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小电导钙激活钾通道(SK)在大鼠耳蜗外毛细胞中的表达。

Expression of small-conductance calcium-activated potassium channels (SK) in outer hair cells of the rat cochlea.

作者信息

Dulon D, Luo L, Zhang C, Ryan A F

机构信息

Department of Surgery, UCSD School of Medicine, La Jolla, California 92093-0666, USA.

出版信息

Eur J Neurosci. 1998 Mar;10(3):907-15. doi: 10.1046/j.1460-9568.1998.00098.x.

Abstract

Physiological evidence suggests that SK-type Ca2+-activated K+ channels participate in ACh-induced hyperpolarization of OHCs (outer hair cells). Based on the sequences published by Kohler et al. [(1996), Science, 273: 1709), we designed degenerated primers recognizing cDNA subunits of rSK1, rSK2 and rSK3. Using this consensus set of primers, we probed by PCR a rat organ of Corti cDNA library. Two PCR products of 707 base pairs with sequence identical to rSK3 and rSK2 were obtained and cloned to generate RNA probes for in situ hybridization in the rat cochlea. The subunit rSK2 showed hybridization in the organ of Corti, at the location of the OHCs. The expression of rSK2 by OHCs was confirmed by probing with PCR a poly(A) amplified OHC cDNA library. During development, rSK2 hybridization in the organ of Corti was negative at embryonic days E16, E18 and at P0, weak at P4 and stronger from P8 to adulthood. The subunit rSK2 could also be detected in the spiral ganglion from P4 to the adult stage. Contrary to rSK2, the subunit rSK3 did not show specific hybridization in the organ of Corti at the adult stage (P120) and only a weak expression was observed at P10 and P21. Our study demonstrates expression of rSK2 in OHCs. These potassium channels are good candidates to underlie the ACh-activated K+ currents recorded during patch-clamp recordings in isolated OHCs. The expression of rSK2 in the cochlear ganglion at the adult stage suggests that SK Ca2+-activated K+ channels may also participate in the repolarization of the auditory neurons after the action potential and may influence their firing patterns.

摘要

生理学证据表明,SK型钙激活钾通道参与乙酰胆碱诱导的外毛细胞(OHCs)超极化。基于科勒等人发表的序列([(1996),《科学》,273: 1709]),我们设计了识别rSK1、rSK2和rSK3 cDNA亚基的简并引物。使用这套通用引物,我们通过聚合酶链反应(PCR)探测大鼠柯蒂氏器cDNA文库。获得了两个与rSK3和rSK2序列相同的707个碱基对的PCR产物,并进行克隆以生成用于大鼠耳蜗原位杂交的RNA探针。亚基rSK2在柯蒂氏器外毛细胞所在位置显示出杂交信号。通过用PCR探测聚腺苷酸(poly(A))扩增的外毛细胞cDNA文库,证实了外毛细胞表达rSK2。在发育过程中,柯蒂氏器中rSK2的杂交信号在胚胎第16天、第18天和出生后第0天为阴性,在出生后第4天较弱,从出生后第8天到成年期较强。亚基rSK3在成年期(出生后第120天)的柯蒂氏器中未显示特异性杂交信号,仅在出生后第10天和第21天观察到弱表达。我们的研究证明了rSK2在外毛细胞中的表达。这些钾通道很可能是在分离的外毛细胞膜片钳记录中记录到的乙酰胆碱激活钾电流的基础。成年期耳蜗神经节中rSK2的表达表明,SK钙激活钾通道也可能参与动作电位后听觉神经元的复极化,并可能影响其放电模式。

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