Lapointe S, Ahmad I, Buhr M M, Lambert R D, Sirard M A
Department des Sciences Animales, Université Laval, Ste-Foy, QC, Canada.
J Dairy Sci. 1996 Dec;79(12):2155-62. doi: 10.3168/jds.S0022-0302(96)76591-8.
Because the different portions of the female genital tract act in many ways on sperm metabolism, the current study was undertaken to modulate the survival and fertilizing ability of bovine semen by incorporation of products from the oviduct or the follicle in extenders before freezing. Motility rates at 6 h in vitro showed a net positive effect when biological factors from total retentate or from a fraction of bovine follicular fluid (total retentate = 43%; fraction 2 = 54%), oviductal cell culture (total retentate = 43%; fraction 2 = 58%), or granulosa cell culture (total retentate = 43%; fraction 3 = 53%) were added to the extenders compared with the addition of BSA (31%). Fraction 3 of granulosa cell culture retentate also had a significant stimulatory effect on the number of sperm that penetrated mucus of cows in estrous compared with BSA (n = 205 vs. n = 159). The intracellular sperm Ca2+ concentrations were very different across treatments after thawing. Sperm from straws with BSA had the highest concentration. At 4 h, intracellular Ca2+ concentration increased for all treatments, except that for sperm treated with BSA and Ca alone, internal Ca2+ declined. Heparin plus Ca stimulated a greater internalization of Ca2+ than did Ca alone for retentate from bovine follicular fluid, oviductal cell culture, and BSA treatments: glucose consistently and significantly reduced internalization. In vitro fertilization rates were similar, and no significant differences were observed across treatments.
由于雌性生殖道的不同部分在许多方面对精子代谢产生作用,因此开展了本研究,通过在冷冻前将来自输卵管或卵泡的产物添加到稀释液中,来调节牛精液的存活和受精能力。与添加牛血清白蛋白(BSA,31%)相比,当将来自总截留物或牛卵泡液的一部分(总截留物 = 43%;组分2 = 54%)、输卵管细胞培养物(总截留物 = 43%;组分2 = 58%)或颗粒细胞培养物(总截留物 = 43%;组分3 = 53%)的生物因子添加到稀释液中时,体外6小时的活力显示出净正向效应。与BSA相比,颗粒细胞培养截留物的组分3对发情期母牛精子穿透黏液的数量也有显著的刺激作用(n = 205 vs. n = 159)。解冻后不同处理间精子细胞内钙离子浓度差异很大。含有BSA的细管中的精子浓度最高。在4小时时,除了单独用BSA和钙离子处理的精子内部钙离子浓度下降外,所有处理的细胞内钙离子浓度均升高。对于来自牛卵泡液、输卵管细胞培养物和BSA处理的截留物,肝素加钙离子比单独钙离子刺激更多的钙离子内流:葡萄糖持续且显著降低钙离子内流。体外受精率相似,各处理间未观察到显著差异。