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北美林地野牛的超数排卵与胚胎移植

Superovulation and embryo transfer in wood bison (Bison bison athabascae).

作者信息

Toosi Behzad M, Tribulo Andres, Lessard Carl, Mastromonaco Gabriela F, McCorkell Robert B, Adams Gregg P

机构信息

Department of Veterinary Biomedical Sciences, Western College of Veterinary Medicine, University of Saskatchewan, Saskatoon, Saskatchewan, Canada.

出版信息

Theriogenology. 2013 Sep 15;80(5):542-51. doi: 10.1016/j.theriogenology.2013.05.020. Epub 2013 Jul 4.

Abstract

Two experiments were done to develop an effective superovulatory treatment protocol in wood bison for the purpose of embryo collection and transfer. In experiment 1, donor bison were assigned randomly to four treatment groups (N = 5 per group) to examine the effects of method of synchronization (follicular ablation vs. estradiol-progesterone treatment) and ovarian follicular superstimulation (single slow-release vs. split dose of FSH). Recipient bison were synchronized with donor bison by either follicular ablation (N = 8) or estradiol-progesterone treatment (N = 9). In experiment 2, bison were assigned randomly to four treatment groups (N = 5 per group) to examine the ovarian response to two versus four doses of FSH, and the effect of progesterone (ovarian superstimulation with or without an intravaginal progesterone-releasing device). Donor bison were inseminated with fresh chilled wood bison semen 12 and 24 hours after treatment with GnRH (experiment 1) or LH (experiment 2). The ovarian response was assessed using ultrasonography. In experiment 1, the number of large follicles (≥ 7 mm) increased in response to both FSH treatments, but the diameter of the largest follicle detected 4 and 5 days after the start of ovarian superstimulation was greater in bison treated with a single dose of FSH than in those treated with two doses (P < 0.05). A total of 10 ova and/or embryos were collected. One blastocyst was transferred to each of five recipient bison resulting in the birth of two live wood bison calves. In experiment 2, two doses of FSH resulted in a greater number of large follicles (≥ 9 mm) on Days 4, 5, and 6 (P < 0.05) after beginning of superstimulation (Day 0), and more ovulations than four doses of FSH (11.2 ± 2.4 vs. 6.4 ± 0.8; P < 0.05). Embryo collection was performed on only five donors, and a total of 19 ova and/or embryos were recovered. In summary, fewer FSH treatments were as good or better than multiple treatments, consistent with the notion that minimizing handling stress improves the superovulatory response in bison. Follicular ablation and estradiol plus progesterone treatment were effective for inducing ovarian synchronization in embryo donor and recipient bison, and an intravaginal progesterone-releasing device during superstimulatory treatment did not influence the superovulatory response or embryo collection. Delaying ovulation-inducing treatment (GnRH or LH) to 5 days after superstimulatory treatment resulted in a greater number of ovulations and improved embryo collection efficiency (experiment 2). Embryo collection and transfer resulted in live offspring from wild wood bison.

摘要

为了进行胚胎采集和移植,开展了两项实验以制定一种有效的美洲森林野牛超数排卵处理方案。在实验1中,将供体野牛随机分为四个处理组(每组N = 5头),以研究同步化方法(卵泡消融与雌二醇 - 孕酮处理)和卵巢卵泡超刺激(单次缓释与分次剂量的促卵泡素)的效果。受体野牛通过卵泡消融(N = 8头)或雌二醇 - 孕酮处理(N = 9头)与供体野牛同步。在实验2中,将野牛随机分为四个处理组(每组N = 5头),以研究卵巢对两剂与四剂促卵泡素的反应,以及孕酮的作用(使用或不使用阴道内孕酮释放装置进行卵巢超刺激)。在使用促性腺激素释放激素(实验1)或促黄体生成素(实验2)处理后12小时和24小时,用新鲜冷藏的美洲森林野牛精液对供体野牛进行授精。使用超声检查评估卵巢反应。在实验1中,两种促卵泡素处理均使大卵泡(≥7毫米)数量增加,但在卵巢超刺激开始后4天和5天检测到的最大卵泡直径,单剂量促卵泡素处理的野牛大于两剂量处理的野牛(P < 0.05)。总共采集到10个卵子和/或胚胎。将一枚囊胚分别移植到五头受体野牛体内,产下两头美洲森林野牛活体犊牛。在实验2中,两剂促卵泡素在超刺激开始(第0天)后的第4、5和6天导致更多的大卵泡(≥9毫米)(P < 0.05),排卵数也多于四剂促卵泡素(11.2 ± 2.4对6.4 ± 0.8;P < 0.05)。仅对五头供体进行了胚胎采集,共回收19个卵子和/或胚胎。总之,较少的促卵泡素处理与多次处理效果相当或更好,这与尽量减少处理应激可改善野牛超数排卵反应的观点一致。卵泡消融和雌二醇加孕酮处理对诱导胚胎供体和受体野牛的卵巢同步化有效,超刺激处理期间使用阴道内孕酮释放装置不影响超数排卵反应或胚胎采集。将诱导排卵处理(促性腺激素释放激素或促黄体生成素)推迟至超刺激处理后5天,导致排卵数增加并提高了胚胎采集效率(实验2)。胚胎采集和移植产生了美洲森林野牛的活体后代。

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