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。     

不同月份产羔母羊泌乳期生殖激素与生长激素监测

为了解不同月份生产的奶山羊泌乳期生殖激素与生长激素(GH)变化规律,随机抽取1、3、5和8月份分娩的奶山羊各10只,于分娩后0 d、7 d、1~8个月(每月的第15天)时采集母羊静脉血,分离血清,采用ELISA试剂盒检测母羊外周血中催乳素(PRL)、卵泡刺激素(FSH)、促黄体生成素(LH)、雌激素(E₂)、孕酮(P₄)和GH水平变化。结果显示:不同月份产羔母羊泌乳期间外周血中同一激素的动态变化趋势一致;在整个泌乳期间,1、3、5、8月份产羔母羊PRL、FSH、LH、P₄、E₂、GH含量的动态变化范围分别为446.17~221.72 ng·L^-1、9.49~3.82 U·L^-1、351.17~218.16 pg·mL^-1、4086.83~3568.15 pmol·L^-1,33.74~22.30 ng·L^-1、30.36~11.57μg·L^-1;不同月份产羔母羊外周血中GH水平在相同泌乳期均无显著差异(P>0.05),FSH水平在0 d有显著差异(P<0.05),P₄水平在8个月时有显著差异(P<0.05),PRL水平在泌乳的0 d和2、3、8个月时有显著差异(P<0.05),LH水平在泌乳的0 d、2个月时有显著差异(P<0.05),E₂水平在泌乳0 d、3个月和8个月时有显著差异(P<0.05)。结果表明:奶山羊产羔月份的不同对泌乳期间的激素水平有一定的影响,但动态变化趋势一致。     

催乳激素对山羊脂肪酸合酶基因转录活性的影响

为探讨催乳激素对体外培养的山羊乳腺上皮细胞脂肪酸合酶（fatty acid synthase，FASN）基因转录活性的调控作用，试验分别用不同浓度的胰岛素（insulin，INS）、雌激素（estradiol，E₂）、催乳素（prolactin，PRL）及不同激素组合（INS+PRL、E₂+INS+PRL）处理山羊乳腺上皮细胞24 h，提取细胞总RNA，采用实时荧光定量PCR检测催乳激素对FASN基因mRNA表达水平的影响。细胞转染山羊FASN基因启动子报告基因载体，同样用不同浓度的胰岛素、雌激素、催乳素及激素组合处理24 h，利用双荧光素酶报告基因系统检测催乳激素对FASN基因启动子活性的影响。结果发现，用雌激素、催乳素处理山羊乳腺上皮细胞后，FASN基因启动子活性及mRNA水平极显著或显著上调（P<0.01；P<0.05），雌激素浓度为10、100 μmol/L和催乳素浓度为0.1和1 μg/mL时效果最为明显，而胰岛素对FASN基因的启动子活性及mRNA水平没有显著影响（P>0.05）。用不同激素组合（INS+PRL、E₂+INS+PRL）处理细胞均能显著上调FASN基因启动子活性及mRNA表达水平（P<0.05）。结果表明，雌激素和催乳素能够调控FASN基因的转录活性，为进一步研究泌乳过程中FASN基因的分子调控机制提供理论依据。     

