Ricci A J, Wu Y C, Fettiplace R
Department of Physiology, University of Wisconsin Medical School, Madison, Wisconsin 53706, USA.
J Neurosci. 1998 Oct 15;18(20):8261-77. doi: 10.1523/JNEUROSCI.18-20-08261.1998.
Mechanoelectrical transducer currents in turtle auditory hair cells adapt to maintained stimuli via a Ca2+-dependent mechanism that is sensitive to the level of internal calcium buffer. We have used the properties of transducer adaptation to compare the effects of exogenous calcium buffers in the patch electrode solution with those of the endogenous buffer assayed with perforated-patch recording. The endogenous buffer of the hair bundle was equivalent to 0.1-0.4 mM BAPTA and, in a majority of cells, supported adaptation in an external Ca2+ concentration of 70 microM similar to that in turtle endolymph. The endogenous buffer had a higher effective concentration, and the adaptation time constant was faster in cells at the high-frequency end than at the low-frequency end of the cochlea. Experiments using buffers with different Ca2+-binding rates or dissociation constants indicated that the speed of adaptation and the resting open probability of the transducer channels could be differentially regulated and imply that the endogenous buffer must be a fast, high-affinity buffer. In some hair cells, the transducer current did not decay exponentially during a sustained stimulus but displayed damped oscillations at a frequency (58-230 Hz) that depended on external Ca2+ concentration. The gradient in adaptation time constant and the tuned transducer current at physiological levels of calcium buffer and external Ca2+ suggest that transducer adaptation may contribute to hair cell frequency selectivity. The results are discussed in terms of feedback regulation of transducer channels mediated by Ca2+ binding at two intracellular sites.
海龟听觉毛细胞中的机械电换能器电流通过一种依赖于钙离子的机制适应持续刺激,该机制对内源性钙缓冲液的水平敏感。我们利用换能器适应的特性,比较了膜片钳电极溶液中外源性钙缓冲液的作用与穿孔膜片记录法测定的内源性缓冲液的作用。毛束的内源性缓冲液相当于0.1 - 0.4 mM的乙二醇双四乙酸(BAPTA),并且在大多数细胞中,在70 microM的外部钙离子浓度下支持适应,这与海龟内淋巴中的情况相似。内源性缓冲液具有更高的有效浓度,并且在耳蜗高频端的细胞中,适应时间常数比低频端的细胞更快。使用具有不同钙离子结合速率或解离常数的缓冲液进行的实验表明,适应速度和换能器通道的静息开放概率可以受到不同的调节,这意味着内源性缓冲液必须是一种快速、高亲和力的缓冲液。在一些毛细胞中,换能器电流在持续刺激期间并非呈指数衰减,而是以依赖于外部钙离子浓度的频率(58 - 230 Hz)呈现阻尼振荡。在生理水平的钙缓冲液和外部钙离子条件下,适应时间常数的梯度和调谐的换能器电流表明,换能器适应可能有助于毛细胞的频率选择性。我们从由两个细胞内位点的钙离子结合介导的换能器通道的反馈调节方面讨论了这些结果。