Wheeler M B, Seidel G E
Animal Reproduction Laboratory, Colorado State University, Fort Collins.
Gamete Res. 1989 Feb;22(2):193-204. doi: 10.1002/mrd.1120220207.
Bovine spermatozoa were incubated in vitro with lysophosphatidylserine (LPS), lysophosphatidylcholine (LPC), lysophosphatidylethanolamine (LPE), lysophosphatidylinositol (LPI), or trypsin. Capacitation of sperm was evaluated by penetration of the zonae pellucidae of dead bovine oocytes. Capacitation times could be shortened to 3 h or less by treatment of spermatozoa with each of these lysophospholipids (LPLs) (P less than .05). The maximum oocyte penetration percentages for individual LPLs were 40% for 10 microM LPS. 24% for 160 microM LPC, 31% for 320 microM LPE, and 19% for 320 microM LPI. Capacitation also was facilitated (P less than .01) by trypsin treatment of spermatozoa. Spermatozoa treated with 250 or 2,500 units/ml of trypsin penetrated more oocytes (17 and 18%) than spermatozoa treated with 0 or 25 units/ml of trypsin (0 and 3%). Spermatozoa treated with increasing concentrations of LPL showed a decrease in both the percentage of intact acrosomes and of progressively motile spermatozoa. Increasing levels of trypsin in the incubation medium also led to a decrease (P less than .05) in the percentages of intact acrosomes and a decrease (P less than .01) in the percentages of progressively motile spermatozoa. Percentages of live, ovulated oocytes fertilized by spermatozoa incubated for 1 h in LPS (86%, 6/7) were not different from those incubated for 24 h in control medium (71%, 5/7). Percentages of oocytes fertilized with both of these capacitation treatments were higher (P less than .05) than for oocytes exposed or killed or uncapacitated sperm. Rapid induction of capacitation and the acrosome reaction can be accomplished by exogenous treatment of bovine sperm with lysophospholipids or trypsin.
将牛精子与溶血磷脂酰丝氨酸(LPS)、溶血磷脂酰胆碱(LPC)、溶血磷脂酰乙醇胺(LPE)、溶血磷脂酰肌醇(LPI)或胰蛋白酶进行体外孵育。通过死牛卵母细胞透明带的穿透情况评估精子的获能。用这些溶血磷脂(LPLs)中的每一种处理精子,可将获能时间缩短至3小时或更短(P小于0.05)。单个LPLs的最大卵母细胞穿透率分别为:10微摩尔/升LPS时为40%,160微摩尔/升LPC时为24%,320微摩尔/升LPE时为31%,320微摩尔/升LPI时为19%。用胰蛋白酶处理精子也可促进获能(P小于0.01)。用250或2500单位/毫升胰蛋白酶处理的精子比用0或25单位/毫升胰蛋白酶处理的精子穿透更多的卵母细胞(分别为17%和18%,而后者为0和3%)。用浓度不断增加的LPL处理的精子,完整顶体和进行性运动精子的百分比均下降。孵育培养基中胰蛋白酶水平的升高也导致完整顶体百分比下降(P小于0.05),进行性运动精子百分比下降(P小于0.01)。在LPS中孵育1小时的精子使存活的排卵卵母细胞受精的百分比(86%,6/7)与在对照培养基中孵育24小时的百分比(71%,5/7)没有差异。这两种获能处理使卵母细胞受精的百分比均高于暴露于或杀死的或未获能精子使卵母细胞受精的百分比(P小于0.05)。通过用溶血磷脂或胰蛋白酶对外源牛精子进行处理,可快速诱导获能和顶体反应。