Electrophysiological, mechanosensitive responses of Xenopus laevis oocytes to direct, isotonic increase in intracellular volume

被引:6
作者
Bryan-Sisneros, AA [1 ]
Fraser, SP [1 ]
Djamgoz, MBA [1 ]
机构
[1] Univ London Imperial Coll Sci & Technol, Dept Biol Sci, London SW7 2AZ, England
基金
英国生物技术与生命科学研究理事会;
关键词
microelectrode; injection pipette; mechanosensitive ion channel; Xenopus laevis oocyte; two-electrode voltage clamp;
D O I
10.1016/S0165-0270(03)00041-4
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
An intra-oocyte injection method for obtaining the electrophysiological response of follicle-enclosed Xenopus laevis oocytes to an increase in intracellular volume (i.e. stretch) without changing the extracellular medium is described. The response comprised a 'stretch-activated' (SA) current which was evoked by injection of an isotonic 14-70 nl droplet and had a transient, smooth profile. Ionic substitution experiments revealed that the current was carried mainly by Na+, K+ and Cl- and had a reversal potential of about -2 mV. A similar result was obtained from experiments in which the holding potential was varied between -40 and + 10 mV whilst repeatedly inducing the SA current. On average, the channel was blocked 60% by 10 muM gadolinium chloride, 58% by 50 muM amiloride, 11% by 50 muM 4,4'-diisothiocyanatostilbene-2, 2'-disulfonic acid and 63% by 50 muM 4-acetamido-4'-isothiocyanato-stilbene-2-2'-disulfonic acid. Maturation of the oocytes with 100 muM progesterone reduced the mechanosensitivity 12-fold. This injection technique is compared with other methods of eliciting mechanosensitive (MS) currents in X. laevis oocytes. These observed characteristics of the SA current are discussed in relation to the oocytes' endogenous MS cation and anion channels. (C) 2003 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:103 / 111
页数:9
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