 h) and LH (peak levels at 12 to 20 h) in ovariectomized ewes. Similarly, PRL and LH increased following doses of 33 or 100 microgram of estradiol. Before the LH surge, plasma LH levels were significantly depressed (4 to 8 h). Plasma FSH levels were significantly decreased 4 to 8 h after zeranol and estradiol injection. Slight surges of FSH were observed at times similar to those of LH, but the peak level was never greater than control levels. Injection of 1 mg of zeranol or 100 microgram of estradiol into wethers resulted in a 24-h pattern of PRL secretion not significantly different of LH concentration and significantly prolonged inhibition of FSH secretion. These results indicate similarities in the effects of zeranol and estradiol on anterior pituitary hormone secretion within groups of animals of the same sex or reproductive state. Differences in secretion and plasma concentrations of LH, FSH and PRL due to underlying sexual dimorphism are maintained and expressed even when animals are challenged with structurally different compounds of varying estrogenic potencies.     

Effects of adding luteinizing hormone to a medium containing follicle stimulating hormone on progesterone-induced differentiation of cumulus cells during meiotic resumption of porcine oocytes

In the present study, we investigated the effects of adding luteinizing hormone (LH) to a medium containing follicle stimulating hormone (FSH) on the shift in expression of progesterone receptor (PR) isoforms (PR‐A and PR‐B) and the roles in function of cumulus cells of cumulus‐oocyte complexes (COC). The level of PR‐B mRNA in cumulus cells was up‐regulated by FSH during the first 16‐h cultivation but the level was significantly decreased at 20 h. The decrease of PR‐B mRNA was accelerated when COC were cultured with FSH and LH. Still, a high level of total PR mRNA was maintained in cumulus cells cultured with or without the addition of LH up to 20 h, suggesting that the expression of PR isoforms was shifted from PR‐B to PR‐A in cumulus cells. The reduction of PR‐B was also induced by addition of progesterone to FSH‐containing medium. The addition of LH or progesterone to FSH‐containing medium stimulated cumulus expansion of COC as compared with that of COC cultured with FSH. In the expanded COC, ADAMTS‐1 which is expressed in granulosa cells and cumulus cells in rodent follicles through LH‐induced progesterone‐ and PR‐dependent pathway, was more accumulated within the COC matrix. These results suggest that the addition of LH or progesterone to FSH‐containing medium is required for the differentiation of cumulus cells, such as cumulus expansion, mediated by the shift from PR‐B to PR‐A in them.     

Changes in the pituitary-gonadal axis associated with puberty in Holstein bulls

The temporal pattern of the endocrine changes associated with puberty were studied using 52 bulls born in October or April. Blood samples were taken weekly and at 30-min intervals for 5 h every 4-wk. Bulls were castrated at one of six 4-wk intervals between 12 and 32 wk and blood samples were taken. Season of birth affected concentrations of testosterone (greater for spring-born) in intact bulls, but not luteinizing hormone (LH) or follicle stimulating hormone (FSH). The concentration of FSH increased about 30% between 4 and 32 wk, without evidence of pulsatile discharge. Basal concentration of LH was low and pulsatile discharges were infrequent at 4 or 8 wk. At 12, 16 and 20 wk, however, basal LH concentration was elevated and LH discharges were at less than 2-h intervals. Testosterone concentration did not rise until 18 to 20 wk, but then continued to rise; LH discharge was suppressed concomitantly. Bulls castrated at 16 or 20 wk had higher concentrations of LH in their blood both before and shortly after castration values for bulls, but by 21 d after castration values for bulls of all ages were similar. It was concluded that elimination of an unidentified suppressive factor allows frequent discharges of LH between 12 an 16 wk, but the testes do not respond by secreting more testosterone until 18 to 20 wk. By 24 wk, the testes are secreting more testosterone and pituitary production of LH is restored to a lower level; LH discharges decline in frequency and basal LH level declines. The high frequency discharges of LH between 12 and 20 wk are postulated to induce responsiveness of Leydig cells to LH and, thus, enable elevation of intratesticular testosterone to levels necessary for Sertoli cell differentiation and initiation of spermatogenesis.     

