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Electrophysiological properties of testosterone-secreting cells studied with patch-clamp techniques- a comparison with neurosecretion.

用膜片钳技术研究睾酮分泌细胞的电生理特性——与神经分泌的比较。

基本信息

批准号：
61570060
负责人：
KAWA Kazuyoshi
金额：
$ 1.34万
依托单位：
Gunma 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：
1986
资助国家：
日本
起止时间：
1986 至 1987
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-61570060/
关键词：
Patch-clamp Testrosterone Calcium channels Ion channels Intercellular couplings Leydig cells 精巣

项目摘要

The testosterone-secreting cells (Leydig cells) in adult mice were isolated mechanically by perfusing intertubular spaces of testes with physiological saline. Perfusates were collected in recording chamber and the Leydig cells were identified by their morphological characteristics (diameter 13-20 <micrn>m,bright cell-border, occasionally,clustered. Indentification was confirmed reliable by the specific staining of 3<beta>-hydroxysteroid dehydrogenase.The whole-cell membranes of Leydig cells were voltage-clamped with patch-electrodes. The holding potential was -80--95 mV and the external saline contained 126 mM NaCl, 5 mM KCl and 10 mM CaCl_2. Depolarization over -40 mV evoked inward currents which decayed completely during 150 msec stimuli. The inward currents were identified as Ca^<2+>-carried current, since replacement of external Ca^<2+> with Mn^<2+> reversibly diminished the current whereas Ba^<2+> or Sr^<2+> substituted for Ca^<2+> . No measurable outward currents were evoked by the depolarization up to +40 mV. The Ca^<2+> channels may be activated by physiological changes in membrane potential, leading to an influx of Ca^<2+>.Leydig cells within a cluster possessed distinct intercellular couplings. The mean coupling ratio obtained by applying two patch electrodes to a pair of cells was 0.84. Transfer or injected dye (Lucifer Yellow) to adjacent cells was also confirmed. Intercellular coupling may paly a role in synchronizing or intensifying the endocrine activities of Leydig cells located within a cluster.

用生理盐水灌注睾丸小管间隙，机械分离成年小鼠睾丸激素分泌细胞（睾丸间质细胞）。在记录室中收集灌注液，根据其形态特征（直径13-20 < micr_bb_0 m，胞缘明亮，偶有聚集）鉴定出间质细胞。3< β >-羟基类固醇脱氢酶特异性染色证实鉴定可靠。用贴片电极对间质细胞的全细胞膜进行电压箝制。保持电位为-80—95 mV，外用盐水为126 mM NaCl、5 mM KCl和10 mM CaCl_2。-40 mV以上的去极化引起向内电流，在150毫秒的刺激期间完全衰减。向内电流被确定为Ca^<2+>携带电流，因为用Mn^<2+>取代外部Ca^<2+>可逆地减小了电流，而Ba^<2+>或Sr^<2+>取代了Ca^<2+>。除极化至+40 mV时，未产生可测量的向外电流。Ca^<2+>通道可能被膜电位的生理变化激活，导致Ca^<2+>的内流。一个簇内的间质细胞具有明显的细胞间偶联。两个贴片电极对一对细胞的平均偶联比为0.84。转移或注射染料（路西法黄）到邻近的细胞也被证实。细胞间偶联可能在同步或加强位于一个簇内的间质细胞的内分泌活动中起作用。