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兔近端曲小管的跨上皮电阻和细胞膜电阻

Transepithelial and cell membrane electrical resistances of the rabbit proximal convoluted tubule.

作者信息

Lapointe J Y, Laprade R, Cardinal J

出版信息

Am J Physiol. 1984 Oct;247(4 Pt 2):F637-49. doi: 10.1152/ajprenal.1984.247.4.F637.

Abstract

A technique using double-barreled perfusion pipettes and intracellular microelectrodes was developed to measure transepithelial, apical, and basolateral membrane electrical resistances in isolated rabbit proximal convoluted tubules (PCT). This technique has been tested successfully with respect to cable analysis: the transepithelial resistance (RT) did not change with tubule length and the measured core resistance of the lumen (RC) varied according to prediction with lumen diameter and perfusate resistivity. In control solutions, a linear I-V relationship was observed at the entry of the tubule for current varying from -300 to +300 nA. The mean RT was 1,050 +/- 70 omega X cm (n = 33) (a specific resistance of 8.2 omega X cm2). Bath proteins and large variations in transtubular hydrostatic pressure had no significant effect on RT, whereas RT was not systematically related to transepithelial PD or to the sodium-to-chloride permeability ratio (n = 22). Perfusate substitution of 50 mM NaCl by mannitol increased RT by 21% (n = 7) but the same maneuver in the peritubular solution had no significant effect after a 5-min equilibration period. The ratio of apical to basolateral cell membrane resistance (RA/RBL) determined with intracellular microelectrodes was 3.1 +/- 0.3 (n = 27) in control solutions and increased within 1 min by 36% (n = 8) when glucose and alanine were replaced by mannitol in the perfusate solution. Using simultaneous initial changes in transepithelial and basolateral potential differences when glucose and alanine were removed, the individual values of RA and RBL were determined. Mean RBL was 4,900 +/- 990 omega X cm (39 +/- 1.3 omega X cm2) and mean RA was 15,000 +/- 4,300 omega X cm (118 +/- 33 omega X cm2).

摘要

开发了一种使用双管灌注移液管和细胞内微电极的技术,以测量离体兔近端曲管(PCT)中的跨上皮、顶端和基底外侧膜电阻。该技术已通过电缆分析成功测试:跨上皮电阻(RT)不随小管长度变化,并且测量的管腔核心电阻(RC)根据管腔直径和灌注液电阻率的预测而变化。在对照溶液中,在小管入口处观察到电流在-300至+300 nA之间变化时呈线性I-V关系。平均RT为1,050±70Ω×cm(n = 33)(比电阻为8.2Ω×cm2)。浴蛋白和跨管静水压力的大变化对RT没有显著影响,而RT与跨上皮PD或钠氯渗透率比没有系统关联(n = 22)。用甘露醇替代50 mM NaCl的灌注液使RT增加21%(n = 7),但在5分钟平衡期后,在肾小管周围溶液中进行相同操作没有显著影响。在对照溶液中,用细胞内微电极测定的顶端与基底外侧细胞膜电阻比(RA/RBL)为3.1±0.3(n = 27),当灌注液中葡萄糖和丙氨酸被甘露醇替代时,在1分钟内增加36%(n = 8)。利用去除葡萄糖和丙氨酸时跨上皮和基底外侧电位差的同时初始变化,确定了RA和RBL的个体值。平均RBL为4,900±990Ω×cm(39±1.3Ω×cm2),平均RA为15,000±4,300Ω×cm(118±33Ω×cm2)。

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