Albillos A, Neher E, Moser T
Department of Membrane Biophysics, Max-Planck Institute for Biophysical Chemistry, 37077 Goettingen, Germany.
J Neurosci. 2000 Nov 15;20(22):8323-30. doi: 10.1523/JNEUROSCI.20-22-08323.2000.
Patch-clamp measurements of Ca(2+) currents and membrane capacitance were performed on slices of mouse adrenal glands, using the perforated-patch configuration of the patch-clamp technique. These recording conditions are much closer to the in vivo situation than those used so far in most electrophysiological studies in adrenal chromaffin cells (isolated cells maintained in culture and whole-cell configuration). We observed profound discrepancies in the quantities of Ca(2+) channel subtypes (P-, Q-, N-, and L-type Ca(2+) channels) described for isolated mouse chromaffin cells maintained in culture. Differences with respect to previous studies may be attributable not only to culture conditions, but also to the patch-clamp configuration used. Our experiments revealed the presence of a Ca(2+) channel subtype never before described in chromaffin cells, a toxin and dihydropyridine-resistant Ca(2+) channel with fast inactivation kinetics, similar to the R-type Ca(2+) channel described in neurons. This channel contributes 22% to the total Ca(2+) current and controls 55% of the rapid secretory response evoked by short depolarizing pulses. Our results indicate that R-type Ca(2+) channels are in close proximity with the exocytotic machinery to rapidly regulate the secretory process.
采用膜片钳技术的穿孔膜片配置,对小鼠肾上腺切片进行了Ca(2+)电流和膜电容的膜片钳测量。与迄今为止大多数肾上腺嗜铬细胞电生理研究(培养的分离细胞和全细胞配置)所采用的记录条件相比,这些记录条件更接近体内情况。我们观察到,对于培养的分离小鼠嗜铬细胞,所描述的Ca(2+)通道亚型(P型、Q型、N型和L型Ca(2+)通道)数量存在显著差异。与先前研究的差异可能不仅归因于培养条件,还归因于所使用的膜片钳配置。我们的实验揭示了嗜铬细胞中一种前所未有的Ca(2+)通道亚型,一种对毒素和二氢吡啶耐药、具有快速失活动力学的Ca(2+)通道,类似于神经元中描述的R型Ca(2+)通道。该通道对总Ca(2+)电流的贡献为22%,并控制短去极化脉冲诱发的快速分泌反应的55%。我们的结果表明,R型Ca(2+)通道与胞吐机制紧密相邻,以快速调节分泌过程。