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.     

人绒毛膜促性腺激素对母驴促排卵效果的研究

[目的]研究人绒毛膜促性腺激素（human chorionic gonadotropin,hCG）对母驴卵泡发育、排卵率、受胎率以及血清生殖激素水平的影响。[方法]选择优势卵泡直径在30~35 mm以及大于35 mm的母驴各30头,不同优势卵泡直径的母驴群体分别设置1个500 IU/头hCG处理组（n=10）、1个1 000 IU/头hCG处理组（n=10）、1个不接受hCG处理的对照组（n=10）。采用肌肉注射方法对各组母驴进行hCG处理。每隔24 h进行1次B超检查,观察各组母驴卵泡发育情况,测量卵泡直径;记录各组发生排卵的母驴数量,计算排卵率。对各组母驴进行人工输精,输精后第18天进行孕检,记录各组受胎母驴头数,计算各组受胎率。于hCG处理后0、24、48、72 h分别测定各组母驴血清中雌二醇（estradiol,E₂）和孕酮（progesterone,PROG）水平。[结果]2个群体母驴的卵泡直径随hCG注射剂量的增加而增大;优势卵泡直径大于35 mm的母驴群体中,肌肉注射hCG的2个组在处理后24 h内均出现排卵,而对照组母驴没有排卵;优势卵泡直径不同的2个母驴群体,在hCG处理48 h后排卵母驴数和排卵率与对照组相比均有所提高,其中,hCG处理后72 h,优势卵泡直径大于35 mm的母驴群体中,1 000 IU/头 hCG处理组的排卵率达到100%。2个母驴群体中,接受hCG处理的母驴,受胎率均高于对照,并且随hCG剂量的增加,受胎率有所提高;优势卵泡直径大于35 mm的母驴群体中,1 000 IU/头 hCG处理组的受胎率达到50%。2个母驴群体中,1 000 IU/头处理组在hCG处理后24 h的血清E₂浓度均较0 h时有较大幅度的提升,在0~72 h内血清PROG浓度的总体提升幅度较大。[结论]hCG处理可提高母驴的排卵率、受胎率以及血清中E₂和PROG水平,1 000 IU/头剂量的效果更好。     

青年期及产蛋期绍兴鸭和康贝尔鸭血清促性腺激素释放激素浓度的比较

采用放射免疫分析法 (RIA)测定了不同日龄 (60 ,10 0 ,2 0 0和 380日龄 )绍兴鸭和卡基 康贝尔鸭 (KhakiCampbellDuck)血清中促性腺激素释放激素 (GnRH)浓度 ,试图通过比较不同品种、性别和日龄鸭之间GnRH浓度的变化 ,探讨GnRH与产蛋性能的关系。结果表明 :绍兴鸭和康贝尔鸭母鸭血清中GnRH浓度呈现类似的年龄性变化 ,60日龄时GnRH浓度较低 ,10 0日龄时最高 ,2 0 0日龄时显著下降 ,380日龄时回升至较高水平。公鸭血清中GnRH浓度的年龄性变化与母鸭相似 ,10 0日龄时最高 ,2 0 0日龄时最低     

雌性鸭胚胎和雏鸭的抑制素和促性腺激素的分泌

测定了从孵化 ２１ ｄ 到出壳后 １ 日龄雌性鸭胚胎和初生雏 鸭外周及卵巢中具免疫活性的抑制素含量，同时也测定了促卵泡生成素（ Ｆ Ｓ Ｈ）和促黄体生成素（ Ｌ Ｈ）的含量⒚在判定有效后，用牛的抑制素放射免疫系统测定抑制素的含量⒚在第 ２１ 日至 ２５ 日时胚胎血浆中抑制素含量处于低水平， 到第 ２６ 日达到最大值，第 ２７ 日稍降后，抑制素含量在出生后快速下降⒚ Ｆ Ｓ Ｈ 含量在胚胎期无显著变化，其含量的变化与外周抑制素含量的变化相对独立⒚卵巢中抑制素含量在孵 化的第 ２５ 日有显著下降，并于第 ２６ 日时快速回升⒚卵巢和血浆中 抑制素的含量在整个研究阶段后者比前者高约 １０ 倍 ⒚血浆 Ｌ Ｈ 从第 ２１ 日开始下降，至第 ２５ 日 接着有所回升并于第 ２７ 日达第二峰值⒚总之，血浆抑制素和 Ｆ Ｓ Ｈ 在雌性鸭胚胎中不存在负反馈关系， 这预示着胚胎期抑 制素的主要作用 不是抑制垂体分 泌 Ｆ Ｓ Ｈ，同 样 Ｆ Ｓ Ｈ 也不影响抑 制素的分泌⒚     

