Liu Y X, Hsueh A J
Biol Reprod. 1987 May;36(4):1055-62. doi: 10.1095/biolreprod36.4.1055.
Two types of plasminogen activator (tissue-type, tPA; urokinase-type, uPA) have been demonstrated in ovarian granulosa cells, but only tPA activity was found in denuded oocytes. Immature rats were treated subcutaneously with 20 IU pregnant mare's serum gonadotropin (PMSG) to stimulate follicle maturation, followed 2 days later by an injection of 10 IU human chorionic gonadotropin (hCG) to induce ovulation. Cellular plasminogen activator activities were determined by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis followed by a fibrin-overlay technique. Cumulus-oocyte complexes from rats before and after PMSG treatment contained low amounts of tPA, but not uPA, activity. After hCG treatment, tPA activity showed a time-dependent increase, reaching a maximum at 24 h after injection. At 12 and 24 h after hCG treatment, uPA activity was also detected. The appearance of high molecular weight lysis zones further suggested the formation of plasminogen activator-inhibitor complexes. Morphological analysis indicated that the increases in oocyte tPA activity were correlated with the extent of cumulus cell expansion and dispersion. In denuded oocytes, tPA activity also progressively increased during the periovulatory period to a maximum at 24 h after hCG treatment. In contrast, neither uPA activity nor activator-inhibitor complex was detected. Secretion of the proteases was measured in the conditioned media of cumulus-oocyte complexes cultured for 24 h in vitro. Substantial increases in tPA release were found in complexes obtained at 8 and 12 h after hCG injection, with lower secretion from complexes obtained at 24 h after hCG treatment.(ABSTRACT TRUNCATED AT 250 WORDS)
在卵巢颗粒细胞中已证实存在两种纤溶酶原激活剂(组织型,tPA;尿激酶型,uPA),但在裸卵中仅发现了tPA活性。对未成熟大鼠皮下注射20国际单位孕马血清促性腺激素(PMSG)以刺激卵泡成熟,2天后注射10国际单位人绒毛膜促性腺激素(hCG)诱导排卵。通过十二烷基硫酸钠(SDS)-聚丙烯酰胺凝胶电泳,随后采用纤维蛋白覆盖技术来测定细胞纤溶酶原激活剂活性。PMSG处理前后大鼠的卵丘-卵母细胞复合体含有少量的tPA活性,但没有uPA活性。hCG处理后,tPA活性呈时间依赖性增加,在注射后24小时达到最大值。在hCG处理后12小时和24小时,也检测到了uPA活性。高分子量裂解区的出现进一步提示了纤溶酶原激活剂-抑制剂复合物的形成。形态学分析表明,卵母细胞tPA活性的增加与卵丘细胞扩展和分散的程度相关。在裸卵中,tPA活性在排卵前期也逐渐增加,在hCG处理后24小时达到最大值。相比之下,未检测到uPA活性和激活剂-抑制剂复合物。在体外培养24小时的卵丘-卵母细胞复合体的条件培养基中测量蛋白酶的分泌。在hCG注射后8小时和12小时获得的复合体中发现tPA释放大幅增加,而在hCG处理后24小时获得的复合体中分泌较低。(摘要截断于250字)