Gögelein H
Max-Planck-Institut für Biophysik, Frankfurt/Main, FRG.
Ren Physiol Biochem. 1990 Jan-Apr;13(1-2):8-25.
In the plasma membranes of mammalian proximal renal tubules single ion channels were investigated mainly in isolated tubules perfused on one side, in isolated nonperfused (collapsed) tubules and in primary cell cultures. With these techniques, the following results were obtained: in the luminal membrane of isolated one-sided perfused tubules of rabbit and mouse S3 segments, K(+)-selective channels with single-channel conductance (g) of 33 pS and 63 pS, respectively, were recorded. In primary cultures of rabbit S1 segments, a small-conductance (42 pS) as well as a large-conductance (200 pS) K+ channel were observed. The latter was Ca2(+)- and voltage-sensitive. In cultured cells a Ca2(+)-activated, nonselective cation channel with g = 25 pS was also recorded. On the other hand, an amiloride-sensitive channel with g = 12 pS, which was highly selective for Na+ over K+, was observed in the isolated perfused S3 segment. In the basolateral membrane of isolated perfused S3 segments, two types of K+ channels with g = 46 pS and 36 pS, respectively, were observed. The latter channel was not dependent on cytosolic Ca2+ in cell-excised patches. A K+ channel with g = 54 pS was recorded in isolated nonperfused S1 segments. This channel showed inward rectification and was more active at depolarizing potentials. In isolated perfused S3 segments, in addition to the K+ channels also a nonselective cation channel with g = 28 pS was observed. This channel was highly dependent on cytosolic Ca2+ in cell-free patches. It can be concluded that the K+ channels both in the luminal and contraluminal cell membrane are involved in the generation of the cell potential. Na+ channels in the luminal membrane may participate in Na+ reabsorption, whereas the function of a basolateral cation channel remains unclear. Recently, single anion-selective channels were recorded in membranes of endocytotic vesicles, isolated from rat proximal tubules. Vesicles were enlarged by the dehydration/rehydration method and investigated with the patch clamp technique. The Cl- channel had a conductance of 73 pS, the current-voltage curve was linear and the channel inactivated at high negative clamp potentials. It is suggested that this channel is responsible for charge neutrality during active H+ uptake into the endosomes.
在哺乳动物近端肾小管的质膜中,主要在一侧灌注的分离肾小管、分离的非灌注(塌陷)肾小管和原代细胞培养物中研究了单离子通道。通过这些技术,获得了以下结果:在兔和小鼠S3段一侧灌注的分离肾小管的管腔膜中,分别记录到单通道电导(g)为33 pS和63 pS的K⁺选择性通道。在兔S1段的原代培养物中,观察到一个小电导(42 pS)以及一个大电导(200 pS)的K⁺通道。后者对Ca²⁺和电压敏感。在培养细胞中还记录到一个g = 25 pS的Ca²⁺激活的非选择性阳离子通道。另一方面,在分离的灌注S3段中观察到一个g = 12 pS的氨氯地平敏感通道,它对Na⁺的选择性远高于K⁺。在分离的灌注S3段的基底外侧膜中,分别观察到两种g = 46 pS和36 pS的K⁺通道。后者通道在细胞内翻片时不依赖于胞质Ca²⁺。在分离的非灌注S1段中记录到一个g = 54 pS的K⁺通道。该通道表现出内向整流,在去极化电位时更活跃。在分离的灌注S3段中,除了K⁺通道外,还观察到一个g = 28 pS的非选择性阳离子通道。该通道在无细胞片时高度依赖于胞质Ca²⁺。可以得出结论,管腔和对侧细胞膜中的K⁺通道都参与了细胞电位的产生。管腔膜中的Na⁺通道可能参与Na⁺重吸收,而基底外侧阳离子通道的功能尚不清楚。最近,在从大鼠近端肾小管分离的内吞小泡膜中记录到了单阴离子选择性通道。通过脱水/再水化方法使小泡膨胀,并用膜片钳技术进行研究。Cl⁻通道的电导为73 pS,电流-电压曲线呈线性,且通道在高负钳制电位下失活。有人认为该通道负责在内体主动摄取H⁺过程中的电荷中和。