Ledesma A, Manes J, Cesari A, Alberio R, Hozbor F
Facultad de Ciencias Agrarias, Universidad Nacional de Mar del Plata, Balcarce, Argentina.
Reprod Domest Anim. 2014 Apr;49(2):324-32. doi: 10.1111/rda.12279. Epub 2014 Feb 3.
This study was conducted to evaluate the effect of seminal collection method (artificial vagina or electroejaculation) on the protein composition of seminal plasma and sperm quality parameters in Corriedale rams. To address this question, we assessed the effect of seminal collection method on motility, plasma membrane integrity and functionality, mitochondrial functionality and the decondensation state of nuclear chromatin in sperm cells. Volume, pH, osmolarity, protein concentration, total protein content and protein profile using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and 2-D polyacrylamide electrophoresis of seminal plasma collected with artificial vagina and electroejaculation were also analysed. The main findings from this study were that ejaculates obtained with electroejaculation had (i) a higher number of spermatozoa with intact plasma membrane and functional mitochondria and (ii) a higher proportion of seminal plasma, total protein content and relative abundance of low molecular weight proteins than ejaculates obtained with artificial vagina. Five of these proteins were identified by mass spectrometry: binder of sperm 5 precursor; RSVP14; RSVP22; epididymal secretory protein E1 and clusterin. One protein spot with molecular weight of approximately 31 kDa and isoelectric point of 4.8 was only found in the seminal plasma from electroejaculation.
本研究旨在评估采精方法（人工阴道采精或电刺激采精）对考力代公羊精浆蛋白质组成和精子质量参数的影响。为解决这个问题，我们评估了采精方法对精子活力、质膜完整性和功能、线粒体功能以及精子细胞核染色质解聚状态的影响。还分析了使用人工阴道采精和电刺激采精所获得的精浆的体积、pH值、渗透压、蛋白质浓度、总蛋白含量以及采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）和二维聚丙烯酰胺凝胶电泳的蛋白质谱。本研究的主要发现是，电刺激采精获得的射精样本具有：（i）更多质膜完整且线粒体功能正常的精子；（ii）与人工阴道采精获得的射精样本相比，精浆比例、总蛋白含量和低分子量蛋白质的相对丰度更高。通过质谱鉴定出其中5种蛋白质：精子结合蛋白5前体；RSVP14；RSVP22；附睾分泌蛋白E1和簇集蛋白。仅在电刺激采精获得的精浆中发现了一个分子量约为31 kDa、等电点为4.8的蛋白点。
