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野牛(Cervus elaphus songaricus)经过超数排卵后,用冷冻-解冻的性别分选 Y 精子或未分选精液人工授精后胚胎移植和性别鉴定。

Embryo transfer and sex determination following superovulated hinds inseminated with frozen-thawed sex-sorted Y sperm or unsorted semen in Wapiti (Cervus elaphus songaricus).

机构信息

College of Animal Science, Tarim University, Alar, Xinjiang, China.

出版信息

Anim Reprod Sci. 2011 Jul;126(3-4):245-50. doi: 10.1016/j.anireprosci.2011.05.006. Epub 2011 May 24.

DOI:10.1016/j.anireprosci.2011.05.006
PMID:21664079
Abstract

The aim of this study was to evaluate embryo production in superovulated wapiti hinds inseminated with either Y-sorted or unsorted semen. Eighteen hinds were allocated to three treatment groups: AI following multiple ovulation (CIDR/FSH) with 10×10(6) Y-sorted frozen-thawed semen (Y group, n=6), or 10×10(6) and 100×10(6) unsorted frozen-thawed semen for the unsorted (n=6) and the control group (n=6). The embryos from the sixth day following insemination were collected and classified. Fifteen embryos from the unsorted or the control group, and four embryos from the Y group were sex determinated based on DNA analysis of the amelogenin gene. Twenty-one embryos from the Y group and 42 embryos from the unsorted or the control group were transferred into 21 and 42 synchronized recipients via standard procedures on 6th day post estrus, respectively. There were no significant differences in the number of recovered eggs, transferable embryos, degenerated embryos or unfertilized oocytes per hind among the three groups of the control (9.2±3.6, 4.7±1.9, 3.0±2.0, 1.5±1.4), the unsorted (8.2±1.9, 4.8±0.7, 1.7±1.0, 1.7±1.0) and the Y group (8.8±4.2, 4.2±1.8, 2.2±1.2, 2.5±2.1), respectively (P>0.05). The sex ratio of embryos from the Y group (4M/0F) was significantly (P<0.05) distinct from that of the unsorted and control group (8M/7F). The sex ratio of the offspring from sexed embryos (8M/0F) was deviated significantly (P<0.05) from that of the non-sexed embryos (11M/9F). In conclusion, the results suggested that the male embryos of predicted sex can be achieved with AI of sex-sorted cryopreserved sperm. PCR amplification using the amelogenin gene primers can be applied to DNA analysis of micro samples from wapiti embryo biopsies for sex identification. The male offspring can be produced after transferred with the male embryos of predicted sex.

摘要

本研究旨在评估用 Y 分选或未分选精液对超排雌马鹿进行人工授精后的胚胎生产情况。18 头雌马鹿被分配到三个处理组:CIDR/FSH 后进行多次排卵(AI),用 10×10(6)Y 分选冷冻解冻精液(Y 组,n=6),或用 10×10(6)和 100×10(6)未分选冷冻解冻精液进行未分选(n=6)和对照组(n=6)。从授精后第六天收集胚胎并进行分类。从未分选或对照组中获得的 15 个胚胎和 Y 组中的 4 个胚胎,根据牙本质蛋白基因的 DNA 分析进行性别鉴定。21 个 Y 组的胚胎和 42 个未分选或对照组的胚胎分别通过标准程序于发情后第 6 天转移到 21 个和 42 个同步受体中。三个对照组(9.2±3.6、4.7±1.9、3.0±2.0、1.5±1.4)、未分选组(8.2±1.9、4.8±0.7、1.7±1.0、1.7±1.0)和 Y 组(8.8±4.2、4.2±1.8、2.2±1.2、2.5±2.1)之间的每头雌马鹿的回收卵数、可转移胚胎数、退化胚胎数或未受精卵数无显著差异(P>0.05)。Y 组(4M/0F)胚胎的性别比例与未分选组和对照组(8M/7F)显著不同(P<0.05)。经性别鉴定胚胎的后代性别比例(8M/0F)与未经性别鉴定胚胎的后代性别比例(11M/9F)显著偏离(P<0.05)。总之,结果表明,用预测性别的分选冷冻精液进行人工授精可以获得预期性别的雄性胚胎。使用牙本质蛋白基因引物的 PCR 扩增可应用于马鹿胚胎活检的微样本 DNA 分析,用于性别鉴定。用预测性别的雄性胚胎转移后可产生雄性后代。

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