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。     

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。     

催产素与氯前列烯醇在母猪生产上的应用

1 母猪繁殖周期中激素变化促卵泡素(FSH)是垂体前叶分泌的1种激素,可促进卵巢中卵泡的发育.卵泡成熟时分泌的雌激素会刺激母猪表现出典型的静立发情行为. 促黄体素(LH)刺激卵巢排卵,也是由垂体前叶分泌的.卵泡破裂释放出卵子之后,破裂卵泡内的细胞继续发育,形成黄体. 母猪的正常发情周期是21天,垂体前叶会在发情周期的后期,大约18～20天时分泌FSH.LH是在发情周期的0～2天分泌的,刺激排卵.     

促性腺激素(FSH、LH)对牛腔前卵泡体外发育及E_2分泌的影响

利用McCoy's 5a基础无血清培养系统研究了促卵泡素(FSH)、促黄体素(LH)2种促性腺激素对牛腔前卵泡的体外生长、存活及雌二醇(E2)分泌的影响.结果显示,培养液中单独添加LH对牛腔前卵泡生长影响不大,而单独添加50、100μg/L以上FSH对牛腔前卵泡直径增长和培养后期体外存活有明显的促进作用;当培养液中有10 μg/L LH存在时,2种促性腺激素协同刺激卵泡生长的作用更加明显.添加10 μg/L LH 50μg/L FSH对腔前卵泡体外生长、发育、存活及E2分泌均具有显著的促进作用(P<0.05).     

日粮能量对乌鸡外周血清促性腺激素与垂体分泌促性腺激素的影响

采用体外细胞培养和放射免疫测定法（RIA）测定日粮能量对鸟鸡外周血清促性腺激素与垂体分泌促性腺激素的影响。日粮能量处理：低能Ⅱ组、低能I组、对照组、高能I组、高能Ⅱ组;体外培养垂体细胞组分：高能组、对照纽、低能组。结果显示,血清中FSH含量,对照组与高能Ⅱ组相比差异显著（P〈0．05）,与低能I、Ⅱ组相比差异极显著（P〈0．01）;血清中LH含量,对照组与高能I组、高能Ⅱ组组间差异显著（P〈0．05）;低能Ⅱ组与高能I组、低能Ⅱ组与高能Ⅱ组、低能I组与高能I组、低能I组与高能Ⅱ组组间差异极显著（P〈0．01）;单层垂体细胞中FSH含量,高能组和对照组与低能组比差异显著（P〈0．05）;单层垂体细胞中LH含量,高能组与低能组比较差异极显著（P〈0．01）,对照组与低能组比较差异不显著。结果表明,日粮能量对处理体外培养乌鸡垂体细胞分泌FSH、LH有促进作用。     

促卵泡素和促黄体素对猪卵母细胞体外成熟的影响

本实验探讨了促性腺激素(FSH、LH)对猪卵母细胞体外成熟的影响,并最终找到了其合理的使用剂量.实验选取具有3层致密卵丘细胞层及均匀细胞质的猪卵丘-卵母细胞复合物(COCs)在体外进行成熟培养.体外成熟培养液为改良的TCM-199液,根据实验设计再添加不同浓度的FSH和LH.卵母细胞体外成熟培养条件为39 ℃、5%CO2与饱和湿度下,成熟培养48 h.培养结束后,以第1极体排出作为卵母细胞成熟的标准.在成熟液中添加LH浓度分别为1、5、10 IU/mL时,与没有添加FSH与LH的对照组(成熟率为15.9%)相比,成熟率有显著提高,前者成熟率分别为49.1%、30.0%、36.3%.LH浓度为1 IU/mL的实验组成熟率最高,显著高于对照组(P＜0.05);然后以成熟液中只添加浓度为1 IU/mL LH为对照组,实验组添加FSH浓度分别为1、5、10 IU/mL.结果表明,添加FSH浓度为5 IU/mL和10 IU/mL时其成熟率分别为40.1%、27.6%,低于对照组(49.0%).添加FSH浓度为1 IU/mL时其成熟率(49.1%)高于对照组(49.0%),但差异不显著(P＞0.05).试验表明,随着FSH与LH添加浓度逐渐增加,成熟率呈下降趋势.添加FSH与LH浓度均为1 IU/mL可得到最佳的成熟效果.     

FSH分泌细胞在皖西白鹅脑垂体中的分布与定位

采用免疫组化SABC法并结合DAB显色技术时皖西白鹩脑垂体结构和促卵泡素(follicular stimulating hormone,FSH)分泌细胞进行了研究和定位.结果发现,皖西白鹅的垂体前叶由远侧部和结节部构成,没有中间部.垂体FSH免疫阳性细胞分布广泛,但前叶远侧部分布较多,从而表明皖西白鹅整个垂体都有FSH分泌,主要分泌部位在垂体前叶的远侧部.     

