利用分离的奶牛X精子冷冻精液生产体内性控胚胎试验

Production of Sexing Embryos Using X-Sorted Sperm Holstein Dairy Cow in Vivo

采用常规精液和分离的X精子冷冻精液对115头超数排卵的青年和经产荷斯坦母牛人工输精生产体内性控胚胎。结果表明,利用2支和3支性控精液给超排青年母牛输精,头均分别获得6.8枚±4.8枚和6.3枚±3.2枚可用胚胎,与常规精液的超排效果(7.2枚±5.2枚)无明显差异;利用2支和3支性控精液给超排经产母牛输精,头均可用胚胎分别为7.5枚±4.5枚和8.6枚±5.6枚,与常规精液的超排效果(6.1枚±3.2枚)相比,分别增加23%(P>0.05)和41%(P>0.05),无显著差异,但解冻后性控精子会明显影响经产母牛的受精率;对用性控精液输精生产的12枚抽样胚胎采用LAMP法进行性别鉴定并移植,结果获得了91.7%的雌性率和36.4%的受胎率。结果提示,X精子的分离过程会在一定程度上影响受精率,用分离的X精子冷冻精液对超排母牛进行人工授精生产体内性控胚胎是可行的。

One hundred and eleven ovulated cows and Holstein heifers were inseminated with X-sorted and X-unsorted sperm to product sexing embryos in vivo. The control group inseminated with the unsorted frozen-thawed X-sorted sperm 12h afer the onset of estrus in superovluated dairy cow. Group Ⅰ and Group Ⅱ were inseminated with two units（2 million sperm per unit） and three units, respectviely. Results were as follows.Numbers of available embryos per flush in Group I （6.8＋4.8）and Group Ⅱ （6.3±3.2） in superovluated Holstein heifers were similar with the control Group（7.2± 5.2）. There were no significant differences among them; Comapred with the control group（6.1±3.2）, the number of available embryos per flush in Group Ⅰ （7.5±4. 5）and Group Ⅱ （8.6 ± 5.6） in superovluated Holstein cows increased by 23 M （ P 〉 0.05） and 41Z （ P 〉0.05）, respectively. However, the fertilization rate of cows inseminated with thawed X-sorted sperm was significant lower than that of cows inseminated with unsorted sperm（ P 〈0.05）;12 embryos samples derived from the X-sorted sperm were sexed by LAMP method and transferred to the recipients, and obtained 91.7M female rate and 36.4% conception rate. The results indicated that the sorting process may have some damage in the sperm that may compromise fertilization. According to the available embryos, in vivo production of sexing embryos using X-sorted sperm in superovluated Holstein dairy cow is feasible.