Zhang T, Zhou H M, Liu Y X
State Key Laboratory of Reproductive Biology, Chinese Academy of Sciences, Beijing, China.
Mol Hum Reprod. 1997 Mar;3(3):223-31. doi: 10.1093/molehr/3.3.223.
The expression and localization of mRNA's for tissue plasminogen activator (tPA), urokinase PA (uPA), uPA receptor (uPAR) and inhibin subunits, alpha, beta A and beta B in monkey testes was investigated. Using in-situ hybridization with digoxigenin-labelled cRNA probes (dig-cRNA), we demonstrated that tPA and plasminogen activator inhibitor type 1 (PAI-1) were expressed in testes of both immature and mature rhesus monkeys. tPA mRNA was localized predominantly in Sertoli cells. Expression level was low in immature testis, increased dramatically in the adult and varied with seminiferous cycle. PAI-1 mRNA was localized mainly in germ cells except late spermatids. uPA mRNA was expressed stage-specifically in Sertoli cells of adult testis. uPA receptor mRNA was localized in germ cells of mature testis but not in spermatogonia or late spermatids. Assayed by fibrin overlay technique, PA activity in conditioned media of purified Sertoli cells (Sc) was negligible, PA activity in media obtained from co-cultured Sertoli and Leydig cells (LS), however, was significantly increased, although Leydig cells alone were not capable of producing any PA activity. Addition of follicle stimulating hormone (FSH) to the incubation medium remarkably increased PA secretion in both Sc and LS cultures. Human chronic gonadotrophin (HCG) had no significant effect on PA activity in the Sc culture but dramatically stimulated PA activity in the co-culture system. Dihydrotestosterone (DHT) did not mimic the effect of HCG. PAI-1 activity was secreted mainly by germ cells and did not differ between the two culture systems. FSH and forskolin inhibited PAI-1 secretion. Inhibin alpha, beta A and beta B subunit mRNAs were localized in Sertoli cells of adult monkey testes, with no obvious difference in the expression levels. These data suggest that PA/PAI-1 and other related factors are expressed in rhesus monkey testis under the control of various hormones, seminiferous cycle and cell-cell interactions through paracrine or autocrine regulation. Locally generated fibrinolysis may play an important role in the process of spermatogenesis.
研究了组织型纤溶酶原激活物（tPA）、尿激酶型纤溶酶原激活物（uPA）、uPA受体（uPAR）及抑制素α、βA和βB亚基的mRNA在猴睾丸中的表达及定位。使用地高辛标记的cRNA探针（地高辛-cRNA）进行原位杂交，我们证明tPA和纤溶酶原激活物抑制剂1型（PAI-1）在未成熟和成熟恒河猴的睾丸中均有表达。tPA mRNA主要定位于支持细胞。在未成熟睾丸中表达水平较低，在成年睾丸中显著增加，并随生精周期而变化。PAI-1 mRNA主要定位于除晚期精子细胞外的生殖细胞。uPA mRNA在成年睾丸的支持细胞中呈阶段特异性表达。uPA受体mRNA定位于成熟睾丸的生殖细胞，但不存在于精原细胞或晚期精子细胞中。通过纤维蛋白覆盖技术检测，纯化的支持细胞（Sc）条件培养基中的PA活性可忽略不计，然而，共培养的支持细胞和睾丸间质细胞（LS）培养基中的PA活性显著增加，尽管单独的睾丸间质细胞不能产生任何PA活性。向孵育培养基中添加促卵泡激素（FSH）可显著增加Sc和LS培养物中的PA分泌。人绒毛膜促性腺激素（HCG）对Sc培养物中的PA活性无显著影响，但在共培养系统中显著刺激PA活性。双氢睾酮（DHT）不能模拟HCG的作用。PAI-1活性主要由生殖细胞分泌，在两种培养系统中无差异。FSH和福斯可林抑制PAI-1分泌。抑制素α、βA和βB亚基mRNA定位于成年猴睾丸的支持细胞中，表达水平无明显差异。这些数据表明，PA/PAI-1和其他相关因子在恒河猴睾丸中受多种激素、生精周期及细胞间相互作用的控制，通过旁分泌或自分泌调节表达。局部产生的纤溶作用可能在精子发生过程中起重要作用。