卵母细胞成熟和受精研究的最近进展

本文就培养液中加促性腺激素对于卵母细胞成熟和体外受精的影响进行了探讨。加有LH+E_2、FSH+E_2,及不加激素培养胚胎处理间,合子发育至桑葚胚和囊胚的百分率差异不显著。用不同类型的细胞单层协同培养对于早期胚胎发育的影响试验观察到,在输卵管细胞和子宫细胞组合构成的单层中培养的胚胎比在颗粒细胞单层上培养的胚胎发育更快。有几篇关于分离刺激胚胎发育的输卵管因子的报道认为,在输卵管壶腹部条件培养液中,小鼠1—细胞受精卵发育较好,且证明一种低分子量组分在胚胎发育中起了重要作用。     

双峰驼精清诱导排卵活性成分的DEAE—纤维素柱层析分离及其生物活性

选用DEAE－纤维素离子交换柱对双峰驼精清进行分离，各层析组分用于大鼠垂体组织体外培养，并给母驼肌注有活性的组分，以RIA法测定垂体组织培养液和母驼血浆中LH及FSH的浓度。结果表明，驼精清经DEAE－纤维素柱分离后可得到5个蛋白组分（L1－L5），其中L3在大鼠垂体培养时引起LH释放明显增加（P＜0．05），FSH则无明显变化。给卵巢上无大卵泡发育的母驼肌注有活性的组分L3，可引起血浆中LH明显升高，FSH也无变化。由此表明，L3在体内外试验中均具有引起LH释放的生物活性，它可能是诱导排卵的活性因子或其成分之一。     

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。     

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.     

妊娠期山羊腺垂体中催乳素促卵泡素和促黄体素的分布特点

为了阐明妊娠期垂体中催乳素(PRL)、促卵泡素(FSH),促黄体素(LH)的分布和三者之间协调关系.本试验采用免疫组化SP法检测了PRL、FSH,LH三种激素在妊娠各期山羊的腺垂体细胞中免疫反应产物的分布特点.结果显示,在山羊妊娠过程中,腺垂体中PRL阳性细胞多存在于远侧部,数量逐渐增多,胞质呈强阳性;FSH和LH阳性细胞分布在远侧部和结节部,FSH胞质内颗粒着色深浅不一,LH多核膜着色;相对表达量都逐渐升高,而升高程度不同,表现为妊娠后期LH表达量急剧升高,FSH则变化较小,除了FSH的相对表达量在妊娠前期和中期之间无显著差异(P>0.05)外,3种激素的相对表达量在妊娠前期、中期和后期3个时期之间都存在显著差异(P<0.05),妊娠前、中期FSH的相对表达量均高于PRL和LH,妊娠后期LH的表达量最高.FSH次之,PRL最低.提示PRL,FSH和LH三种激素相互协同以促进胚胎着床并维持妊娠,而且在妊娠早期PRL起主要作用,之后协同FSH和LH维持妊娠.     

Effects of FSH and LH on Steroid Production by Buffalo (Bubalus bubalis) Granulosa Cells Cultured In Vitro Under Serum‐Free Conditions

The objective of this study was to examine the effects of FSH and LH on oestradiol‐17β and progesterone production by buffalo granulosa cells cultured under serum‐free conditions. Granulosa cells (3 × 10⁵) from small (≤5 mm diameter) follicles were cultured for up to 4 days in 48‐well plates coated with 3.3 μg/cm² fibronectin in Dulbecco's modified Eagle's medium (DMEM) : nutrient mixture F‐12 Ham (1 : 1 ratio) supplemented with 10^?7 m androstenedione, 5 μg/ml human apo‐transferrin and 0.1% bovine serum albumin, in the presence or absence of FSH or LH (0, 1, 2, 4, 8, 16, 32 or 64 ng/ml each). Basal oestradiol‐17β production by granulosa cells from small follicles reduced (p < 0.01) from days 1 to 2 of culture and became undetectable by day 3 and basal progesterone production increased (p < 0.05) from day 1 through day 4 of the culture. Although there was no effect of FSH on day 1 of the culture, FSH at 2, 4, 8 and 16 ng/ml increased (p < 0.05) oestradiol‐17β production by granulosa cells from small follicles on day 2. Progesterone secretion was increased (p < 0.05) by all doses of FSH on all days of culture. All doses of LH had no effect on oestradiol‐17β or progesterone production by granulosa cells from small follicles on any day of the culture. The results of this study demonstrate a serum‐free culture system for buffalo granulosa cells and stimulatory effect of FSH but not LH on steroid hormone production by buffalo granulosa cells under these conditions.     

