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新鲜分离的小鼠胰腺β细胞对之间缝隙连接的生物物理特性。

Biophysical properties of gap junctions between freshly dispersed pairs of mouse pancreatic beta cells.

作者信息

Pérez-Armendariz M, Roy C, Spray D C, Bennett M V

机构信息

Department of Neuroscience, Albert Einstein College of Medicine, Bronx, New York 10461.

出版信息

Biophys J. 1991 Jan;59(1):76-92. doi: 10.1016/S0006-3495(91)82200-7.

Abstract

Coupling between beta cells through gap junctions has been postulated as a principal mechanism of electrical synchronization of glucose-induced activity throughout the islet of Langerhans. We characterized junctional conductance between isolated pairs of mouse pancreatic beta cells by whole-cell recording with two independent patch-clamp circuits. Most pairs were coupled (67%, n = 155), although the mean junctional conductance (gj) (215 +/- 110 pS) was lower than reported in other tissues. Coupling could be recorded for long periods, up to 40 min. Voltage imposed across the junctional or nonjunctional membranes had no effect on gj. Up to several hours of treatment to increase intracellular cAMP levels did not affect gj. Electrically coupled pairs did not show transfer of the dye Lucifer yellow. Octanol (2 mM) reversibly decreased gj. Lower concentrations of octanol (0.5 mM) and heptanol (0.5 mM) than required to uncouple beta cells decreased voltage-dependent K+ and Ca2+ currents in nonjunctional membranes. Although gj recorded in these experiments would be expected to be provided by current flowing through only a few channels of the unitary conductance previously reported for other gap junctions, no unitary junctional currents were observed even during reversible suppression of gj by octanol. This result suggests either that the single channel conductance of gap junction channels between beta cells is smaller than in other tissues (less than 20 pS) or that the small mean conductance is due to transitions between open and closed states that are too rapid or too slow to be resolved.

摘要

通过缝隙连接实现的β细胞间耦合,被认为是整个胰岛中葡萄糖诱导活性电同步的主要机制。我们使用两个独立的膜片钳电路,通过全细胞记录来表征分离的小鼠胰腺β细胞对之间的连接电导。大多数细胞对是耦合的(67%,n = 155),尽管平均连接电导(gj)(215±110 pS)低于其他组织中的报道。耦合可以长时间记录,长达40分钟。施加在连接膜或非连接膜上的电压对gj没有影响。长达数小时增加细胞内cAMP水平的处理,对gj没有影响。电耦合的细胞对没有显示出荧光黄染料的转移。辛醇(2 mM)可逆地降低gj。与使β细胞解偶联所需浓度相比,较低浓度的辛醇(0.5 mM)和庚醇(0.5 mM)降低了非连接膜上电压依赖性K⁺和Ca²⁺电流。尽管在这些实验中记录的gj预计仅由流经先前报道的其他缝隙连接单一电导的少数通道的电流提供,但即使在辛醇对gj进行可逆抑制期间,也未观察到单一连接电流。该结果表明,要么β细胞之间缝隙连接通道的单通道电导小于其他组织(小于20 pS),要么小的平均电导是由于开放和关闭状态之间的转换太快或太慢而无法分辨。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/baf2/1281120/ebfa111af32a/biophysj00119-0091-a.jpg

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