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牛卵泡期对促黄体生成素(LH)脉冲分泌的不同需求以及排卵前LH峰对卵母细胞受精和胚胎发育的影响

Differential requirement for pulsatile LH during the follicular phase and exposure to the preovulatory LH surge for oocyte fertilization and embryo development in cattle.

作者信息

Lindsey Bradley R, Maclellan Lisa J, Whyte Timothy R, Kinder James E, D'Occhio Michael J

机构信息

Animal Sciences and Production Group and School of Chemical and Biomedical Sciences, Central Queensland University, Rockhampton, Qld, Australia.

出版信息

Theriogenology. 2002 Dec;58(9):1651-62. doi: 10.1016/s0093-691x(02)01056-7.

Abstract

The requirement for pulsatile LH and the LH surge for the acquisition of oocyte fertilizing potential and embryo developmental competency was examined in Zebu heifers. Follicular growth was superstimulated using the GnRH agonist-LH protocol in which pulsatile LH and the preovulatory LH surge are blocked. In experiment 1, heifers were assigned on Day 7 of the estrous cycle to receive: group 1A (n = 5), 1.5 mg norgestomet (NOR) implant; group 1B (n = 5), GnRH agonist implant. Follicular growth was superstimulated with 2x daily injections of FSH from Day 10 (a.m.) to Day 13 (p.m.), with PGF2alpha injection on Day 12 (a.m.). Heifers were ovariectomized on Day 15 (a.m.) and oocytes were placed immediately into fertilization, without 24 h maturation. Respective cleavage and blastocyst development rates were: group 1A, 0/64 oocytes (0%) and 0/64 (0%); group 1B, 34/70 oocytes (48.6%) and 2/70 (2.9%). In experiment 2, heifers were assigned on Day 7 of the estrous cycle to receive: group 2A (n = 10), 1.5 mg NOR implant; group 2B (n = 10), GnRH agonist implant; group 2C (n = 10), GnRH agonist implant. Follicular growth was superstimulated as in experiment 1 above. Heifers in groups 2A and 2B received an injection of 25 mg LH on Day 14 (p.m.) and all heifers were ovariectomized on Day 15 (a.m.); oocytes were placed immediately into fertilization without 24 h maturation. Cleavage rates were similar for heifers in group 2A (84/175 oocytes, 48.0%), group 2B (61/112 oocytes, 54.5%) and group 2C (69/163, 42.3%). Blastocyst development rates were similar for heifers in group 2A (22/175 oocytes, 12.6%) and group 2B (25/112 oocytes, 22.3%) and lower (P < 0.05) for heifers in group 2C (9/163 oocytes, 5.5%). Oocytes obtained from heifers treated with GnRH agonist, without injection of exogenous LH, underwent cleavage indicating that neither pulsatile LH nor the preovulatory LH surge are obligatory for nuclear maturation in cattle oocytes. Exposure to a surge-like increase in plasma LH increased embryo developmental competency indicating that the preovulatory LH surge promotes cytoplasmic maturation. The findings have important implications for controlling the in vivo maturation of oocytes before in vitro procedures including nuclear transfer.

摘要

在瘤牛小母牛中研究了获取卵母细胞受精潜能和胚胎发育能力对促黄体生成素(LH)脉冲分泌及LH峰的需求。使用GnRH激动剂-LH方案超刺激卵泡生长,该方案会阻断LH脉冲分泌和排卵前LH峰。在实验1中,在发情周期的第7天给小母牛分配处理:1A组(n = 5),植入1.5 mg诺孕美特(NOR);1B组(n = 5),植入GnRH激动剂。从第10天上午至第13天下午每天注射2次促卵泡素(FSH)超刺激卵泡生长,在第12天上午注射前列腺素F2α(PGF2α)。在第15天上午对小母牛进行卵巢切除,卵母细胞立即用于受精,不进行24小时成熟培养。各自的卵裂率和囊胚发育率为:1A组,64个卵母细胞中0个(0%)和64个中0个(0%);1B组,70个卵母细胞中34个(48.6%)和70个中2个(2.9%)。在实验2中,在发情周期的第7天给小母牛分配处理:2A组(n = 10),植入1.5 mg NOR;2B组(n = 10),植入GnRH激动剂;2C组(n = 10),植入GnRH激动剂。如上述实验1那样超刺激卵泡生长。2A组和2B组的小母牛在第14天下午注射25 mg LH,所有小母牛在第15天上午进行卵巢切除;卵母细胞立即用于受精,不进行24小时成熟培养。2A组(175个卵母细胞中84个,48.0%)、2B组(112个卵母细胞中61个,54.5%)和2C组(163个中69个,42.3%)小母牛的卵裂率相似。2A组(175个卵母细胞中22个,12.6%)和2B组(112个卵母细胞中25个,22.3%)小母牛的囊胚发育率相似,而2C组(163个卵母细胞中9个,5.5%)小母牛的囊胚发育率较低(P < 0.05)。从用GnRH激动剂处理且未注射外源性LH的小母牛获得的卵母细胞发生了卵裂,这表明LH脉冲分泌和排卵前LH峰对于牛卵母细胞核成熟并非必需。暴露于血浆LH的类似峰样升高可提高胚胎发育能力,这表明排卵前LH峰促进细胞质成熟。这些发现对于在包括核移植在内的体外操作之前控制卵母细胞的体内成熟具有重要意义。

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