Tabares L, Ureña J, López-Barneo J
Departamento de Fisiología, Facultad de Medicina, Sevilla, Spain.
J Gen Physiol. 1989 Mar;93(3):495-519. doi: 10.1085/jgp.93.3.495.
The ionic currents of clonal Y-1 adrenocortical cells were studied using the whole-cell variant of the patch-clamp technique. These cells had two major current components: a large outward current carried by K ions, and a small inward Ca current. The Ca current depended on the activity of two populations of Ca channels, slow (SD) and fast (FD) deactivating, that could be separated by their different closing time constants (at -80 mV, SD, 3.8 ms, and FD, 0.13 ms). These two kinds of channels also differed in (a) activation threshold (SD, approximately -50 mV; FD, approximately -20 mV), (b) half-maximal activation (SD, between -15 and -10 mV; FD between +10 and +15 mV), and (c) inactivation time course (SD, fast; FD, slow). The total amplitude of the Ca current and the proportion of SD and FD channels varied from cell to cell. The amplitude of the K current was strongly dependent on the internal [Ca2+] and was almost abolished when internal [Ca2+] was less than 0.001 microM. The K current appeared to be independent, or only slightly dependent, of Ca influx. With an internal [Ca2+] of 0.1 microM, the activation threshold was -20 mV, and at +40 mV the half-time of activation was 9 ms. With 73 mM external K the closing time constant at -70 mV was approximately 3 ms. The outward current was also modulated by internal pH and Mg. At a constant pCa gamma a decrease of pH reduced the current amplitude, whereas the activation kinetics were not much altered. Removal of internal Mg produced a drastic decrease in the amplitude of the Ca-activated K current. It was also found that with internal [Ca2+] over 0.1 microM the K current underwent a time-dependent transformation characterized by a large increase in amplitude and in activation kinetics.
运用膜片钳技术的全细胞变体,对克隆的Y-1肾上腺皮质细胞的离子电流进行了研究。这些细胞有两个主要电流成分:一个由K离子携带的大的外向电流,以及一个小的内向Ca电流。Ca电流取决于两类Ca通道的活性,即缓慢(SD)失活和快速(FD)失活通道,它们可通过不同的关闭时间常数来区分(在-80 mV时,SD为3.8 ms,FD为0.13 ms)。这两类通道在以下方面也存在差异:(a)激活阈值(SD约为-50 mV;FD约为-20 mV),(b)半数最大激活(SD在-15至-10 mV之间;FD在+10至+15 mV之间),以及(c)失活时间进程(SD快;FD慢)。Ca电流的总幅度以及SD和FD通道的比例在细胞之间各不相同。K电流的幅度强烈依赖于细胞内[Ca2+],当细胞内[Ca2+]小于0.001 microM时,K电流几乎消失。K电流似乎独立于Ca内流,或仅轻微依赖于Ca内流。当细胞内[Ca2+]为0.1 microM时,激活阈值为-20 mV,在+40 mV时激活的半衰期为9 ms。在外部K为73 mM时,-70 mV时的关闭时间常数约为3 ms。外向电流也受细胞内pH和Mg的调节。在恒定的pCaγ下,pH降低会使电流幅度减小,而激活动力学变化不大。去除细胞内Mg会使Ca激活的K电流幅度急剧下降。还发现,当细胞内[Ca2+]超过0.1 microM时,K电流会经历一个时间依赖性转变,其特征是幅度和激活动力学大幅增加。