Burt J M, Spray D C
Department of Physiology, University of Arizona, Tucson 85724.
Proc Natl Acad Sci U S A. 1988 May;85(10):3431-4. doi: 10.1073/pnas.85.10.3431.
The activity of gap junction channels between pairs of neonatal rat heart cells in culture was studied under control conditions and during uncoupling procedures by using dual whole-cell voltage clamp techniques. Under control conditions gap junctional conductance ranged from 0.05 to 35 nS. In cell pairs exhibiting low gap junctional conductance (less than 500 pS), single-channel events with a unitary conductance of 53 +/- 2 pS (5 experiments; 186 events) were apparent. Event duration and open-time probability were estimated to be 0.95 sec and 0.17, respectively. When the junctional conductance in well-coupled cell pairs (with initial junctional conductance, greater than 5 nS) was reduced by cytoplasmic acidification or application of heptanol, single-channel events could be visualized. Compared to low-conductance controls, unitary channel conductance was unaltered (for acidification the conductance was 58 +/- 3 pS in 11 experiments with 253 events; for heptanol the conductance was 61 +/- 1 pS in 2 experiments with 171 events), while the probability of channels being open was decreased. The constancy of unitary channel conductance under control conditions and during uncoupling procedures suggests that opening and closing of the gap junction channel are all-or-none processes during which no stable subconductance states are formed.
运用双全细胞膜片钳技术,在对照条件下以及解偶联过程中,对培养的新生大鼠心肌细胞对之间缝隙连接通道的活性进行了研究。在对照条件下,缝隙连接电导范围为0.05至35 nS。在缝隙连接电导较低(小于500 pS)的细胞对中,可明显观察到单通道事件,其单通道电导为53±2 pS(5次实验,186个事件)。事件持续时间和开放时间概率分别估计为0.95秒和0.17。当通过细胞质酸化或应用庚醇降低紧密偶联细胞对(初始缝隙连接电导大于5 nS)中的连接电导时,可观察到单通道事件。与低电导对照组相比,单通道电导未改变(对于酸化,在11次实验中的253个事件中,电导为58±3 pS;对于庚醇,在2次实验中的171个事件中,电导为61±1 pS),而通道开放的概率降低。在对照条件下和解偶联过程中,单通道电导的恒定表明,缝隙连接通道的开放和关闭是全或无的过程,在此过程中不会形成稳定的亚电导状态。