Department of Integrative Biology and Physiology, 621 Charles E. Young Drive S, University of California, Los Angeles, CA 90095 1606, USA.
Hear Res. 2011 Feb;272(1-2):69-84. doi: 10.1016/j.heares.2010.10.015. Epub 2010 Oct 31.
When amphibian papillar hair cells (APHCs) of the leopard frog, Rana pipiens pipiens, are osmotically challenged, they exhibit a characteristically asymmetric (rectifying) response: small decreases (5%, or less) in the extracellular solution's osmolarity do not significantly affect the cells' volume; larger decreases produce a relatively slow volume increase in APHCs, while exposure to a hyperosmotic medium leads to rapid shrinking of these cells. Furthermore, the rate of volume change appears to be a function of the rate of extracellular osmotic change. These characteristics make the application of methods devised for the estimation of the osmotic permeability coefficient (P(f)) for semipermeable membranes - i.e., those with significant permeability only to water - to APHC membrane rather futile. We have, therefore, devised a method that takes both the permeability to solutes as well as the kinetics of the osmolarity change into consideration, in order to obtain estimates of P(f) that are to a large degree independent of these factors. We have compared the new and earlier methods. Using the new method, we have estimated the P(f) of APHCs' plasma membrane to be in the 10(-2)-cm/s range, and thus significantly larger than those reported for lipid bilayers. APHC's membrane P(f) appears to be cell-size independent and insensitive to extracellular mercury. These results suggest that APHCs express water-permeable channels in their plasma membrane. Furthermore, we suggest that asymmetric and rate dependent shape changes produced by osmolarity changes in APHCs imply the presence of significant permeability to solutes. The significance of transmembrane solute transport and water channel expression in amphibian auditory hair cells is discussed.
当豹蛙(Rana pipiens pipiens)的两栖类毛细胞(APHC)受到渗透挑战时,它们会表现出一种特征性的不对称(整流)反应:细胞外液渗透压的小幅度降低(5%或更小)不会显著影响细胞体积;较大的降低会导致 APHC 相对缓慢地增加体积,而暴露于高渗介质会导致这些细胞快速收缩。此外,体积变化的速率似乎是细胞外渗透压变化速率的函数。这些特性使得应用于半透膜(即仅对水具有显著渗透性的膜)渗透压渗透性系数(P(f))估计的方法应用于 APHC 膜变得毫无意义。因此,我们设计了一种方法,该方法既考虑了溶质的渗透性,又考虑了渗透压变化的动力学,以便获得在很大程度上独立于这些因素的 P(f)估计值。我们比较了新方法和早期方法。使用新方法,我们估计 APHC 质膜的 P(f)在 10(-2)-cm/s 范围内,明显大于脂质双层报道的 P(f)。APHC 膜 P(f)似乎与细胞大小无关,并且对细胞外汞不敏感。这些结果表明 APHC 在其质膜中表达了水渗透性通道。此外,我们认为,渗透压变化引起的 APHC 不对称和速率依赖性形状变化暗示了溶质的通透性显著增加。讨论了跨膜溶质转运和水通道表达在两栖听觉毛细胞中的意义。