黄体期注射PGF2α对母羊血清生殖激素及相关细胞因子的影响

旨在研究黄体期不同阶段注射前列腺激素(PGF2α)对育成母羊生殖激素和生殖相关细胞因子的影响。本研究选择健康、体况良好、体重相近、发情周期正常的湖羊育成母羊60只,用“孕酮栓(MAP)+PMSG”法进行发情周期同步化处理后,选择发情正常的48只母羊随机均分为6组。发情当天记为第0天,黄体前期试验组、中期试验组和末期试验组母羊分别在第6(黄体前期)、11(黄体中期)、16天(黄体末期)注射1 mL PGF2α(0.1 mg),黄体前期对照组、中期对照组和末期对照组母羊分别在第6、11、16天注射1 mL生理盐水,每次注射后0.5、1、2、3 h采血,用于血液指标检测。结果表明,所有试验组和对照组母羊于注射后的0.5~3 h间血清中FSH、LH、PRL、P₄、E₂水平以及TNF-α、IL-1β、IL-6、IFN-β无显著变化(P＞0.05);母羊在黄体期不同阶段注射PGF2α对0.5、1、2、3 h血清中FSH、LH、PRL及IL-1β、IFN-β无显著影响(P＞0.05),前期试验组注射后3 h P₄水平显著低于前期对照组,E₂和IL-6水平显著高于前期对照组(P＜0.05),前期试验组注射后2和3 h TNF-α水平显著高于前期对照组(P＜0.05);中期试验组注射后1 h P₄水平显著低于中期对照组(P＜0.05)。黄体前期注射PGF2α后,前期试验组0.5~ 3 h内FSH、E₂、TNF-α、IL-6整体水平显著高于前期对照组(P＜0.05),P₄整体水平显著低于前期对照组(P＜0.05),对LH、PRL、IL-1β和IFN-β无显著影响(P＞0.05);黄体中期注射PGF2α后,中期试验组0.5~3 h内E₂整体水平显著高于中期对照组(P＜0.05),P₄整体水平显著低于中期对照组(P＜0.05),对FSH、LH、PRL、TNF-α、IL-1β、IL-6和IFN-β无显著影响(P＞0.05);黄体末期注射PGF2α对生殖激素与相关细胞因子没有显著性影响(P＞0.05)。本研究结果表明,PGF2α对黄体的溶解作用存在阶段性差异,母羊在黄体前期对PGF2α的短期应答反应强于黄体中、末期,且黄体前期时,卵巢可以响应PGF2α为卵泡发育营造更佳的发育环境。     

BMP4诱导的Smad9在小鼠卵泡发育中的作用及其机制的初步研究

本研究旨在探讨Smad9在小鼠卵泡发育中的作用，为卵泡生长发育及其调控机制的研究提供新思路。将6周龄的雌性昆明小鼠随机分成3组，分别腹腔注射生理盐水、骨形态发生蛋白-4（BMP4）和Smad9抑制剂（LDN-193189），依次作为对照组、BMP4组和LDN组。48 h后收集卵巢制作石蜡切片，HE染色后观察卵泡发育情况并对各级卵泡进行计数；采用Western blotting和实时荧光定量PCR技术检测Smad9蛋白和mRNA水平。同时通过ELISA试验测定各组小鼠血清中雌二醇（E₂）、孕酮（P₄）、黄体生成素（LH）、卵泡刺激素（FSH）和芳香化酶等的浓度，并检测了CYP19a1、LHR、PRLR和FSHR基因的表达情况，进一步探究Smad9可能的作用机制。结果显示，与对照组相比，BMP4组Smad9蛋白和mRNA水平较高，而LDN组较低；BMP4组有腔卵泡显著增加（P<0.05），而LDN组显著减少（P<0.05）。此外，ELISA试验结果显示，BMP4组血清E₂、FSH和芳香化酶含量增加，而P₄和LH含量降低；LDN组中E₂和芳香化酶的含量降低。实时荧光定量PCR结果显示，BMP4组FSHR和CYP19a1基因的mRNA水平升高，而LHR和PRLR基因的mRNA水平降低；LDN组PRLR基因的mRNA水平升高，而CYP19a1基因的mRNA水平降低。由以上试验结果可以得出，BMP4诱导的Smad9能促进小鼠有腔卵泡的发育，而其可能主要通过影响E₂、PRL和芳香化酶等的产生而起作用。     

Opioid and 17β-estradiol regulation of LH and FSH secretion during sexual maturation in heifers

