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爪蟾胚胎脊髓神经元培养物中的钙激活氯离子通道

Calcium-activated chloride channels in cultured embryonic Xenopus spinal neurons.

作者信息

Hussy N

机构信息

Department of Biology, University of California, San Diego, La Jolla 92093.

出版信息

J Neurophysiol. 1992 Dec;68(6):2042-50. doi: 10.1152/jn.1992.68.6.2042.

Abstract
  1. Single-channel currents were recorded from Xenopus spinal neurons developing in vitro using the patch-clamp technique, to identify the channels underlying the large and small macroscopic Ca(2+)-activated Cl- currents (ICl(Ca)) present in these cells. 2. Channels of large (maxi-channels; 310 pS) and smaller conductance (mini-channels; 50-60 pS) are activated by elevation of cytoplasmic Ca2+ concentration. Channel activity is not altered by subsequent removal of Ca2+ from the bath, arguing against a direct ligand-type Ca2+ dependence. The much higher incidence of channel activation in cell-attached patches from cells permeabilized with the Ca2+ ionophore A23187 than in excised patches also suggests the involvement of some unidentified intracellular factor. 3. The reversal potential of maxi-Cl- channels is not altered by changes in Na+ concentration, but is shifted in the negative direction by the substitution of Cl- by methanesulfonate on the intracellular side of the patch, indicating their anionic selectivity. 4. Maxi-Cl- channels exhibited the presence of multiple probable subconductance states and showed marked voltage-dependent inactivation above and below +/- 20 mV. 5. Examination of maxi-Cl- channels at early times in culture (6-9 h) and 24 h later did not reveal any developmental change in the characteristics described above. However, the mean open duration of the channel was found to increase twofold during this period of time. 6. The simultaneous presence of maxi- and mini-Cl- channels prevented detailed characterization of the latter. The anionic selectivity of mini-Cl- channels is suggested by their reversal potential that lies close to the Cl- equilibrium potential.(ABSTRACT TRUNCATED AT 250 WORDS)
摘要
  1. 采用膜片钳技术记录体外培养的非洲爪蟾脊髓神经元的单通道电流,以确定这些细胞中存在的大小不同的宏观钙激活氯电流(ICl(Ca))所对应的通道。2. 大电导通道(大通道;310 pS)和较小电导通道(小通道;50 - 60 pS)可被细胞质钙离子浓度升高激活。随后从浴槽中去除钙离子并不会改变通道活性,这与直接的配体型钙依赖性不符。在用钙离子载体A23187通透的细胞的细胞贴附片中通道激活的发生率远高于切除片,这也表明存在一些未明确的细胞内因子参与其中。3. 大氯通道的反转电位不会因钠离子浓度的变化而改变,但当在膜片细胞内侧用甲磺酸盐替代氯离子时,其会向负方向移动,表明其具有阴离子选择性。4. 大氯通道表现出多个可能的亚电导状态,并且在±20 mV以上和以下显示出明显的电压依赖性失活。5. 在培养早期(6 - 9小时)和24小时后对大氯通道进行检查,未发现上述特征有任何发育变化。然而,在此期间发现通道的平均开放持续时间增加了两倍。6. 大氯通道和小氯通道同时存在妨碍了对后者的详细表征。小氯通道的反转电位接近氯平衡电位,提示其具有阴离子选择性。(摘要截断于250字)

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