人促性腺激素释放激素及其转运肽基因的克隆、表达与纯化

研究根据GenBank中已发表的人GnRH2基因mRNA序列以及转运肽基因核苷酸序列,借助Oli-go4.1设计了一对寡核苷酸引物,以引物3'末端的短互补序列退火形成小段双链,从而互为模板和引物,通过引导合成长达90bp的目的序列GnRH/TRS。将其克隆到pMD18-T载体上,构建重组质粒pYC1,经酶切鉴定筛选出阳性克隆pYC1,进一步的序列分析确认其中GnRH/TRS序列的正确性。pYC1经BamHI和EcoRI酶切得到GnRH/TRS片段,然后将此片段克隆到表达载体pGEX-6p-1上,构建重组质粒pYC2。经酶切鉴定筛选出阳性克隆pYC2,并转化到大肠杆菌BL21(ED3)plyS实现原核表达。IPTG诱导后成功表达出与预期大小相符的约28ku的融合蛋白,光密度扫描对表达产物进行初步定量,表达产物约占菌体总蛋白的33.3%,表达产物经GlutathioneSepharose4B层析纯化后,得到了纯度较高的GST-GnRH/TRS融合蛋白,为进一步科研和实际应用奠定基础。     

泌乳奶山羊非繁殖季节诱发发情研究

奶山羊具有明显的季节性发情的特点,在非繁殖季节里,泌乳母羊体内的催乳素水平要高于繁殖季节,从而抑制了卵巢的生理机能。本研究采用具有抑制催乳素分泌作用的溴隐亭(CB-154,Parlodel)药物来降低母羊体内的催乳素水平,配合使用促性腺激素,提高泌乳母羊的诱发发情率,使配种后不返情率达到80％(28/35)。     

不同剂量激素处理对小鼠超数排卵及产仔性能的影响

研究不同激素组合处理对小鼠超数排卵及产仔性能的影响,以便获得适宜的超数排卵的激素使用剂量。分别用不同剂量孕马血清促性腺激素(PMSG)和人绒毛膜促性腺激素(HCG)注入小鼠体内进行超数排卵处理,比较激素处理后小鼠卵母细胞成熟率、产仔数、胎儿出生重及性别比。结果显示,5 IU PMSG+5 IU HCG剂量组和10 IU PMSG+10 IU HCG剂量组合相比较,卵母细胞成熟率差异不显著(P0.05),5 IU PMSG+5 IU HCG剂量组平均产仔数最高;10 IU PMSG+10 IU HCG剂量组的胎儿平均初生重最高,但产仔数少。综合以上,在实验中,5 IU PMSG+5 IU HCG剂量组合进行超排处理效果最好。     

Effects of melatonin,p-chlorophenylalanine,and α-methylparatyrosine on plasma gonadotropin level and ovarian activity in the catfish,Heteropneustes fossilis: A study correlating changes in hypothalamic monoamines

The effects of (ip, 10 injections over 20 days) of melatonin (75 g 100 g^–1 BW), the serotonin (5-HT)-synthesis blocker, para-cholorophenylalanine (p-CPA, 10 mg 100g^{–1 BW}) and the catecholamine-synthesis blocker, -methylparatyrosine (-MPT, 10 mg 100 g^–1 BW) on gonadotropin (GTH) secretion and ovarian activity were studied in ^{Heteropneustes fossilis} during late preparatory to early prespawning (April–May). The treatments resulted in significant reductions of plasma GTH and estradiol-17 levels, the gonadosomatic index, frequency distribution of vitellogenic and postvitellogenic oocytes, and ovarian and serum ³²p-labelled alkali-labile phosphoprotein (a marker of vitellogenic activity). Most of the oocytes were nonvitellogenic or had undergone atretic changes. The hepatic ³²-phosphoprotein content increased significantly over the saline control value. The effects were similar and pronounced in the p-CPA and melatonin-treated groups but were moderate in the -MPT-treated group. Hypothalamic 5-HT content and turnover were significantly inhibited in the p-CPA and melatonin-treated groups but the content and turnover of catecholamines were not. The -MPT treatment decreased significantly the content and turnover of dopamine (DA), noradrenaline (NA), and adrenaline (A) but did not influence the 5-HT content or turnover. These results suggest that 5-HT, NA and A are stimulatory to GTH secretion and that melatonin may act on the serotonergic system to inhibit the pituitary-gonadal axis.to whom correspondence should be addressed.A part of the results was presented at the International Workshop on Pineal gland: Its molecular signals and published as an abstract in Neuroendocrinol. Lett. 14: 399 pp., 1992.