The objective of the present study was to examine the involvement of opioid neuropeptides and E₂ in regulating circulating concentrations of gonadotropins during sexual maturation in the bovine female. Prepubertal (immature) and postpubertal (mature) bovine females were used. Mean concentrations of luteinizing hormone (LH) and follicle- stimulating hormone (FSH) in circulation before and after administration of naloxone were determined in ovariectomized heifers administered E₂ and ovariectomized heifers not administered E₂. A linear decline (P<0.01) in opioid suppression of LH and FSH occurred during the experimental period in immature heifers receiving E₂. This decline in opioid suppression of LH and FSH occurred during the same period of time that intact control heifers were initiating estrous cycles at puberty. Little change of opioid suppression of LH and FSH occurred during the experimental period in immature heifers not receiving E₂ and mature heifers receiving E₂. Our research indicates that opioid neuropeptides and E₂ act together to regulate LH and FSH secretion during sexual maturation in the bovine female.     

Circulating concentrations of 17-estradiol influence pattern of LH in circulation of cows

The objective of the research was to determine the relationship between circulating 17β-estradiol (E2) and secretion of luteinizing hormone (LH) in cows. A second objective was to determine if response to E₂ was influenced by interval between ovariectomy and the start of E₂ treatment. Thirty-one nulliparous cows 3 yr of age were randomly assigned to a 2 × 4 factorial arrangement of treatments. Sixteen cows were ovariectomized at 18 mo of age (long term), and the other 15 cows were ovariectomized at 36 mo of age (short term). At the time of ovariectomy of cows in the short term group, 11 cows in the short term group and 12 cows in the long term group were implanted subcutaneously with 1, 2 or 4 polydimethylsiloxane capsules containing E₂. The other eight cows served as non-implanted controls (n=4-short term, n=4-long term). All cows were fitted with jugular vein catheters on day 29 of treatment, and on day 30 blood samples were collected at 12-min intervals for 6 hr. At the end of 6 hr, luteinizing hormone-releasing hormone (LHRH) was administered and blood sampling continued at 12-min intervals for an additional hour. Serum was analyzed for LH and E₂. Variables of LH secretion analyzed were mean concentration, frequency of pulses, amplitude of pulses and maximum concentration after LHRH. There were no significant interactions for any of the variables of LH among cows ovariectomized for the long and short term. There was a significant linear increase in mean concentration of LH with increased circulating concentration of E₂. Frequency of LH pulses was not affected by circulating concentration of E₂. As circulating concentration of E₂ increased, amplitude of LH pulses increased and response to LHRH increased - resulting in an increase in mean LH. Interval from time of ovariectomy to the start of E₂ treatment only had a minor influence on mean concentration of LH and profile of LH concentrations in circulation.     

人尿促性腺激素、雌二醇及表皮生长因子对牛卵母细胞体外成熟质量的影响

试验旨在研究不同激素配比及表皮生长因子(EGF)浓度对牛卵母细胞体外成熟及卵母细胞质量的影响。将随机分组的卵丘-卵母细胞复合体于添加FSH+LH、HMG、FSH+LH+E₂、HMG+E₂ 4种不同激素组合配比的成熟基础液中培养,对比其体外成熟率,比较了EGF对牛卵母细胞体外成熟率和孤雌胚胎体外发育的影响,并采用TUNEL法检测添加不同浓度EGF的牛孤雌激活囊胚细胞凋亡情况。结果表明,添加HMG的成熟试验结果稳定,E₂对牛卵母细胞成熟有一定的促进作用,HMG+E₂联合使用可以得到高效稳定的成熟结果;在此基础上,在成熟液中添加30 ng/mL EGF对牛卵母细胞的成熟质量、胚胎发育及降低胚胎细胞凋亡都有明显的促进作用。因此,在体外成熟培养液中添加0.075 IU/mL HMG、1 μg/mL E₂和30 ng/mL EGF对牛卵母细胞的成熟和质量较为有益。     

Effect of Human Menopausal Gonadotropin,17β-estradiol and Epidermal Growth Factor on the Quality of in vitro Maturation Bovine Oocytes

