Yeh T H, Tsai M C, Lee S Y, Hsu M M, Tran Ba Huy P
Department of Otolaryngology, National Taiwan University Hospital, Taipei.
Hear Res. 1997 Jul;109(1-2):1-10. doi: 10.1016/s0378-5955(97)00030-0.
Ion channels on the apical membrane of epithelial cells (the surface facing the endolymph) of acutely isolated Reissner's membrane from guinea-pig cochlea were investigated by using patch-clamp technique in cell-attached and inside-out configurations. Three types of ion channel were identified: namely, a stretch-activated nonselective cation, a chloride and a potassium channel. When the pipette was filled with high-K+ endolymph-like solution, the most significant channel activity was nonselective cation channels (85/110, 77% patches). The current versus voltage relationship was linear with a unitary conductance of 22.1 +/- 0.4 pS and reversal potential (Vr) of 2.3 +/- 0.8 mV (n = 18). The channel exhibited a lower conductance (14.0 +/- 0.6 pS, n = 8) to Ca2+. The open probability was low (NPo approximately 0.1) in cell-attached configuration under +60 mV pipette potential and increased when the membrane was stretched with negative pressure. The channel was blocked by 10 microM extracellular Gd3+. The two other types of channels were a small voltage-sensitive Cl- channel (6.0 +/- 0.3 pS; 91/99, 92% patches) and a K+ channel (approximately 30 pS; 29/191, 15% patches). These channels might play roles in the regulation of cell volume, in balancing the hydrostatic pressure across Reissner's membrane and in maintaining the electrochemical composition of endolymph.
采用膜片钳技术,在细胞贴附式和内面向外式记录模式下,对豚鼠耳蜗急性分离的Reissner膜上皮细胞顶端膜(面向内淋巴的表面)上的离子通道进行了研究。鉴定出三种类型的离子通道,即牵张激活的非选择性阳离子通道、氯离子通道和钾离子通道。当移液管中充满高钾内淋巴样溶液时,最显著的通道活性是由非选择性阳离子通道产生的(110个膜片中有85个,占77%)。电流-电压关系呈线性,单通道电导为22.1±0.4 pS,反转电位(Vr)为2.3±0.8 mV(n = 18)。该通道对Ca2+的电导较低(14.0±0.6 pS,n = 8)。在细胞贴附式记录模式下,当移液管电位为+60 mV时,通道开放概率较低(NPo约为0.1),而当用负压拉伸膜片时,开放概率增加。该通道可被10 μM的细胞外Gd3+阻断。另外两种通道分别是一种小的电压敏感性Cl-通道(6.0±0.3 pS;99个膜片中有91个,占92%)和一种K+通道(约30 pS;191个膜片中有29个,占15%)。这些通道可能在调节细胞体积、平衡Reissner膜两侧的静水压力以及维持内淋巴的电化学组成方面发挥作用。