Lacerda A E, Perez-Reyes E, Wei X, Castellano A, Brown A M
Department of Molecular Physiology and Biophysics, Baylor College of Medicine, Houston, Texas 77030.
Biophys J. 1994 Jun;66(6):1833-43. doi: 10.1016/S0006-3495(94)80977-4.
We used amplifying effects of calcium channel beta subunits to identify endogenous calcium channels in Xenopus oocytes. Expression of rat brain beta 4 increased macroscopic endogenous current magnitude with a small effect on kinetics. In contrast, expression of rat brain/cardiac beta 2 produced a much larger increase in current magnitude and dramatically slowed current decay. Low concentrations of omega-conotoxin GVIA irreversibly blocked currents in both uninjected and beta 2-injected oocytes. Single channel recordings revealed both T- and N-type calcium channels with conductances of 9 and 18 pS, respectively, in uninjected oocytes and in oocytes expressing either beta subunit. Expression of either beta subunit slowed average current decay of T-type single channels. Slowing of T-type current decay by expression of beta 2 was due to reopening of the channels. N-type single channel average current decay showed little change with expression of beta 4, whereas expression of beta 2 slowed average current decay.
我们利用钙通道β亚基的放大效应来鉴定非洲爪蟾卵母细胞中的内源性钙通道。大鼠脑β4亚基的表达增加了宏观内源性电流幅度,对动力学影响较小。相比之下,大鼠脑/心脏β2亚基的表达使电流幅度有更大增加,并显著减慢了电流衰减。低浓度的ω-芋螺毒素GVIA不可逆地阻断了未注射和注射β2亚基的卵母细胞中的电流。单通道记录显示,在未注射的卵母细胞以及表达任一β亚基的卵母细胞中,均存在电导分别为9 pS和18 pS的T型和N型钙通道。任一β亚基的表达均减慢了T型单通道的平均电流衰减。β2亚基的表达导致T型电流衰减减慢是由于通道的重新开放。N型单通道平均电流衰减在β4亚基表达时变化不大,而β2亚基的表达则减慢了平均电流衰减。