Menconi M C, Pellegrini M, Pellegrino M
Dipartimento di Fisiologia e Biochimica G. Moruzzi, Università di Pisa, Via S. Zeno 31, 56127, Pisa, Italy.
J Membr Biol. 2001 Mar 1;180(1):65-72. doi: 10.1007/s002320010056.
The voltage dependence of stretch-activated cation channels in leech central neurons was studied in cell-free configurations of the patch-clamp technique. We established that stretch-activated channels excised from identified cell bodies of desheathed ganglia, as well as from neurons in culture, were slowly and reversibly activated by depolarizing membrane potentials. Negative pressure stimuli, applied to the patch pipette during a slow periodical modulation of membrane potential, enhanced channel activity, whereas positive pressures depressed it. Voltage-induced channel activation was observed, with soft glass pipettes, both in inside-out and outside-out membrane patches, at negative and positive reference potentials, respectively. The results presented in this study demonstrate that membrane depolarization induces slow activation of stretch-activated channels of leech central neurons. This phenomenon is similar to that found in Xenopus oocytes, however, some peculiar features of the voltage dependence in leech stretch-activated channels indicate that specific membrane-glass interactions might not necessarily be involved. Moreover, following depolarization, stretch-activated channels in membrane patches from neurons in culture exhibited significantly shorter delay to activation (sec) than their counterparts from neurons of freshly isolated ganglia (hundreds of sec).
利用膜片钳技术的无细胞模式,研究了水蛭中枢神经元中牵张激活阳离子通道的电压依赖性。我们发现,从去鞘神经节的已鉴定细胞体以及培养的神经元中分离出的牵张激活通道,会被去极化膜电位缓慢且可逆地激活。在膜电位的缓慢周期性调制期间,施加于膜片吸管的负压刺激会增强通道活性,而正压则会抑制通道活性。使用软玻璃吸管,分别在负向和正向参考电位下,在内向外和外向内膜片中均观察到了电压诱导的通道激活。本研究呈现的结果表明,膜去极化会诱导水蛭中枢神经元牵张激活通道的缓慢激活。这一现象与非洲爪蟾卵母细胞中发现的现象类似,然而,水蛭牵张激活通道电压依赖性的一些独特特征表明,特定的膜 - 玻璃相互作用不一定参与其中。此外,去极化后,培养神经元膜片中的牵张激活通道比新鲜分离神经节神经元膜片中的对应通道表现出明显更短的激活延迟(秒)(新鲜分离神经节神经元膜片中的激活延迟为数百秒)。