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鸡胚成骨细胞培养物细胞中钙激活钾通道的鉴定

Identification of Ca(2+)-activated K+ channels in cells of embryonic chick osteoblast cultures.

作者信息

Ravesloot J H, van Houten R J, Ypey D L, Nijweide P J

机构信息

Department of Physiology and Physiological Physics, Leiden University, The Netherlands.

出版信息

J Bone Miner Res. 1990 Dec;5(12):1201-10. doi: 10.1002/jbmr.5650051203.

Abstract

Primary cultures of embryonic chick osteoblasts consist of a heterogeneous cell population. Patch clamp measurements were done on 1- to 5-day-old osteoblasts, osteocytes, fibroblastlike cells, and cells that could not be classified on morphologic criteria. The measurements showed the omnipresence of depolarization-activated high-conductance channels in cell-attached patches. The whole-cell experiments showed an outward rectifying conductance activating at positive membrane potentials. Channels underlying the latter conductance were found to be K+ conducting in outside-out membrane patches. The activation potential of the outward rectifying K+ conductance shifted to negative membrane potentials upon increasing the intracellular Ca2+ concentration within the range of 10(-8)-10(-3.2) M. The same happened with the activation potential of the K+ channels found in outside-out patches. Finally, inside-out patch experiments directly demonstrated the dependency of the activation potential of K+ channels on Ca2+ ions. Thus the identity and main characteristics of Ca2(+)-activated K+ channels expressed by the various cell types present in chick osteoblast cultures have now been established. Decreased input resistances were found in cells of cultures more than 2 days old. This is consistent with the establishment of electrical coupling between the cells. Functions in which Ca2(+)-activated K+ channels could play a role are discussed.

摘要

鸡胚成骨细胞的原代培养物由异质细胞群体组成。对1至5日龄的成骨细胞、骨细胞、成纤维样细胞以及无法根据形态学标准分类的细胞进行了膜片钳测量。测量结果显示,在细胞贴附膜片中,去极化激活的高电导通道普遍存在。全细胞实验表明,在正膜电位下激活了外向整流电导。在外侧向外膜片中发现,构成后者电导的通道是钾离子传导性的。当细胞内钙离子浓度在10(-8)-10(-3.2)M范围内增加时,外向整流钾电导的激活电位向负膜电位移动。在外侧向外膜片中发现的钾通道的激活电位也出现同样情况。最后,内侧向外膜片实验直接证明了钾通道激活电位对钙离子的依赖性。因此,现已确定鸡胚成骨细胞培养物中存在的各种细胞类型所表达的钙激活钾通道的特性和主要特征。在培养超过2天的细胞中发现输入电阻降低。这与细胞间电偶联的建立是一致的。文中还讨论了钙激活钾通道可能发挥作用的功能。

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