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CX43 AND CX37 GAP JUNCTION CHANNEL ACTIVITY

CX43 和 CX37 间隙连接通道活动

基本信息

批准号：
2713985
负责人：
PETER R BRINK
金额：
$ 10.9万
依托单位：
STATE UNIVERSITY NEW YORK STONY BROOK
依托单位国家：
美国
项目类别：
财政年份：
1985
资助国家：
美国
起止时间：
1985-03-01 至 1999-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/2713985
关键词：
animal tissue cyclic AMP cyclic GMP enzyme inhibitors gap junctions human tissue immunocytochemistry membrane channels membrane permeability microelectrodes protein kinase protein structure function second messengers voltage /patch clamp

项目摘要

This proposal is focused on elucidating the gating mechanisms of connexins 43 and 37 under defined ionic and expression constraints. The aims are: 1) Determine the gating characteristics of a non-transfected population of connexin43 derived gap junction channels using dual whole cell batch clamp (DWCP). Human vascular smooth muscle-cell pairs (HVSM) derived from corpora cavernosa, which are known to contain connexin43, will be used as well as NRK cell pairs which contain rat connexin43. Gap junction channel gating will be monitored by measurement of the mean open time(s), mean closed time(s), open probabilities and voltage sensitivity. Further, we ascertain whether there is cooperative gating (non-independent gating between gap junction channels. 2a) Use DWCP in N2A cells transfected with rCx43 or humanCx37 and determine the behavior of heterotypic Cx43/Cx37 gap junction channels. Homotypic behavior for Cx37 is in the literature and we present data in the preliminary results for rCx43 transfected cells. We will continue to study the homotypic forms and determine their gating characteristics for comparison with non-transfected rCx43. Homotypic refers to two identical hemichannels linked to form a gap junction channel (Cx43/Cx43) and a heterotypic refers to two non-identical hemichannels linked together (Cx43/Cx37) where each hemichannel is composed of only one connexin type. 2b) Co-transfect Cx43 and Cx37 into N2A cells (a potential model for endothelial cell intercellular communication) and monitor the multichannel behavior. The low number of channels formed in the N2A cells in combination with our analysis will reveal any gating differences between homotypic and heterotypic gap junction channels as well as gap junctions made of heteromers (heteromers=hemichannels of two different forms,Cx43 and Cx37 found in the same hemichannel). Our assumption is that heteromeric and heterotypic forms are distinguishable from each other and homotypic forms in terms of parameters such as open probabilities, mean open times, mean closed times, cooperative gating, voltage dependence and unitary conductance. 3.) Use direct patch clamp on HVSM cells and transfected N2A cells to identify gap junction channels on the surface of freshly isolated cells. 4) Elucidate gate-to-gate interactions (contingent gating) for single gap junction channels using DWCP data where large deltaVj steps are applied and modeling both macroscopic currents and unitary channel activity. We will also use osmotic loading (Zimmerberg and Parsegian, 1986) to distinguish volume changes between different conductance states of gap junction channels.

这一建议的重点是阐明连接蛋白的门控机制

项目成果

期刊论文数量（17）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Gap junctions in vascular smooth muscle.

血管平滑肌中的间隙连接。

DOI：
10.1046/j.1365-201x.1998.00439.x
发表时间：
1998
期刊：
Acta physiologica Scandinavica.
影响因子：
0
作者：
Brink,PR
通讯作者：
Brink,PR

The inhibitory chloride channel of the lobster Panulirus penicillatus neuromuscular junction.

龙虾Panulirus penicillatus神经肌肉接头的抑制性氯离子通道。

DOI：
10.1016/0742-8413(90)90018-5
发表时间：
1990
期刊：
Comparative biochemistry and physiology. C, Comparative pharmacology and toxicology
影响因子：
0
作者：
Gilat,E;Jaslove,SW
通讯作者：
Jaslove,SW

Connexin37 forms high conductance gap junction channels with subconductance state activity and selective dye and ionic permeabilities.

Connexin37 形成具有亚电导状态活性和选择性染料和离子渗透性的高电导间隙连接通道。