Effect of capacitation on bull sperm binding to homologous oviductal epithelium
Effect of capacitation on bull sperm binding to homologous oviductal epithelium
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DOI：
10.1095/biolreprod54.3.575
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发表时间：
1996-03-01
影响因子：
3.6
通讯作者：
Suarez, SS
中科院分区：
文献类型：
--
作者：
Lefebvre, R;Suarez, SS
精子与输卵管上皮结合被认为是调节输卵管内精子库形成的重要机制。根据在仓鼠身上的证据，我们假设获能会影响牛精子的释放，使它们能够受精。输卵管取自发情的荷斯坦小母牛的排卵侧。峡部和壶腹上皮被挤出乳汁，变成碎片，形成外翻的小泡(外植体)。将外植体放置在TALP培养基的组织培养井中，并在39℃5%CO2中孵化。冷冻解冻后的精子在滑石粉中逆流而上，在含有20mgg/ml肝素(不含葡萄糖)的滑石粉中孵育4h后获能。未获能精子稀释后立即加入获能液中。在手术后2小时内，将精子加入到外植体中并与之孵育15分钟。对精子和外植体进行录像，并对录像带进行分析，以确定每表面积结合的精子数量。方差分析显示峡部和壶腹的精子结合数/0.1mM(2)无显著差异(p>0.05)，但有获能效应(p=0.0015)。此外，金霉素标记测定的获能精子百分比，在有外植体存在的情况下保持自由游动的精子中，比在没有外植体的情况下更高(p=0.001)。总之，获能似乎与牛精子从输卵管上皮释放有关，因此可以使精子离开储存库并使卵母细胞受精。
Sperm binding to oviductal epithelium is thought to be an important mechanism regulating sperm reservoir formation in the oviduct. On the basis of evidence in the hamster, we hypothesized that capacitation affects release of bovine sperm, allowing them to fertilize. Oviducts were obtained from the ovulatory side of estrous Holstein heifers. The isthmic and ampullar epithelia were milked out and reduced to fragments, which formed everted vesicles (explants). Explants were placed in tissue culture wells in TALP medium and incubated at 39 degrees C in 5% CO2. Frozen-thawed sperm were prepared by swim-up in TALP and capacitated by incubation for 4 h in TALP with 20 mu g/ml heparin (without glucose). Uncapacitated sperm were used immediately after dilution into capacitation medium. Within 2 h of surgery, sperm were added to the explants and incubated with them for 15 min. Sperm and explants were videotaped, and the tapes were analyzed to determine the numbers of sperm bound per surface area. ANOVA did not show a difference between the number of sperm bound/0.1 mm(2) in the isthmus and ampulla (p > 0.05); however, an effect of capacitation was detected (p = 0.0015). Also, the percentage of capacitated sperm, determined by chlortetracycline labeling, was greater in sperm that remained free-swimming in the presence of explants than in the absence of explants (p = 0.001). In conclusion, capacitation appears to be involved in the release of bovine sperm from oviductal epithelium and therefore could enable sperm to leave the reservoir and fertilize oocytes.