For optimizing in vitro maturation system of bovine oocytes,we firstly examined the influence of four different hormonal regimes(FSH+LH,HMG,FSH+LH+E₂ and HMG+E₂) on oocyte maturation rates.Then we studied the effects of epidermal growth factor (EGF) in the above defined medium on bovine oocyte maturation,in vitro development and quality of parthenogenetic embryos.The cell apoptotic index of parthenogenetic blastocysts was detected by TUNEL.No significant difference was observed in maturation rates in four groups supplemented with different hormones.However,human menopausal gonadotropin (HMG) provided steady maturation results in replicates.Maturation of oocytes was promoted by supplementation with 17β-estradiol (E₂).Combination of HMG and E₂ gave rise to steady and efficient mature results.The presence of EGF at 30 ng/mL concentration significantly increased maturation rate and blastocyst rate and reduced apoptotic cells in parthenogenetic blastocysts.Therefore,the optimal oocyte maturation solution could be supplemented with 0.075 IU/mL HMG,1 μg/mL E₂ and 30 ng/mL EGF.     

小花棘豆中毒对和田羊母羊繁殖器官的影响

试验旨在探讨小花棘豆（Oxytropis glabra DC）中毒对和田羊母羊繁殖器官指数、性激素水平和相关基因mRNA表达量的影响。以小花棘豆中毒和田羊母羊为研究对象,采集血清后屠宰,采集试验羊的丘脑、垂体和卵巢组织,测定其脏器系数,检测血清中促性腺激素释放激素（GnRH）、促卵泡素（FSH）、促黄体素（LH）、雌二醇（E₂）和孕酮（P₄）含量的变化,并检测各组织中相关繁殖基因的表达。结果显示,小花棘豆中毒和田羊母羊丘脑、垂体和卵巢指数均极显著升高（P < 0.01）,且卵巢表面卵泡数显著下降（P < 0.05）,HE染色表明丘脑神经元细胞固缩、浓染;垂体中细胞核变形、浓染,胞浆减少;卵巢中初级卵母细胞溶解、消失,间质血管扩张。小花棘豆中毒和田羊母羊血清中GnRH、FSH、LH、E₂和P₄含量均极显著下降（P < 0.01）,丘脑中Kiss-1、GPR54、ERα mRNA,垂体中GnRHR mRNA和卵巢中FSHR、LHR mRNA表达量均极显著下降（P < 0.01）。结果表明,小花棘豆毒性成分可通过丘脑-垂体-性腺轴影响和田羊母羊的生殖系统。     

饲粮钙磷水平对伊犁马初乳成分、乳脂脂肪酸组成、产后血液钙磷及激素的影响

旨在研究妊娠后期饲喂不同钙磷水平饲粮对产后伊犁马初乳成分、乳脂脂肪酸组成、血液钙磷及钙磷代谢激素、繁殖及生长代谢激素等生理生化指标的影响，为明确伊犁马在妊娠后期对钙磷的适宜需要量提供参考。本试验选取健康、无亲缘关系、12~13周岁、体重为（380±32）kg、胎次为4~5胎、处于第10妊娠月左右的伊犁马25匹，随机分为5组，每组5个重复。各组饲粮钙饲喂水平分别为36.00、39.00、42.00、45.00和48.00 g·d^-1，磷饲喂水平分别为26.30、28.30、30.30、32.30和34.30 g·d^-1。试验预试期10 d，正试期从第11天至母马分娩后第2天结束。母马分娩后12 h内采集初乳用于测定乳成分和乳脂脂肪酸含量，空腹血液用于测定Ca、P、甲状旁腺素（PTH）、降钙素（CT）、骨钙素（OC）、胎盘催乳素（PL）、垂体催乳素（PRL）、雌酮（E₁）、雌二醇（E₂）、血清孕酮（PROG）、甲状腺素（T₄）、三碘甲状腺原氨酸（T₃）、生长激素（GH）、胰岛素样生长因子-Ⅰ（IGF-1）等激素浓度。结果表明：1）饲粮钙磷水平对初乳中钙、磷、乳脂率、乳糖率均无显著影响，组间乳蛋白率、总固形物、体细胞数、乳脂中饱和脂肪酸（SFA）和不饱和脂肪酸（UFA）比例存在显著差异（P<0.05）。随着饲粮钙、磷水平的增加，乳磷浓度、乳蛋白率和UFA比例呈显著线性增加（P<0.05），总固形物呈显著二次增加（P<0.05），而乳糖率、SFA及肉豆蔻酸比例呈显著线性降低（P<0.05）。2）饲粮钙磷水平显著影响血液中OC和PROG浓度（P<0.05），随着饲粮钙磷水平的增加，血液中离子钙、CT和OC浓度呈显著的线性降低（P<0.05），而PTH和PROG水平呈显著的线性升高（P<0.05）。饲粮钙、磷水平对PL、PRL、E₁、E₂等生殖激素以及T₃、T₄、GH和IGF-1等生长代谢激素浓度均无显著影响。由此可见，在本试验条件下，饲粮钙磷通过提高初乳中乳蛋白率和总固形物来改变乳脂中饱和脂肪酸含量和不饱和脂肪酸比例影响初乳成分；通过降低血液中OC浓度影响钙磷代谢；通过提高血液中PROG浓度影响繁殖状况。     

