Strege Peter R, Holm Adrian N, Rich Adam, Miller Steven M, Ou Yijun, Sarr Michael G, Farrugia Gianrico
Department of Physiology and Biophysics, Mayo Clinic and Foundation, 200 First Street SW, Rochester, MN 55905, USA.
Am J Physiol Cell Physiol. 2003 Jan;284(1):C60-6. doi: 10.1152/ajpcell.00532.2001.
A Na(+) current is present in human jejunal circular smooth muscle cells. The aim of the present study was to determine the role of the cytoskeleton in the regulation of the Na(+) current. Whole cell currents were recorded by using standard patch-clamp techniques with Cs(+) in the pipette to block K(+) currents. Cytochalasin D and gelsolin were used to disrupt the actin cytoskeleton and phalloidin to stabilize it. Colchicine was used to disassemble the microtubule cytoskeleton (and intermediate filaments) and paclitaxel to stabilize it. Acrylamide was used to disrupt the intermediate filament cytoskeleton. Perfusion of the recording chamber at 10 ml/min increased peak Na(+) current recorded from jejunal smooth muscle cells by 27 +/- 3%. Cytochalasin D and gelsolin abolished the perfusion-induced increase in Na(+) current, whereas incubation with phalloidin, colchicine, paclitaxel, or acrylamide had no effect. In conclusion, the Na(+) current expressed in human jejunal circular smooth muscle cells appears to be regulated by the cytoskeleton. An intact actin cytoskeleton is required for perfusion-induced activation of the Na(+) current.
人空肠环形平滑肌细胞中存在钠电流。本研究的目的是确定细胞骨架在钠电流调节中的作用。使用标准膜片钳技术,在微管中加入铯离子以阻断钾电流,记录全细胞电流。用细胞松弛素D和凝溶胶蛋白破坏肌动蛋白细胞骨架,用鬼笔环肽使其稳定。用秋水仙碱拆解微管细胞骨架(和中间丝),用紫杉醇使其稳定。用丙烯酰胺破坏中间丝细胞骨架。以10毫升/分钟的速度灌注记录室,可使空肠平滑肌细胞记录到的钠电流峰值增加27±3%。细胞松弛素D和凝溶胶蛋白消除了灌注诱导的钠电流增加,而用鬼笔环肽、秋水仙碱、紫杉醇或丙烯酰胺孵育则没有效果。总之,人空肠环形平滑肌细胞中表达的钠电流似乎受细胞骨架调节。灌注诱导的钠电流激活需要完整的肌动蛋白细胞骨架。