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冷冻保存对山羊精子头部形态学尺寸的影响。

The effects of cryopreservation on the morphometric dimensions of caprine sperm heads.

作者信息

Gravance C G, White C, Robertson K R, Champion Z J, Casey P J

机构信息

Department of Obstetrics and Gynaecology, University of Auckland, New Zealand.

出版信息

Anim Reprod Sci. 1997 Sep;49(1):37-43. doi: 10.1016/s0378-4320(97)00053-5.

Abstract

Cryopreserved semen has been utilised in the artificial insemination of livestock species for over 40 years, even though the detrimental effects of cryopreservation on sperm function and fertility are well documented. In the present study, computer-automated sperm-head morphometry was used to determine if goat sperm-head morphometry was affected by freezing and thawing. A microscope slide was prepared from single semen samples, collected by artificial vagina, from 10 sexually active Saanen bucks. The remainder of each sample was frozen in a tris-citrate-yolk extender. After thawing, semen smears were prepared on microscope slides. All slides were stained in haematoxylin and mean sperm-head measurements of length, width, width/length, area and perimeter were determined for each slide by computer aided sperm morphometry analysis. The effects of sperm freezing on sperm-head dimensions within and among all bucks were determined. No significant (P > 0.10) freezing effect was found between fresh semen and postthaw samples for length (7.00 microns vs 7.13 microns), width (3.77 microns vs 3.87 microns), width/length (0.54 micron vs 0.54 micron), area (19.67 microns2 vs 20.57 microns2) and perimeter (18.62 microns vs 18.83 microns) when analysed across all bucks. Significant differences (P < 0.05) were however found within three bucks for area, perimeter, length and width, with the percentage increase in measurements being significantly greater than in the remaining bucks. The variability of the morphometric dimensions were not affected by freezing. The results indicate that semen freezing did not affect the overall dimensions of sperm heads across the entire population of bucks sampled. However, since sperm-head dimensions from three bucks were affected, changes in sperm-head morphometry may be indicative of spermatozoa of the semen from individuals to successfully freeze. Because the overall mean sperm-head dimensions acquired from frozen/thawed semen were not different from those of fresh semen, previously reported measurements of goat sperm heads are probably reflective of fresh semen. More importantly, retrospective studies of sperm-head morphometry and fertility may now be performed utilising extensive breeding records from frozen semen.

摘要

冷冻保存的精液已在牲畜物种的人工授精中使用了40多年,尽管冷冻保存对精子功能和生育能力的有害影响已有充分记录。在本研究中,使用计算机自动精子头部形态测量法来确定山羊精子头部形态是否受冻融影响。从10只性活跃的萨能山羊通过人工阴道采集的单个精液样本制备显微镜载玻片。每个样本的其余部分在柠檬酸三钠-卵黄稀释液中冷冻。解冻后,在显微镜载玻片上制备精液涂片。所有载玻片用苏木精染色,并通过计算机辅助精子形态分析确定每张载玻片上精子头部长度、宽度、宽长比、面积和周长的平均测量值。确定了精子冷冻对所有山羊体内和之间精子头部尺寸的影响。对所有山羊进行分析时,新鲜精液和解冻后样本在长度(7.00微米对7.13微米)、宽度(3.77微米对3.87微米)、宽长比(0.54微米对0.54微米)、面积(19.67平方微米对20.57平方微米)和周长(18.62微米对18.83微米)方面未发现显著(P>0.10)冷冻效应。然而,在三只山羊中发现面积、周长、长度和宽度存在显著差异(P<0.05),测量值的增加百分比显著高于其余山羊。形态测量尺寸的变异性不受冷冻影响。结果表明,精液冷冻并未影响所采样的整个山羊群体中精子头部的总体尺寸。然而,由于三只山羊的精子头部尺寸受到影响,精子头部形态的变化可能表明个体精液中的精子能够成功冷冻。因为从冷冻/解冻精液获得的总体平均精子头部尺寸与新鲜精液的尺寸没有差异,之前报道的山羊精子头部测量值可能反映的是新鲜精液。更重要的是,现在可以利用冷冻精液的大量繁殖记录对精子头部形态和生育能力进行回顾性研究。

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