Effect of GnRH and its antagonist (Antarelix) on LH release from cultured bovine anterior pituitary cells

In the following investigations, the LH secretion of cells from pituitaries in heifers on days 16-18 of their oestrous cycle (n = 14) was analysed. Cells were dissociated with trypsin and collagenase and maintained in a static culture system. For the estimation of LH release, the cells were incubated with various concentrations of mammalian GnRH (Lutrelef) for 6 h. To determine the action of Antarelix (GnRH antagonist), the cells were preincubated for 1 h with concentrations of 10(-5) or 10(-4) M Antarelix followed by 10(-6) M GnRH coincubation for a further 6 h. At the end of each incubation, the medium was collected for LH analysis. Parallel, intracellular LH was qualitatively detected by immunocytochemistry. Changes in the intensity of LH staining within the cells in dependence of different GnRH concentrations were not observed, but a significant increase LH secretion in pituitary cells was measured at 10(-6) M GnRH. Antarelix had no effect on basal LH secretion at concentrations of 10(-4) and 10(-5) M. After coincubation of pituitary cells with Antarelix and GnRH, Antarelix blocked the GnRH-stimulated LH secretion with a maximal effect of 10(-4) M, but the staining of immunoreactive intracellular LH was detected at approximately the same level compared to the pituitary cells treated with exogenous GnRH alone. These data demonstrate that Antarelix is effective in influencing the GnRH-stimulated LH secretion of pituitary cells in vitro. After administration of Antarelix in vivo, the GnRH-stimulated LH secretion of cultured pituitary cells was not inhibited.     

Effects of charcoal-extracted follicular fluid on reproductive function in postpartum cows

In order to determine the role of follicle-stimulating hormone (FSH) on the resumption of ovarian function in cows early postpartum (PP), bovine follicular fluid (FF) was used to selectively suppress concentrations of FSH. Calves were removed from all cows within 24 hr of birth. Follicular fluid that was treated with charcoal to remove steroids (15 ml; n = 14) or serum (S) from an ovariectomized cow (15 ml, n = 14) was injected i.m. twice daily from days 1 to 10 PP. Blood samples were collected before each injection and frequent samples (every 15 min for 6 hr) taken on days 5 and 10 PP. Eight cows from each group (FF and S) were slaughtered on the morning of day 11 PP and pituitaries and ovaries collected. The remaining cows (n = 6) were observed for estrus. Treatment with FF delayed follicular growth (P less than 0.01), as evidenced by the largest follicle per cow observed at time of slaughter (3.6 +/- 0.42 vs 11.5 +/- 1.77 mm dia; FF vs S). The intervals from parturition to first estrus (P less than 0.11) and to first progesterone rise (25.3 +/- 1.97 vs 18.0 +/- 3.62 d; P less than 0.06) tended to be delayed by treatment with FF vs S. Many of the cows treated with S ovulated by day 10 PP, they were divided retrospectively into those that had ovulated by (n = 9) or after (n = 5) day 10 PP for analysis. Cows treated with FF had lower (P less than 0.05) and less variable (P less than 0.01) serum FSH concentrations while levels of luteinizing hormone (LH) tended (P less than 0.08) to be greater on days 5 and 10 PP. Follicular fluid decreased levels of FSH (P less than 0.001), but not LH (P less than 0.15), in the samples obtained twice daily compared to S-treated cows that did not ovulate by day 10 PP. Anterior pituitaries were dissociated, and cells from each cow were cultured in order to ascertain whether treatment with FF in vivo would affect gonadotropin secretion in vitro. Estradiol-17 beta (E) was incubated with pituitary cells to determine the effect of E on gonadotropin secretion from cells of PP cows, and to ascertain whether treatment with FF in vivo and with E in vitro would interact to affect secretion of FSH and LH in culture. After 2 d of incubation, cells were treated with 10(-9) M E or vehicle (1% ethanol).(ABSTRACT TRUNCATED AT 400 WORDS)     

Superovulation beim Rind: Hormonprofile bei Stimulation mit Serum-gonadotropin (PMSG) bzw. hypophysärem FSH*)

