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自体和同源精浆对大量稀释的公猪精子活力的比较影响。

Comparative effects of autologous and homologous seminal plasma on the viability of largely extended boar spermatozoa.

作者信息

Caballero I, Vazquez J M, Centurión F, Rodríguez-Martinez H, Parrilla I, Roca J, Cuello C, Martinez E A

机构信息

Department of Medicine and Surgery (Animal Reproduction), Faculty of Veterinary Medicine, University of Murcia, Murcia, Spain.

出版信息

Reprod Domest Anim. 2004 Oct;39(5):370-5. doi: 10.1111/j.1439-0531.2004.00530.x.

Abstract

Sperm handling, associated to artificial reproduction technologies (ART) such as in vitro fertilization (IVF) or the use of flow cytometry for cell analysis or sorting imposes volumetric extension of the sperm suspension and decreases sperm viability, presumably because of the removal of seminal plasma (SP) components. This study evaluated whether a 10% v/v of autologous SP (retrieved from the same donor boar) or homologous SP (e.g. from any of the four fertile boars included, other than the one providing the spermatozoa) would differently affect the viability of boar spermatozoa subjected to large extension in a simple saline medium [phosphate-buffered saline and 0.1% ethylenediaminetetraacetic acid (EDTA), PBSm] to a concentration of 0.3 x 10(6) spermatozoa/ml and incubated for 2 h at 30 degrees C. Sperm viability was monitored as membrane integrity [using the fluorophore carboxyfluorescein diacetate (C-FDA) and propidium iodide (PI)], mitochondrial function (using the fluorophore R-123) and motility characteristics [using Computer Assisted Sperm Analysis (CASA)]. Substraction of the SP and extension followed by incubation in PBSm significantly (p < 0.05) decreased sperm viability, which could be restored by addition of autologous SP. Furthermore, exposure of the extended spermatozoa to homologous SP (from any other individual boar) significantly (p < 0.05) varied with the source of the sire; some boars exerting beneficial effects (even surpassing the effects of the autologous SP; p < 0.05) while at least one boar negatively (p < 0.05) influencing the viability of the incubated spermatozoa. It is concluded that SP should be present when incubating highly extended spermatozoa. As a result of the obvious differences among boars, it would be advantageous to examine the ability of SP to maintain sperm viability prior to the use of SP pools during sperm handling in vitro.

摘要

精子处理与体外受精(IVF)等人工生殖技术(ART)相关,或者用于细胞分析或分选的流式细胞术,这会导致精子悬浮液的体积扩大,并降低精子活力,推测这是由于精浆(SP)成分的去除。本研究评估了10%(v/v)的自体SP(从同一供体公猪获取)或同源SP(例如从包括的四只可育公猪中的任何一只获取,但不是提供精子的那只公猪)是否会对在简单盐溶液培养基[磷酸盐缓冲盐水和0.1%乙二胺四乙酸(EDTA),PBSm]中大量稀释至浓度为0.3×10⁶精子/ml并在30℃孵育2小时的公猪精子活力产生不同影响。精子活力通过膜完整性[使用荧光团羧基荧光素二乙酸酯(C-FDA)和碘化丙啶(PI)]、线粒体功能(使用荧光团R-123)和运动特征[使用计算机辅助精子分析(CASA)]进行监测。去除SP并稀释后在PBSm中孵育显著(p<0.05)降低了精子活力,添加自体SP可恢复活力。此外,稀释后的精子暴露于同源SP(来自任何其他个体公猪)时,其活力显著(p<0.05)因父本来源而异;一些公猪产生有益影响(甚至超过自体SP的影响;p<0.05),而至少有一只公猪对孵育精子的活力产生负面影响(p<0.05)。得出的结论是,在孵育高度稀释的精子时应存在SP。由于公猪之间存在明显差异,在体外精子处理过程中使用SP池之前,检测SP维持精子活力的能力将是有利的。

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