Zhao H B, Santos-Sacchi J
Sections of Otolaryngology and Neurobiology, Yale University School of Medicine, New Haven, Connecticut 06510, USA.
J Gen Physiol. 1998 Oct;112(4):447-55. doi: 10.1085/jgp.112.4.447.
The effects of turgor pressure-induced membrane tension on junctional coupling of Hensen cell isolates from the inner ear were evaluated by input capacitance or transjunctional conductance measurement techniques. Turgor pressure was altered by changing either pipette pressure or the osmolarities of extracellular solutions. Both positive pipette pressure and extracellular applications of hypotonic solutions, which caused cell size to concomitantly increase, uncoupled the cells as indicated by reduced input capacitance and transjunctional conductance. These changes were, in many cases, reversible and repeatable. Intracellular application of 50 microM H-7, a broad-based protein kinase inhibitor, and 10 mM BAPTA did not block the uncoupling effect of positive turgor pressure on inner ear gap junctions. The transjunctional conductance at a holding potential of -80 mV was 53.6 +/- 5.8 nS (mean +/- SEM, n = 9) and decreased approximately 40% at a turgor pressure of 1.41 +/- 0.05 kPa. Considering the coincident kinetics of cell deformation and uncoupling, we speculate that mechanical forces work directly on gap junctions of the inner ear. These results suggest that pathologies that induce imbalances in cochlear osmotic pressure regulation may compromise normal cochlear homeostasis.
通过输入电容或跨连接电导测量技术,评估了内耳亨森细胞分离物中膨压诱导的膜张力对连接耦合的影响。通过改变移液管压力或细胞外溶液的渗透压来改变膨压。正移液管压力和细胞外应用低渗溶液都会导致细胞大小同时增加,这会使细胞解偶联,表现为输入电容和跨连接电导降低。在许多情况下,这些变化是可逆且可重复的。细胞内应用50微摩尔的H-7(一种广泛的蛋白激酶抑制剂)和10毫摩尔的BAPTA并没有阻止正膨压对内耳缝隙连接的解偶联作用。在-80毫伏的保持电位下,跨连接电导为53.6±5.8纳西门子(平均值±标准误,n = 9),在1.41±0.05千帕的膨压下大约降低40%。考虑到细胞变形和解偶联的同步动力学,我们推测机械力直接作用于内耳的缝隙连接。这些结果表明,诱导耳蜗渗透压调节失衡的病理状况可能会损害正常的耳蜗内环境稳定。