El-Hajj Ghaoui R, Thomson P C, Leahy T, Evans G, Maxwell W M C
Centre for Advanced Technologies in Animal Genetics and Reproduction (ReproGen), Faculty of Veterinary Science, The University of Sydney, Sydney, NSW, Australia.
Reprod Domest Anim. 2007 Oct;42(5):541-9. doi: 10.1111/j.1439-0531.2006.00819.x.
Motility characteristics (assessed subjectively and with computer-assisted semen analysis) and membrane status (after staining with chlortetracycline) of washed and non-washed frozen-thawed ram spermatozoa were evaluated after incubation in buffer and buffer containing autologous whole seminal plasma or one of its two fractions: the pellet of membrane vesicles obtained by ultracentrifugation (and used at three times normal protein concentration) or the vesicle-free supernatant fraction. Whole seminal plasma and supernatant, but not membrane vesicles, improved the motility characteristics of spermatozoa after 3 and 6 h of post-thaw incubation compared with the control buffer. Resuspension and incubation with whole seminal plasma, supernatant or membrane vesicles lowered the proportion of acrosome-reacted frozen-thawed spermatozoa compared with the control buffer. Unwashed frozen-thawed semen from three rams, incubated with autologous whole seminal plasma or its fractions and inseminated using cervical or intrauterine artificial insemination, had no effect on pregnancy rates of ewes in synchronized oestrus. However, fertility was higher after laparoscopic than cervical insemination (44.9 vs 12.3%, p < 0.001). In conclusion, resuspension and incubation of frozen-thawed ram spermatozoa in autologous whole seminal plasma or its vesicle-free supernatant fraction improved their motility characteristics and, with membrane vesicles, membrane status, but these benefits were not reflected in improved fertility after cervical or intrauterine insemination.
对洗涤和未洗涤的冻融公羊精子，在缓冲液以及含有自体全精浆或其两个组分之一的缓冲液中孵育后，评估其活力特征（主观评估及计算机辅助精液分析）和膜状态（金霉素染色后）。所述精浆的两个组分分别是：通过超速离心获得的膜泡沉淀（以正常蛋白质浓度的三倍使用）或无膜泡的上清液组分。与对照缓冲液相比，全精浆和上清液（而非膜泡）在解冻后孵育3小时和6小时后改善了精子的活力特征。与对照缓冲液相比，用全精浆、上清液或膜泡重悬并孵育降低了冻融后顶体反应精子的比例。来自三只公羊的未洗涤冻融精液，与自体全精浆或其组分孵育后，采用子宫颈或子宫内人工授精，对同期发情母羊的妊娠率没有影响。然而，腹腔镜授精后的受胎率高于子宫颈授精（44.9%对12.3%，p<0.001）。总之，冻融公羊精子在自体全精浆或其无膜泡上清液组分中重悬并孵育可改善其活力特征，与膜泡一起孵育可改善膜状态，但这些益处并未在子宫颈或子宫内授精后提高受胎率方面体现出来。
