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用冷冻解冻的性别分选X精子或未分选精液对超排的荷斯坦-弗里生奶牛和小母牛进行授精后的胚胎生产。

Embryo production from superovulated Holstein-Friesian dairy heifers and cows after insemination with frozen-thawed sex-sorted X spermatozoa or unsorted semen.

作者信息

Peippo J, Vartia K, Kananen-Anttila K, Räty M, Korhonen K, Hurme T, Myllymäki H, Sairanen A, Mäki-Tanila A

机构信息

MTT Agrifood Research Finland, Biotechnology and Food Research, Building H, FI-31600 Jokioinen, Finland.

出版信息

Anim Reprod Sci. 2009 Mar;111(1):80-92. doi: 10.1016/j.anireprosci.2008.02.002. Epub 2008 Feb 13.

Abstract

The aim of this study was to evaluate embryo production in superovulated Holstein-Friesian dairy heifers and cows inseminated with either X-sorted spermatozoa (2 million/dose) or unsorted semen (15 million/dose). Experiment 1 at the research farm involved eight heifers, six cows and semen of one Holstein bull. All transferable embryos were diagnosed for sex. Experiment 2 included embryo collections on commercial dairy farms: X-sorted spermatozoa from three Holstein bulls were used for 59 collections on 28 farms and unsorted semen from 32 Holstein bulls were used for 179 collections on 79 farms. Superovulations were induced by eight declining doses of FSH (total of 12 ml for heifers and 19 ml for cows) starting on days 8-12 of the estrus cycle. Inseminations began 12h after the onset of estrus and were performed two to four times at 9-15 h intervals. Low-dose X-sorted inseminates were deposited into uterine horns and unsorted semen was placed into the uterine body. In Experiment 1, on average 70.3 and 75.0% of embryos recovered from heifers, and 48.4 and 100% of embryos recovered from cows were of transferable quality in X-sorted and unsorted groups, respectively. The proportion of transferable female embryos produced approximately doubled when insemination was with X-sorted spermatozoa compared to insemination with unsorted semen (heifers 96.4% versus 41.1%; cows 81.1% versus 39.8%). In Experiment 2, estimated 53.9 and 65.5% of embryos recovered from heifers, and 21.1 and 64.5% of embryos recovered from cows were of transferable quality in X-sorted and unsorted groups, respectively. Proportions of unfertilized oocytes were 21.1 and 10.6% for heifers and 56.0 and 14.4% for cows in X-sorted and unsorted groups, respectively. Consequently, cows inseminated with X-sorted spermatozoa produced significantly smaller proportions of transferable embryos (p<0.005) and significantly larger proportions of unfertilized oocytes (p<0.001) than those inseminated with unsorted semen. Proportions of quality 1 or degenerated embryos were similar for the two treatments in both heifers and cows. Within treatments, bulls did not significantly affect the proportions of transferable, unfertilized or degenerated oocytes/embryos. It was concluded that using low-dose X-sorted spermatozoa rather than normal-dose unsorted semen for the insemination of superovulated embryo donors can improve the proportion of transferable female embryos produced but this potential may not be achieved in commercial practice, particularly in cows, because of reduced fertilization rates when using low doses of X-sorted spermatozoa.

摘要

本研究的目的是评估用X染色体精子(200万/剂量)或未分选精液(1500万/剂量)对超排的荷斯坦-弗里生奶牛和小母牛授精后的胚胎生产情况。研究农场的实验1涉及8头小母牛、6头奶牛和1头荷斯坦公牛的精液。对所有可移植胚胎进行性别诊断。实验2包括在商业奶牛场收集胚胎:来自3头荷斯坦公牛的X染色体精子用于28个农场的59次收集,来自32头荷斯坦公牛的未分选精液用于79个农场的179次收集。在发情周期的第8至12天开始,用8次递减剂量的促卵泡素诱导超排(小母牛总量为12毫升,奶牛总量为19毫升)。发情开始12小时后进行授精,每隔9至15小时进行2至4次。低剂量X染色体精子授精剂注入子宫角,未分选精液注入子宫体。在实验1中,从小母牛回收的胚胎中,X染色体精子组和未分选精液组可移植质量的胚胎平均分别为70.3%和75.0%;从奶牛回收的胚胎中,这一比例分别为48.4%和100%。与未分选精液授精相比,用X染色体精子授精产生的可移植雌性胚胎比例大约翻倍(小母牛:96.4%对41.1%;奶牛:81.1%对39.8%)。在实验2中,从小母牛回收的胚胎中,X染色体精子组和未分选精液组可移植质量的胚胎估计分别为53.9%和65.5%;从奶牛回收的胚胎中,这一比例分别为21.1%和64.5%。X染色体精子组和未分选精液组中小母牛未受精卵母细胞的比例分别为21.1%和10.6%,奶牛分别为56.0%和14.4%。因此,与用未分选精液授精的奶牛相比,用X染色体精子授精的奶牛产生的可移植胚胎比例显著更低(p<0.005),未受精卵母细胞比例显著更高(p<0.001)。两种处理方式下,小母牛和奶牛中1级或退化胚胎的比例相似。在各处理组中,公牛对可移植、未受精或退化的卵母细胞/胚胎比例没有显著影响。得出的结论是,对超排胚胎供体授精时使用低剂量X染色体精子而非正常剂量未分选精液可提高产生的可移植雌性胚胎比例,但由于使用低剂量X染色体精子时受精率降低,这一潜力在商业实践中可能无法实现,尤其是在奶牛中。

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