定时输精对经产母猪繁殖性能及生殖激素的影响

试验旨在建立高效的经产母猪定时输精（timed artificial insemination,TAI）技术,研究了定时输精对经产母猪繁殖性能、断奶-分娩间隔、不同胎次母猪产仔性能及断奶后7 d内血清生殖激素水平的影响。选取309头2~8胎次二元（长×大）经产母猪,随机分为对照组和试验组,对照组母猪进行常规人工授精（artificial insemination,AI）,试验组母猪进行断奶后24 h注射PMSG 1 000 IU,间隔72 h注射GnRH 100 μg,在注射GnRH后24和40 h各输精1次的定时输精技术。通过统计两组母猪的断奶1周内发情率、受胎率、分娩率、窝均产仔数等,判断定时输精对经产母猪繁殖性能的影响;通过对断奶时间和分娩时间的统计,检测定时输精对经产母猪断奶-分娩间隔的影响;用放射免疫（RIA）方法检测2~4胎次母猪断奶1周内血清E₂、LH、FSH和P₄的含量,研究定时输精对母猪生殖激素的影响。结果显示,试验组母猪发情率显著高于对照组（P<0.05）,但两组间受胎率、分娩率差异不显著（P>0.05）,窝均产仔数、窝均合格仔数和繁殖效率有增加的趋势,但差异不显著（P>0.05）;定时输精显著缩短了母猪的断奶-分娩间隔（P<0.05）。在胎次方面,3~4胎母猪使用定时输精的效果较好,其发情率、受胎率和分娩率均显著高于对照组（P<0.05）。在生殖激素方面,试验组E₂水平在注射PMSG后迅速上升,且在定时输精处理后66~96 h内持续高于对照组（P<0.05）,试验组P₄水平在断奶后至配种前显著低于对照组（P<0.05）,但配种后快速升高,并高于对照组;LH和FSH的含量在两组间无显著差异。综上,定时输精可有效提高经产母猪的断奶发情率,并减少其非生产天数,可显著提高3~4胎母猪的繁殖性能。     

Effects of Oxytropis glabra DC Poisoning on the Reproductive Organs in Hetian Sheep Ewes

