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Intercellular Communication Between Sertoli Cell and Leydig Cell

支持细胞和间质细胞之间的细胞间通讯

基本信息

批准号：
05671329
负责人：
NOGUCHI Kazumi
金额：
$ 1.02万
依托单位：
Yokohama City University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1993
资助国家：
日本
起止时间：
1993 至 1994
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05671329/
关键词：
Sertoli Cell Leydig Cell Testosterone Paracrine Control テストマテロン

项目摘要

In the present study, we demonstrate the existence of a factor which stimulates testosterone production by mature mouse Leydig cells in a spent medium of immature rat Setoli cell cultures and show some new biochemical characteristics of this factor.Using ultrafiltration apparatus with a molecular weight cut off of 10 kDa, Sertoli cell conditioned medium SCCM was concentrated. Fifteentimes concentrated rat SCCM stimulated testosterone production in a dose dependent fashion from 25 mul up to 50 mul. The activity came to a plateau at a dose of 100 mul. Leydig cell stimulating activity was retained by both a dialysis membrane and an ultrafiltration membrane with a molecular weight cut off of 10 kDa. However, activity was reduced by heating for 30 min at 60 C and lost after incubation with 0.1% trypsin for 1 h at 37 C.The activity was not retained by a Con A-agarose column and was demonstrated only in break-through fractions. HPLC gel filtration of 15 times concentrated SCCM preparation on TSK gel G3000 SW revealed activity at about 13 kDa. Both LH and SCCM significantly increased extracellular cAMP levels suggesting the activity of adenylcyclase activation of SCCM.We checked the enzyme activities of DELTA4 pathway from pregnenolone to testosterone. With the acute stimulation of LH,no change on the series of enzyme from pregnenolone to testosterone was revealed although testosterone production was stimulated to several times higher than the control. With SCCM treatment, same results were obtained although testosterone production was increased. Our SCCM preparation failed to show the activity stimulating Leydig cell testosterone production when incubated with a maximally stimulating dose of LH (10 IU/ml).

在本研究中，我们证明了一个因素，刺激睾丸间质细胞产生睾酮的成熟小鼠睾丸间质细胞在未成熟的大鼠Setoli细胞cultures用过的medium，并显示了一些新的生化特性，这个因素，使用超滤装置与分子量截止值为10 kDa，Sertoli细胞条件培养基SCCM浓缩。从25穆尔到50穆尔，50 μ l浓缩的大鼠SCCM以剂量依赖性方式刺激睾酮产生。活性在100穆尔的剂量下达到平台。Leydig细胞刺激活性被透析膜和截留分子量为10 kDa的超滤膜保留。然而，通过在60 ℃加热30分钟，活性降低，并且在37 ℃下与0.1%胰蛋白酶孵育1小时后丧失。在TSK凝胶G3000 SW上对15倍浓缩的SCCM制剂进行HPLC凝胶过滤，显示在约13 kDa处的活性。LH和SCCM均显著增加细胞外cAMP水平，表明SCCM的腺苷酸环化酶激活活性。我们检测了从双烯醇酮到睾酮的DELTA 4途径的酶活性。在LH的急性刺激下，虽然睾酮的产生被刺激到比对照高几倍，但从双烯醇酮到睾酮的一系列酶没有显示出变化。用SCCM处理，获得相同的结果，尽管睾酮产生增加。我们的SCCM制备物在与最大刺激剂量的LH（10 IU/ml）孵育时未能显示刺激Leydig细胞睾酮产生的活性。