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培养的猴支持细胞中组织型纤溶酶原激活物和纤溶酶原激活物抑制剂-1的激素调节

Hormonal regulation of tissue-type plasminogen activator and plasminogen activator inhibitor type-1 in cultured monkey Sertoli cells.

作者信息

Liu Y X, Liu K, Zhou H M, Du Q, Hu Z Y, Zou R J

机构信息

State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing.

出版信息

Hum Reprod. 1995 Mar;10(3):719-27.

PMID:7540182
Abstract

Sertoli cells play a central role in the control and maintenance of spermatogenesis. Isolated Sertoli cells of mouse and rat testes have been shown to secrete plasminogen activator (PA) and a plasminogen activator inhibitor type-1 (PAI-1) in culture. In this study, we have investigated the hormonal regulation of PA and PAI-1 activities in cultured monkey Sertoli cells. Sertoli cells (5 x 10(5) cells/well) isolated from infant rhesus monkey testes were preincubated at 35 degrees C for 16 h in 24-well plates precoated with poly(D-lysine) (5 micrograms/cm2) in 0.5 ml McCoy's 5a medium containing 5% of fetal calf serum and further incubated for 48 h in 0.5 ml serum-free medium with or without various hormones or other compounds. PA as well as PAI-1 activities in the conditioned media were assayed by fibrin overlay and reverse fibrin autography techniques respectively. The Sertoli cells in vitro secreted only tissue-type PA (tPA), no detectable amount of urokinase-type PA (uPA) could be observed. Monkey Sertoli cells were also capable of secreting PAI-1. Immunocytochemical studies indicated that both tPA and PAI-1 positive staining localized in the Sertoli cells, spermatids and residual bodies of the seminiferous epithelium; Northern blot analysis further confirmed the presence of both tPA and PAI-1 mRNA in monkey Sertoli cells. Addition of follicle-stimulating hormone (FSH) or cyclic adenosine monophosphate (cAMP) derivatives or cAMP-generating agents and gonadotrophin-releasing hormone (GnRH) agonist or phorbol ester (PMA) to the cell culture significantly increased tPA activity.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

支持细胞在精子发生的调控与维持中发挥着核心作用。已证明,小鼠和大鼠睾丸的分离支持细胞在培养中会分泌纤溶酶原激活物(PA)和1型纤溶酶原激活物抑制剂(PAI - 1)。在本研究中,我们调查了培养的猴支持细胞中PA和PAI - 1活性的激素调节。从幼年恒河猴睾丸分离的支持细胞(5×10⁵个细胞/孔)在预涂有聚(D - 赖氨酸)(5微克/平方厘米)的24孔板中,于含5%胎牛血清的0.5毫升 McCoy's 5a培养基中在35℃预孵育16小时,然后在含或不含各种激素或其他化合物的0.5毫升无血清培养基中进一步孵育48小时。分别通过纤维蛋白覆盖和反向纤维蛋白自显影技术测定条件培养基中的PA以及PAI - 1活性。体外培养的支持细胞仅分泌组织型PA(tPA),未观察到可检测量的尿激酶型PA(uPA)。猴支持细胞也能够分泌PAI - 1。免疫细胞化学研究表明,tPA和PAI - 1阳性染色定位于支持细胞、生精上皮的精子细胞和残余小体中;Northern印迹分析进一步证实猴支持细胞中存在tPA和PAI - 1 mRNA。向细胞培养物中添加促卵泡激素(FSH)或环磷酸腺苷(cAMP)衍生物或cAMP生成剂以及促性腺激素释放激素(GnRH)激动剂或佛波酯(PMA)可显著增加tPA活性。(摘要截短于250字)

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