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小鼠培养脊髓神经元中甘氨酸和γ-氨基丁酸门控通道的阴离子渗透机制

Mechanism of anion permeation through channels gated by glycine and gamma-aminobutyric acid in mouse cultured spinal neurones.

作者信息

Bormann J, Hamill O P, Sakmann B

机构信息

Max-Planck-Institut für biophysikalische Chemie, Göttingen, F.R.G.

出版信息

J Physiol. 1987 Apr;385:243-86. doi: 10.1113/jphysiol.1987.sp016493.

DOI:10.1113/jphysiol.1987.sp016493
PMID:2443667
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1192346/
Abstract
  1. The ion-selective and ion transport properties of glycine receptor (GlyR) and gamma-aminobutyric acid receptor (GABAR) channels in the soma membrane of mouse spinal cord neurones were investigated using the whole-cell, cell-attached and outside-out patch versions of the patch-clamp technique. 2. Current-voltage (I-V) relations of transmitter-activated currents obtained from whole-cell measurements with 145 mM-Cl- intracellularly and extracellularly, showed outward rectification. In voltage-jump experiments, the instantaneous I-V relations were linear, and the steady-state I-V relations were rectifying outwardly indicating that the gating of GlyR and GABAR channels is voltage sensitive. 3. The reversal potential of whole-cell currents shifted 56 mV per tenfold change in internal Cl- activity indicating activation of Cl(-)-selective channels. The permeability ratio of K+ to Cl- (PK/PCl) was smaller than 0.05 for both channels. 4. The permeability sequence for large polyatomic anions was formate greater than bicarbonate greater than acetate greater than phosphate greater than propionate for GABAR channels; phosphate and propionate were not measurably permeant in GlyR channels. This indicates that open GlyR and GABAR channels have effective pore diameters of 5.2 and 5.6 A, respectively. The sequence of relative permeabilities for small anions was SCN- greater than I- greater than Br- greater than Cl- greater than F- for both channels. 5. GlyR and GABAR channels are multi-conductance-state channels. In cell-attached patches the single-channel slope conductances close to 0 mV membrane potential were 29, 18 and 10 pS for glycine, and 28, 17 and 10 pS for GABA-activated channels. The most frequently observed (main) conductance states were 29 and 17 pS for the GlyR and GABAR channel, respectively. 6. In outside-out patches with equal extracellular and intracellular concentrations of 145 mM-Cl-, the conductance states were 46, 30, 20 and 12 pS for GlyR channels and 44, 30, 19 and 12 pS for GABAR channels. The most frequently occurring main state was 46 pS for the GlyR and 30 pS for the GABAR channel. 7. Single-channel conductances measured in equal 140 mM concentrations of small anions on both membrane faces revealed a conductance sequence of Cl- greater than Br- greater than I- greater than SCN- greater than F- for both channels. This is nearly the inverse sequence of that found for the permeability of these ions indicating the presence of binding sites for ions in the channel.(ABSTRACT TRUNCATED AT 400 WORDS)
摘要
  1. 采用膜片钳技术的全细胞、细胞贴附式和外翻式膜片记录模式,研究了小鼠脊髓神经元胞体膜中甘氨酸受体(GlyR)和γ-氨基丁酸受体(GABAR)通道的离子选择性及离子转运特性。2. 胞内和胞外均为145 mM-Cl-时,通过全细胞记录得到的递质激活电流的电流-电压(I-V)关系呈外向整流。在电压阶跃实验中,瞬时I-V关系呈线性,稳态I-V关系呈外向整流,表明GlyR和GABAR通道的门控对电压敏感。3. 全细胞电流的反转电位随胞内Cl-活性每10倍变化而移动56 mV,表明Cl(-)选择性通道被激活。两种通道的K+与Cl-的通透率之比(PK/PCl)均小于0.05。4. 对于GABAR通道,大的多原子阴离子的通透顺序为甲酸根>碳酸氢根>乙酸根>磷酸根>丙酸根;磷酸根和丙酸根在GlyR通道中无明显通透。这表明开放的GlyR和GABAR通道的有效孔径分别为5.2 Å和5.6 Å。两种通道的小阴离子相对通透顺序均为SCN->I->Br->Cl->F-。5. GlyR和GABAR通道是多电导状态通道。在细胞贴附式膜片中,膜电位接近0 mV时,甘氨酸激活通道的单通道斜率电导为29、18和10 pS,GABA激活通道的单通道斜率电导为28、17和10 pS。GlyR和GABAR通道最常观察到的(主要)电导状态分别为29 pS和17 pS。6. 在胞外和胞内Cl-浓度均为145 mM的外翻式膜片中,GlyR通道的电导状态为46、30、20和12 pS,GABAR通道的电导状态为44、30、19和12 pS。GlyR通道最常出现的主要状态为46 pS,GABAR通道为30 pS。7. 在膜两侧小阴离子浓度均为140 mM时测量的单通道电导显示,两种通道的电导顺序均为Cl->Br->I->SCN->F-。这几乎与这些离子的通透顺序相反,表明通道中存在离子结合位点。(摘要截选至400字)