Denechaud M, Israel J M, Mishal Z, Vincent J D
Life Sci. 1987 Jun 15;40(24):2377-84. doi: 10.1016/0024-3205(87)90512-1.
Electrophysiological experiments have shown that about 50% of cultured GH3 cells (tumoral cell line, from the anterior pituitary gland) are inexcitable i.e. they do not display action potentials either spontaneously or when depolarized by a current pulse. We report here this inexcitability may be related to cellular kinetics. Thus we have studied the relationship between the various phases of the cell cycle, the electrophysiological properties of GH3/B6 cells and spontaneous or induced Prolactin and Growth Hormone (GH) release rates. Asynchronous populations of viable cells were stained with Hoechst 33 342 DNA fluorescent dye, and sorted using a flow cytometer into G1 and S phases. After selection intracellular potentials were recorded using a single glass micro-electrode; the basal or TRH stimulated rates of PRL and GH secretions were determined by RIA. Electrical properties of the cells i.e. resting potentials, input membrane resistance and excitability, reached a maximum for cells in G2+M phases. Only cells in G2+M displayed action potentials and TRH increased their secretion by 5 times for GH and by 6 times for PRL. In G1 and S phases the cells were electrically inactive and secretion rates remained at their basal levels. These findings demonstrate that the mechanism of stimulus secretion coupling is dependent upon the phases of the cell cycle.
电生理实验表明,约50%的培养GH3细胞(一种来自垂体前叶的肿瘤细胞系)是不可兴奋的,即它们既不会自发产生动作电位,也不会在电流脉冲去极化时产生动作电位。我们在此报告,这种不可兴奋性可能与细胞动力学有关。因此,我们研究了细胞周期各阶段、GH3/B6细胞的电生理特性以及自发或诱导的催乳素和生长激素(GH)释放率之间的关系。用Hoechst 33 342 DNA荧光染料对存活细胞的异步群体进行染色,并用流式细胞仪将其分选到G1期和S期。分选后,使用单根玻璃微电极记录细胞内电位;通过放射免疫分析法测定PRL和GH分泌的基础或TRH刺激率。细胞的电学特性,即静息电位、输入膜电阻和兴奋性,在G2+M期的细胞中达到最大值。只有G2+M期的细胞显示动作电位,TRH使它们的GH分泌增加5倍,PRL分泌增加6倍。在G1期和S期,细胞没有电活动,分泌率保持在基础水平。这些发现表明,刺激-分泌偶联机制取决于细胞周期的阶段。