Yazejian B, Byerly L
Department of Biological Sciences, University of Southern California, Los Angeles 90089-0371.
J Membr Biol. 1989 Jan;107(1):63-75. doi: 10.1007/BF01871084.
Isolated nerve cells from Lymnaea stagnalis were studied using the internal-perfusion and patch-clamp techniques. Patch excision frequently activated a voltage-independent Ba2+-permeable channel with a slope conductance of 27 pS at negative potentials (50 mM Ba2+). This channel is not seen in patches on healthy cells and, unlike the voltage-dependent Ca channel, is not labile in isolated patches. The activity of the channel in inside-out patches is unaffected by intracellular ATP, Ca2+ below 1 mM or the catalytic subunit of cAMP-dependent protein kinase but is reversibly blocked by millimolar intracellular Ca2+ or Ba2+. The channel can be activated in on-cell patches by either internal perfusion with high Ca2+ or the long-term internal perfusion of low Ca2+ solutions not containing ATP. These channels may carry the inward Ca2+ current which causes a regenerative increase in intracellular Ca+ when snail neurons are perfused with high Ca2+ solutions. High internal Ca2+, or long periods of internal perfusion with ATP-free solutions, induces an increase in a resting (-50 mV) whole-cell Ba2+ conductance. This conductance can be turned off by returning the intracellular perfusate to a low Ca2+ solution containing ATP and Mg2+. The activity of this channel appears to have an opposite dependence on intracellular conditions to that of the voltage-dependent Ca channel.
利用内部灌注和膜片钳技术研究了椎实螺分离的神经细胞。膜片切除经常激活一种电压非依赖性的Ba2+通透通道,在负电位(50 mM Ba2+)下其斜率电导为27 pS。这种通道在健康细胞的膜片中看不到,并且与电压依赖性Ca通道不同,在分离的膜片中不稳定。外向膜片通道的活性不受细胞内ATP、低于1 mM的Ca2+或cAMP依赖性蛋白激酶催化亚基的影响,但被毫摩尔浓度的细胞内Ca2+或Ba2+可逆性阻断。通过用高Ca2+进行内部灌注或长期用不含ATP的低Ca2+溶液进行内部灌注,可在细胞贴附膜片中激活这些通道。当蜗牛神经元用高Ca2+溶液灌注时,这些通道可能携带内向Ca2+电流,导致细胞内Ca+的再生性增加。高细胞内Ca2+或长时间用无ATP溶液进行内部灌注会导致静息(-50 mV)全细胞Ba2+电导增加。通过将细胞内灌注液恢复为含有ATP和Mg2+的低Ca2+溶液,这种电导可以关闭。该通道的活性似乎对细胞内条件的依赖性与电压依赖性Ca通道相反。