Pituitary and testicular responses of beef bulls to active immunization against inhibin alpha

Prepubertal crossbred beef bulls served as controls or were actively immunized against the N-terminal, 30-amino acid synthetic fragment of porcine inhibin alpha, pI alpha (1-30). Antibody titers were detected in sera (greater than 40% B/BO in sera diluted 1,000-fold) but not in rete testis fluid of 390-d-old bulls. Serum FSH and inhibin remained static during a 5-h intensive bleed; inhibin was not acutely affected by a 15-fold LH rise and a threefold FSH rise induced by exogenous GnRH. Serum FSH, but not LH or testosterone, was consistently elevated (P less than .05) in immunized bulls compared with control bulls. Neither pituitary weight, pituitary gonadotropin content nor pituitary FSH/LH ratios were affected (P greater than .10) by pI alpha(1-30) active immunization. Testicular sperm density was greater (60 x 10(6) vs 45 x 10(6) sperm/g testis; P less than .10) in immunized bulls, but testes weight, epididymides weight and total daily sperm production remained unchanged. These results suggest that inhibin is important for regulation of FSH secretion and testicular function. Immunization with suitable inhibin vaccines may improve bull fertility.     

α-Adrenergic control of gonadotropin secretion in the ewe

The effect of the centrally acting α-adrenoceptor agonist, clonidine, on plasma LH and FSH was studied in oestradiol-primed and unprimed ewes and in oestrous ewes. In unprimed anoestrous ewes, clonidine stimulated LH and FSH release after a lag period of 18 h, and noradrenaline intracarotid injection or i.v. infusions immediately stimulated LH release. In oestradiol-infused anoestrous ewes, clonidine produced either a delay or inhibition of the gonadotrophin surge and noradrenaline i.v. infusion advanced the LH surge. In oestrous ewes treated with clonidine, there was marked delay in the LH surge, but the magnitude of the LH and FSH surges were unaffected. Intravenous administration of α-adrenoceptor blockers, phentolamine and phenoxybenzamine, blocked the oestradiol-induced gondotrophin surge in anoestrous ewes. The effect of phenoxybenzamine on gonadotrophin surge was dose dependent in oestrous ewes. Small doses (4 mg/kg i.v.) of phenoxybenzamine delayed the synchronous LH and FSH surges. There was complete blockade of the LH surge and partial blockade of FSH surges in ewes given phenoxybenzamine (8 mg/kg i.v.) before the expected synchronous gonadotrophin surges. After this experiment, the initial rise of plasma progesterone concentrations did not occur until day 6 of oestrous cycle. Administration of phenoxybenzamine before the expected second FSH surge had no effect on the second FSH surge. Gonadotrophin release induced by gonadotrophin-releasing hormone was attenuated by phenoxybenzamine, but not by clonidine. The results suggest that the LH surge is under α-adrenergic control and the first FSH surge is under partial α-adrenergic control, but the second FSH surge is not under α-adrenergic control. The results also suggest oestradiol modulation of α-adrenergic receptor action.     

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.     

Study on Regulation of Reproductive Hormones in Danzhou Chicken by FSH and LH

To investigate the regulation of reproductive hormones in Danzhou chicken by follicle-stimulating hormone (FSH) and luteotropic hormone (LH),the concentration of FSH and LH in Danzhou chicken blood were changed by different treatments and the concentration of FSH,LH, prolactin (PRL), progesterone (P), and estradiol (E₂) in the blood were determined using the enzyme-linked immunosorbent assay (ELISA).The results showed that exogenous FSH and LH could improve the concentrations of FSH and LH,respectively. When one of FSH and LH increased alone,the concentration of PRL was significantly decreased (P < 0.05),while it was significantly increased when FSH and LH were both increased (P < 0.05).The high level of the FSH could result the significant increase of E₂ and P (P < 0.05),and it would expand the increasing effect in cooperation with the high concentration of LH. While the high level of the LH could elevate the concentration of E₂ and P,but effect was not significant (P > 0.05).In conclusion,the study suggested that the increase level of FSH or LH could reduce the concentration of PRL in Danzhou chicken,meanwhile,it could increase the concentration of E₂ and P in varying degrees. However,both of the FSH and LH increase in the blood could result the significant increase of E₂ and P as the synergy. When the concentration of E₂ and P were too high,it could stimulate the release of PRL which could adjust the concentration of FSH and LH,and rebalance the concentrations of E₂ and P in the blood.