Contents: Superovulation in cattle: Hormonal profdes during superovulation with PMSG or pituitary FSH.: Hormonal profiles for LH, FSH, PMSG, progesterone in peripheral blood and oestrogens in urine during superovulation with PMSG (1500–3100 IU i. m.) or pituitary porcine FSH (5 mg i.m. 2 times daily for 5 days) were evaluated radioimmunologicaly. Two days after the begin of treatment luteolysis was induced by means of 0.5 mg Estrumat® i. m. The experiments were performed with 14 heifers and 2 cows. Blood samples were taken in 6 or 12 h intervals. Preovulatory LH and FSH peaks occurred coinciding with the onset o f oestrus in good responding animals instead of a few hours later, as in weakly responding animals or during normal oestrous cycle. PMSG could be still measured in peripheral blood 10 days after application. Preovulatory gonadotropin peaks are preceded in good responding animals by clear oestrogen peaks contrary t o weak responding ones. Progesterone concentrations of good responding animals increase faster after the preouulatory LH peak and reach higher absolute values compared t o bad responding ones or during normal oestrous cycle. Furthermore there was a clear difference in progesterone values between both stimulation methods. Progesterone concentrations after induction of superovulation with PMSG are significantly higher f o r about the same number of corpora lutea as after stimulation with pituitary FSH. For judgement of the success of stimulation the determination of progesterone can be considered as parameter. From the hormonal profiles no conclusion can be drawn why some animals don't respond to the stimulation.     

The effect of pulsatile gonadotropin-releasing hormone and estradiol administration on luteinizing hormone and follicle-stimulating hormone concentrations in pituitary stalk-sectioned ovariectomized pony mares

Hourly pulses of gonadotropin-releasing hormone (GnRH) or bi-daily injections of estradiol (E2) can increase luteinizing hormone (LH) secretion in ovariectomized, anestrous pony mares. However, the site (pituitary versus hypothalamus) of positive feedback of estradiol on gonadotropin secretion has not been described in mares. Thus, one of our objectives involved investigating the feedback of estradiol on the pituitary. The second objective consisted of determining if hourly pulses of GnRH could re-establish physiological LH and FSH concentrations after pituitary stalk-section (PSS), and the third objective was to describe the declining time trends of LH and FSH secretion after PSS. During summer months, ovariectomized pony mares were divided into three groups: Group 1 (control, n = 2), Group 2 (pulsatile GnRH (25 μg/hr), n = 3), and Group 3 (estradiol (5 mg/12 hr), n = 3). All mares were stalk-sectioned and treatment begun immediately after stalk-section. Blood samples were collected every 30 min for 8 h on the day before surgery (DO) and 5 d post surgery (D5) to facilitate the comparison of gonadotropin levels before and after pituitary stalk-section. Additionally, jugular blood samples were collected every 12 hr beginning the evening of surgery, allowing for evaluation of the gonadotropin secretory time trends over the 10 d of treatment. On Day 10, animals were euthanized to confirm pituitary stalk-section and to submit tissue for messenger RNA analysis (parallel study). Plasma samples were assayed for LH and FSH by RIA. Mean LH secretion decreased from Day 0 to Day 5 in Groups 1 and 3, whereas LH secretion tended (P < 0.08) to decrease in Group 2 mares. On Day 5, LH was higher (P < 0.01) in Group 2 (17.26 ± 3.68 ng/ml; LSMEANS ± SEM), than either Group 1 (2.65 ± 4.64 ng/ml) or group 3 (4.28 ± 3.68 ng/ml). Group 1 did not differ from Group 3 on Day 5 (P < 0.40). Similarly, mean FSH levels decreased in all groups after surgery, yet Group 2 mares had significantly (P < 0.001) higher FSH concentrations (17.66 ± 1.53 ng/ml) than Group 1 or Group 3 (8.34 ± 1.84 and 7.69 ± 1. 63 ng/ml, respectively). Regression analysis of bi-daily LH and FSH levels indicated that the time trends were not parallel. These findings indicate: 1) Pituitary stalk-section lowered LH and FSH to undetectable levels within 5 d after surgery, 2) pulsatile administration of GnRH (25 μg/hr) maintained LH and FSH secretion, although concentrations tended to be lower than on Day 0, and 3) E2 did not stimulate LH or FSH secretion.     

促卵泡素与促黄体素对猪卵母细胞体外成熟的影响

本试验探讨了促性腺激素(FSH、LH)对猪卵母细胞体外成熟的影响,最终找到了其合理的使用剂量。在成熟液中添加LH浓度分别为1、5、10 IU/mL时,与没有添加FSH与LH的对照组(15.9%)相比,成熟率有显著提高,成熟率分别为49.1%、30.0%、36.3%。LH浓度为1 IU/mL的实验组成熟率最高,显著高于对照组(p<0.05);然后以成熟液中只添加1 IU/mLLH为对照组,试验组分别添加1、5、10 IU/mLFSH。试验结果表明:添加FSH浓度为5和10 IU/ml时其成熟率分别为40.1%、27.6%,低于对照组(49.0%)。添加FSH浓度为1 IU/mL时其成熟率(49.1%)高于对照组(49.0%),但差异不显著(p>0.05)。两组试验同时得出,随着LH与FSH添加浓度的增加,成熟率呈下降趋势。通过本试验结果得出,在体外成熟培养猪卵母细胞时,各添加1 IU/mL LH与FSH时,得到最佳的成熟效果。