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来自大鼠海马切片CA1和CA3锥体细胞的分离膜片中的谷氨酸受体通道。

Glutamate receptor channels in isolated patches from CA1 and CA3 pyramidal cells of rat hippocampal slices.

作者信息

Jonas P, Sakmann B

机构信息

Max-Planck-Institut für medizinische Forschung, Abteilung Zellphysiologie, Heidelberg, Germany.

出版信息

J Physiol. 1992 Sep;455:143-71. doi: 10.1113/jphysiol.1992.sp019294.

Abstract
  1. Currents activated by glutamate receptor (GluR) agonists were recorded from outside-out patches isolated from the soma of visually identified pyramidal neurones of the CA3 and CA1 region of rat hippocampal slices. alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA), L-glutamate (L-Glu), and kainate (KA) were delivered either by bath application through perfusion of the recording chamber or by rapid application via a piezo-driven two-barrelled fast application system. 2. Bath application of each of the three agonists activated inward currents in all patches (n = 134) at holding potentials of -50 or -60 mV. The current amplitude increased in size between 3 to 30 microM-AMPA and 100 microM to 1 mM-KA. With this slow mode of bath application, the responses showed no apparent desensitization even at saturating concentrations of AMPA (30 microM) and KA (1 mM). 3. The ratio of currents activated by 30 microM-AMPA and 300 microM-KA showed a characteristic difference between CA3 and CA1 neurones. The ratio was 0.242 +/- 0.028 (mean +/- S.E.M., n = 16) for CA3 cell patches and 0.097 +/- 0.012 (n = 8) for CA1 cell patches indicating that GluRs in the two cell populations are different. 4. The steady-state current-voltage relations (I-Vs) for AMPA- and KA-activated currents showed pronounced outward rectification for both cell types (when the main cations are Na+ in the bath and Cs+ in the pipette solution). The current reversed close to 0 mV and the ratio of chord conductances 80 mV on either side of the reversal potential was 2.66 for KA-activated currents in CA3 cell patches and 2.60 in CA1 cell patches. AMPA-activated currents showed a time-dependent increase after steps to positive membrane potentials and a decrease after steps to negative voltages, indicating that a gating process is responsible for outward rectification of the steady-state I-V. 5. The permeability (P) of GluR channels was high for Na+ as compared to Cs+ for both cell types (PNa/PCs = 0.88 and 0.84). The permeability was low for N-methyl-D-glucamine+ (PNMG/PCs < or = 0.03) and Ca2+ (PCa/PCs < or = 0.05). 6. The current noise level increased during application of AMPA or KA. Apparent single-channel conductances obtained from fluctuation analysis were higher for AMPA than for KA, but similar for both cell types. In CA3 cell patches, AMPA activated channels with an apparent chord conductance of 7.2 pS, KA of 3.0 pS conductance.(ABSTRACT TRUNCATED AT 400 WORDS)
摘要
  1. 从大鼠海马切片CA3和CA1区经视觉识别的锥体神经元胞体分离的外向膜片上记录谷氨酸受体(GluR)激动剂激活的电流。α-氨基-3-羟基-5-甲基-4-异恶唑丙酸(AMPA)、L-谷氨酸(L-Glu)和海人酸(KA)通过灌流记录室进行浴式给药,或通过压电驱动的双管快速给药系统进行快速给药。2. 三种激动剂中的每一种进行浴式给药时,在-50或-60 mV的钳制电位下,所有膜片(n = 134)均激活内向电流。电流幅度在3至30 μM-AMPA和100 μM至1 mM-KA之间增大。采用这种缓慢的浴式给药方式,即使在AMPA(30 μM)和KA(1 mM)的饱和浓度下,反应也未显示出明显的脱敏现象。3. 30 μM-AMPA和300 μM-KA激活的电流比值在CA3和CA1神经元之间表现出特征性差异。CA3细胞片的比值为0.242±0.028(平均值±标准误,n = 16),CA1细胞片的比值为0.097±0.012(n = 8),表明这两种细胞群体中的GluRs不同。4. AMPA和KA激活电流的稳态电流-电压关系(I-Vs)在两种细胞类型中均表现出明显的外向整流(当浴液中的主要阳离子为Na+且移液管溶液中的主要阳离子为Cs+时)。电流在接近0 mV时反转,CA3细胞片中KA激活电流在反转电位两侧80 mV处的弦电导比值为2.66,CA1细胞片中为2.60。AMPA激活的电流在膜电位正向阶跃后呈时间依赖性增加,在负向阶跃后减小,表明门控过程是稳态I-V外向整流的原因。5. 两种细胞类型中,GluR通道对Na+的通透性高于Cs+(PNa/PCs = 0.88和0.84)。对N-甲基-D-葡糖胺+(PNMG/PCs≤0.03)和Ca2+(PCa/PCs≤0.05)的通透性较低。6. 在应用AMPA或KA期间,电流噪声水平增加。通过波动分析获得的表观单通道电导对于AMPA高于KA,但在两种细胞类型中相似。在CA3细胞片中,AMPA激活的通道表观弦电导为7.2 pS,KA激活的通道电导为3.0 pS。(摘要截断于400字)
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/829c/1175637/a5ac8b173dae/jphysiol00427-0152-a.jpg

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