The study was aimed to research the effects of Oxytropis glabra DC (O.glabra DC) poisoning on reproductive organs coefficent, reproductive performance and related gene expression in Hetian sheep ewes. The sera, hypothalamus, pituitary and ovary of Hetian sheep ewes cases poisoning of O. glabra DC were collected, the organ exponent and contents of GnRH, FSH, LH, E₂ and P₄ in serum were measured, then the mRNA expression of reproductive genes were measured. The result demonstrated that the index of hypothalamus, pituitary and ovary were extremely significantly higher than normal sheep (P < 0.01),the follicle number on the surface of the ovaries in poisoning groups were significantly lower than normal sheep (P < 0.05). The result of hematoxylin-eosin staining showed that the nuclear of neuronal cells in hypothalamus were pyknosis and hyperchromatic; The nuclear of cells in pituitary deformation and hyperchromatic, cytoplasm decreased; The primary oocyte in ovary dissolved, disappeared, interstitial blood vessel expanded. The content of GnRH, FSH, LH, E₂ and P₄ in serum in poisoned Hetian sheep ewes were extremely significantly lower than normal sheep (P < 0.01). The mRNA expression levels of Kiss-1, GPR54, ERα in hypothalamus, GnRHR in pituitary and FSHR and LHR in ovary were extremely significantly lower than normal sheep (P < 0.01). The result showed that O. glabra DC poisoning could affect Hetian sheep ewes reproductive system by affecting microstructure and ultrastructure of hypothalamus-pituitary-ovarian axis.     

日粮粗蛋白质水平对伊犁鹅产蛋规律、繁殖激素分泌及相关基因mRNA表达的影响

为探讨日粮粗蛋白质水平对伊犁鹅的产蛋规律、繁殖激素分泌及生殖轴相关基因mRNA表达量的影响,本研究随机选取200只3岁伊犁鹅（年龄、饲养管理水平一致及体重相近）,随机分为4个组,每组10个重复。对照组及公鹅饲喂鹅场自配料（粗蛋白质水平11.20%）,试验组母鹅分别饲喂粗蛋白质水平为13.86%、15.20%和16.48%的日粮,其余营养指标基本一致。预试期1周,正试期9周。结果表明：①试验期内伊犁鹅的产蛋率呈波动变化,对照组伊犁鹅的产蛋率在第1~2周逐渐上升,第4~5周出现小幅度增长,其余阶段表现为下滑趋势;各试验组伊犁鹅的产蛋率在第1~2、4~5周出现较大幅度增长,且在第5周时达到产蛋高峰,此后逐渐下滑。15.20%粗蛋白质组伊犁鹅就巢率显著低于11.20%、13.86%粗蛋白质组。②与对照组相比,13.86%粗蛋白质组血清促性腺激素释放激素（GnRH）浓度极显著升高（P<0.01）,且血清雌二醇（E₂）、促黄体素（LH）浓度均显著升高（P<0.05）;15.20%、16.48%粗蛋白质组血清GnRH、E₂及LH浓度均极显著升高（P<0.01）,且血清促卵泡素（FSH）浓度显著升高（P<0.05）。各试验组血清催乳素（PRL）浓度均显著降低（P<0.05）。③与对照组相比,15.20%粗蛋白质组下丘脑GnRH、LHR基因表达量显著上调（P<0.05）,PRL、PRLR基因表达量显著下调（P<0.05）;垂体中LH基因表达量显著上调（P<0.05）;卵巢中LH与ESR2基因表达量显著或极显著上调（P<0.05,P<0.01）。综上所述,基于日粮粗蛋白质水平对伊犁鹅的产蛋率、就巢率、血清生殖激素浓度及生殖轴相关基因mRNA相对表达量影响的综合评估,建议产蛋期伊犁鹅日粮粗蛋白质水平为15.20%。     

