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培养的软脑膜细胞间缝隙连接的特征描述

Characterization of gap junctions between cultured leptomeningeal cells.

作者信息

Spray D C, Moreno A P, Kessler J A, Dermietzel R

机构信息

Department of Neuroscience, Albert Einstein College of Medicine, Bronx, NY 10461.

出版信息

Brain Res. 1991 Dec 24;568(1-2):1-14. doi: 10.1016/0006-8993(91)91373-9.

Abstract

Leptomeningeal cells in intact meninges or dissociated and cultured for 2 h to several weeks were dye-coupled (Lucifer yellow), and voltage-clamped pairs of freshly dissociated leptomeningeal cells were well coupled electrically. Unitary conductances of junctional channels were predominantly 40-90 pS. Junctional conductance was reversibly reduced by 2 mM halothane, 1 mM heptanol and 100% CO2 and was increased by 1 mM 8 Br-cAMP. Two gap junction proteins, connexin 26 and connexin 43, were identified between leptomeningeal cells using immunocytochemical methods; Northern blot analyses of RNA isolated from cultured leptomeningeal cells showed specific hybridization to cDNAs encoding connexins 26 and 43, but not to a cDNA encoding connexin 32. These studies demonstrate co-expression of two connexins in a single cell type in the nervous system; biophysical properties do not differ significantly from those of astrocytes and cardiac myocytes, which express only connexin 43.

摘要

完整脑膜中的软脑膜细胞或解离并培养2小时至数周的软脑膜细胞进行了染料偶联(荧光黄),并且对新鲜解离的软脑膜细胞进行电压钳制时,细胞对电耦合良好。连接通道的单位电导主要为40 - 90 pS。连接电导可被2 mM氟烷、1 mM庚醇和100%二氧化碳可逆性降低,而被1 mM 8 - 溴 - cAMP增加。使用免疫细胞化学方法在软脑膜细胞之间鉴定出两种间隙连接蛋白,连接蛋白26和连接蛋白43;从培养的软脑膜细胞中分离的RNA的Northern印迹分析显示,与编码连接蛋白26和43的cDNA有特异性杂交,但与编码连接蛋白32的cDNA无杂交。这些研究证明了两种连接蛋白在神经系统的单一细胞类型中的共表达;其生物物理特性与仅表达连接蛋白43的星形胶质细胞和心肌细胞的生物物理特性没有显著差异。

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