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拟南芥叶肉细胞中Ca(2+)通透、外向整流钾离子通道

Ca(2+)-permeable, outwardly-rectifying K+ channels in mesophyll cells of Arabidopsis thaliana.

作者信息

Romano L A, Miedema H, Assmann S M

机构信息

Dept. of Biology, Pennsylvania State University, University Park 16802, USA.

出版信息

Plant Cell Physiol. 1998 Nov;39(11):1133-44. doi: 10.1093/oxfordjournals.pcp.a029314.

Abstract

Arabidopsis thaliana has become a powerful tool in genetics and molecular biology. In order to use Arabidopsis as a model system for electrophysiological studies on plant cells, a detailed characterization of the transporters present in the plasma and vacuolar membranes of this species is required. We used the patch-clamp technique to study ion channels in the plasma membrane of Arabidopsis mesophyll cells. The most prominent conductance in these cells was a K(+)-selective, voltage-dependent, outwardly-rectifying channel (IK,out). In the whole-cell configuration, IK,out was observed in 100% of the cells assayed. In contrast, inward current was observed in less than 50% of the cells which were bathed in 100 mM K+, and was totally absent from cells bathed in 10 mM K+. The activation kinetics of IK,out were modulated by the external K+ concentration with a faster activation at low external K+. Tail-current analysis revealed that in addition to K+, IK,out is also permeable to Ca2+ and Ba2+. Externally applied Ba2+ also caused a voltage-dependent decrease in current magnitude, indicating that IK,out is also partially blocked by this classic K+ channel blocker. Single channels studied in outside-out patches showed Ca2+ and Ba2+ sensitivity, voltage dependence and time activation similar to that of IK,out in the whole-cell configuration. Given their permeability to Ca2+, these channels may function as an avenue for Ca2+ influx as well as K+ efflux, both of which may affect photosynthesis.

摘要

拟南芥已成为遗传学和分子生物学中的一种强大工具。为了将拟南芥用作植物细胞电生理研究的模型系统,需要对该物种质膜和液泡膜中存在的转运蛋白进行详细表征。我们使用膜片钳技术研究拟南芥叶肉细胞质膜中的离子通道。这些细胞中最显著的电导是一种K(+)选择性、电压依赖性、外向整流通道(IK,out)。在全细胞模式下,在所检测的100%的细胞中都观察到了IK,out。相比之下,在浸泡于100 mM K+中的细胞中,不到50%的细胞观察到内向电流,而浸泡于10 mM K+中的细胞则完全没有内向电流。IK,out的激活动力学受外部K+浓度调节,在低外部K+浓度下激活更快。尾电流分析表明,除了K+外,IK,out对Ca2+和Ba2+也有通透性。外部施加的Ba2+也导致电流幅度呈电压依赖性降低,表明IK,out也被这种经典K+通道阻滞剂部分阻断。在外侧向外膜片中研究的单通道显示出与全细胞模式下IK,out类似的Ca2+和Ba2+敏感性、电压依赖性和时间激活。鉴于它们对Ca2+的通透性,这些通道可能作为Ca2+内流以及K+外流的途径发挥作用,这两者都可能影响光合作用。

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