褪黑素对荷斯坦奶牛妊娠率及生殖激素的影响

试验旨在探明皮下注射褪黑素（MT）对荷斯坦奶牛配种妊娠率及血清生殖激素的影响。用计步器法确定自然发情的首次配种荷斯坦奶牛150头，对其中70头进行颈部皮下肌内注射褪黑素30 mg，12 h后进行人工输精；选择170头产后首次配种的荷斯坦奶牛进行同期排卵-定时输精处理，其中90头荷斯坦奶牛最后一次注射促性腺激素释放激素（gonadotropin-releasing hormone，GnRH）的同时进行颈部皮下肌内注射褪黑素30 mg，16 h后进行人工输精。在进行二次配种的荷斯坦奶牛中选择153只进行皮下注射褪黑素。荷斯坦奶牛输精后20~35 d进行妊娠检查，详细记录首次配种妊娠母牛头数、二次配种妊娠母牛头数、产犊数。选取同期排卵-定时输精的荷斯坦奶牛25头，皮下注射褪黑素8 h后用放射免疫法检测其血清中褪黑素、促卵泡素（FSH）、促黄体素（LH）、雌二醇（E₂）的含量及35 d妊检时妊娠母牛血清中孕酮（P₄）含量。结果显示，与自然发情和同期排卵对照组相比，其对应的皮下注射褪黑素组荷斯坦奶牛的妊娠率及产犊率均显著提高（P<0.05）；皮下注射褪黑素组的双犊率显著提高（P<0.05），首次配种妊娠率和产犊率均显著提高（P<0.05），二次配种妊娠率和产犊率均差异不显著（P>0.05）。血清激素检测结果表明，与对照组相比，皮下注射褪黑素组血清中褪黑素、LH、E₂含量均显著增加（P<0.05）；35 d妊检时，皮下注射褪黑素的妊娠母牛P₄含量显著增加（P<0.05）。本试验结果表明，皮下注射褪黑素能够提高荷斯坦奶牛的妊娠率、产犊率及血清中LH、E₂和P₄含量，说明皮下注射褪黑素能够促进卵母细胞成熟和排卵，并提高配种妊娠率。     

维生素、矿物质与能量蛋白质水平对浙东白鹅母鹅繁殖性能、血液生殖激素浓度及生殖轴相关基因mRNA相对表达量的影响

本试验通过在饲粮中添加维生素与矿物质、调整饲粮能量蛋白质水平,旨在研究其对浙东白鹅母鹅繁殖性能、血液生殖激素浓度和生殖轴相关基因mRNA相对表达量的影响.选择138只月龄相近的浙东白鹅种母鹅,按体重相近原则分为3组,分别饲喂不同的饲粮,试验期150 d,测定繁殖性能(平均产蛋数、平均蛋重、受精率和孵化率)、血液生殖激素[卵泡刺激素(FSH)、促黄体生成素(LH)、孕酮(P4)、雌二醇(E2)、催乳素(PRL)]浓度和生殖轴相关基因[促性腺激素释放激素(GnRH)、卵泡刺激素-β(FSHβ)、雌激素受体1(ESR1)、雌激素受体2(ESR2)、卵泡刺激素受体(FSHR)、催乳素(PRL)、催乳素受体(PRLR)] mRNA相对表达量的变化.结果表明:1)添加维生素与矿物质可显著提高浙东白鹅母鹅第1产蛋周期平均蛋重和受精率(P＜0.05);提高第2产蛋周期内血液FSH和P4的浓度,降低LH浓度,改变E2、P4和PRL浓度波动(P＜0.05);下调下丘脑PRLR、垂体PRL和卵巢PRLR基因的mRNA相对表达量(P＜0.05),上调卵巢ESR2基因的mRNA相对表达量(P＜0.05).2)调整饲粮能量蛋白质水平可显著提高浙东白鹅母鹅第2产蛋周期平均蛋重(P＜0.05);提高浙东白鹅第2产蛋周期内血液LH浓度,降低FSH浓度,改变E2和P4浓度波动(P＜0.05);上调下丘脑GnRH、垂体PRL和PRLR基因的mRNA相对表达量(P＜0.05),下调卵巢FSHR基因的mRNA相对表达量(P＜0.05).由此得出,添加维生素与矿物质、调整饲粮能量蛋白质水平可通过影响产蛋周期内部分血液生殖激素浓度和波动,局部调节生殖轴相关基因的mRNA相对表达量,改善浙东白鹅母鹅的繁殖性能.     

N-氨甲酰谷氨酸对奶牛超排效果及血液指标的影响

本研究旨在探讨饲喂N-氨甲酰谷氨酸（NCG）对奶牛超数排卵效果及血液生化指标的影响。选取荷斯坦育成牛16头，随机分为2组，每组8头，对照组饲喂基础饲粮，试验组在基础饲粮的基础上饲喂20 g/（d·头）的NCG，采用连续4 d递减注射FSH法测定NCG饲喂时间对供体牛超排效果的影响，分别在3次超排处理的第0、5、9天采集尾根静脉血测定激素指标及血清生化指标。结果表明，①NCG饲喂20 d时（第1次超排），试验组头均回收胚数、可用胚数、退化胚数、未受精卵数与对照组相比差异均不显著（P>0.05）；NCG饲喂50 d时（第2次超排），试验组头均回收胚数显著高于对照组（P<0.05）；NCG饲喂80 d时（第3次超排），试验组头均可用胚数显著高于对照组（P<0.05）；综合考虑3次超排效果，试验组头均回收胚数显著高于对照组（P<0.05）。②NCG饲喂时间对供体牛血清中促卵泡素、促黄体素、孕酮、雌二醇的浓度均无显著影响（P>0.05），说明NCG饲喂时间对奶牛生殖激素的分泌无影响或为次要影响因素，超排效果的变化与生殖激素变化无明显的关联作用。③NCG饲喂时间对供体牛血清中谷草转氨酶的浓度无显著影响（P>0.05）；在NCG饲喂13 d时，试验组供体牛血清中葡萄糖浓度显著高于对照组（P<0.05）；在NCG饲喂34 d时，试验组供体牛血清中尿素氮浓度显著低于对照组（P<0.05）；在NCG饲喂43和69 d时，试验组供体牛血清中一氧化氮浓度显著高于对照组（P<0.05）。综合考虑，在重复3次超排时，每头供体牛每天饲喂20 g NCG，可提高3次连续超排的回收胚数4.98枚及可用胚数1.8枚，进而降低胚胎的生产成本。     

Effect of GDF9 on Sheep Cumulus Cell Proliferation

The purpose of this study was to investigate the effect of growth differentiation factor 9 (GDF9) on the gene expression of cumulus cells expansion and hormone receptors as well as hormone secretion,in order to provide evidence for the role of GDF9 in the development of sheep cumulus cells.Sheep cumulus cells were used as the research object in this study,and were cultured for 48 h by adding different concentrations (0,50,100,200,400 ng/mL) GDF9 to low serum cell culture medium.Total RNA were extracted from the cells,using β-actin as the reference gene,Real-time quantitative PCR technology were used to detect the cumulus cells expansion related genes hyaluronic acid synthase gene 2 (HAS2),prostaglandin lead oxide synthase 2 (PTGS2),pentraxin 3 (PTX3) and hormone receptor genes follicle-stimulating hormone receptor (FSHR),luteinizing hormone receptor (LHR) and estrogen receptors (E₂R).Using the enzyme-linked immunosorbent assay (ELISA) method to test the content of E₂ and P₄.The results showed that HAS2,PTX3,FSHR,E₂R and LHR mRNA relative expression of 200 ng/mL GDF9 group was extremely significantly higher than the control group and other GDF9 groups (P<0.01),PTGS2 mRNA relative expression was extremely significantly higher than the control group and 50,400 ng/mL GDF9 groups (P<0.01),and significantly higher than 100 ng/mL GDF9 group (P<0.05).When added 400 ng/mL GDF9,the relative mRNA expression of all the mentioned-above genes were all extremely significantly lower than that of the 200 ng/mL GDF9 group.Moreover,the E₂ secretion level was extremely significantly higher than that of the control group and 50 ng/mL GDF9 group (P<0.01),significantly higher than that of the 100 ng/mL GDF9 group(P<0.05),while had no significant difference from the 200 ng/mL GDF9 group (P>0.05).When added 100,200 and 400 ng/mL GDF9,the concentration of P₄ was significantly higher than the control group (P<0.05),and there was no significant difference from the 50 ng/mL group (P>0.05),and there was no significant difference between 100,200 and 400 ng/mL GDF9 groups (P>0.05).To sum up,GDF9 could promote the expansion of sheep cumulus cells and participated in the regulation of hormone secretion of sheep cumulus cells.