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大鼠海马CA3锥体细胞苔藓纤维突触处单突触兴奋性突触后电流的量子成分

Quantal components of unitary EPSCs at the mossy fibre synapse on CA3 pyramidal cells of rat hippocampus.

作者信息

Jonas P, Major G, Sakmann B

机构信息

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

出版信息

J Physiol. 1993 Dec;472:615-63. doi: 10.1113/jphysiol.1993.sp019965.

Abstract
  1. Excitatory postsynaptic currents (EPSCs) were recorded in CA3 pyramidal cells of hippocampal slices of 15- to 24-day-old rats (22 degrees C) using the whole-cell configuration of the patch clamp technique. 2. Composite EPSCs were evoked by extracellular stimulation of the mossy fibre tract. Using the selective blockers 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) and D-2-amino-5-phosphonopentanoic acid (APV), a major alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate (AMPA)/kainate receptor-mediated component and a minor NMDA receptor-mediated component with slower time course were distinguished. For the AMPA/kainate receptor-mediated component, the peak current-voltage (I-V) relation was linear, with a reversal potential close to 0 mV. The half-maximal blocking concentration of CNQX was 353 nM. 3. Unitary EPSCs of the mossy fibre terminal (MF)-CA3 pyramidal cell synapse were evoked at membrane potentials of -70 to -90 mV by low-intensity extracellular stimulation of granule cell somata using fine-tipped pipettes. The EPSC peak amplitude as a function of stimulus intensity showed all-or-none behaviour. The region of low threshold was restricted to a few micrometres. This suggests that extracellular stimulation was focal, and that the stimulus-evoked EPSCs were unitary. 4. Latency and rise time histograms of EPSCs evoked by granule cell stimulation showed narrow unimodal distributions within each experiment. The mean latency was 4.2 +/- 1.0 ms, and the mean 20-80% rise time was 0.6 +/- 0.1 ms (23 cells). When fitted within the range 0.7 ms to 20 ms after the peak, the decay of the EPSCs with the fastest rise (rise time 0.5 ms or less) could be described by a single exponential function; the mean time constant was in the range 3.0-6.6 ms with a mean of 4.8 ms (8 cells). 5. Peak amplitudes of the EPSCs evoked by suprathreshold granule cell stimulation fluctuated between trials. The apparent EPSC peak conductance in normal extracellular solution (2 mM Ca2+, 1 mM Mg2+), excluding failures, was 1 nS. Reducing the Ca2+ concentration and increasing the Mg2+ concentration reduced the mean peak amplitude in a concentration-dependent manner. 6. Peaks in EPSC peak amplitude distributions were apparent in low Ca2+ and high Mg2+. Using the criteria of equidistance and the presence of peaks and dips in the autocorrelation function, five of nine EPSC peak amplitude distributions were judged to be quantal.(ABSTRACT TRUNCATED AT 400 WORDS)
摘要
  1. 采用膜片钳技术的全细胞模式,在15至24日龄大鼠(22℃)海马脑片的CA3锥体细胞中记录兴奋性突触后电流(EPSCs)。2. 通过细胞外刺激苔藓纤维束诱发复合EPSCs。使用选择性阻断剂6-氰基-7-硝基喹喔啉-2,3-二酮(CNQX)和D-2-氨基-5-磷酸戊酸(APV),区分出一个主要的α-氨基-3-羟基-5-甲基异恶唑-4-丙酸(AMPA)/海人藻酸受体介导成分和一个时间进程较慢的次要NMDA受体介导成分。对于AMPA/海人藻酸受体介导成分,峰值电流-电压(I-V)关系呈线性,反转电位接近0 mV。CNQX的半数最大阻断浓度为353 nM。3. 使用尖端精细的移液管,通过对颗粒细胞胞体进行低强度细胞外刺激,在-70至-90 mV的膜电位下诱发苔藓纤维终末(MF)-CA3锥体细胞突触的单突触EPSCs。EPSC峰值幅度作为刺激强度的函数表现出全或无行为。低阈值区域局限于几微米。这表明细胞外刺激是局部性的,且刺激诱发的EPSCs是单突触的。4. 颗粒细胞刺激诱发的EPSCs的潜伏期和上升时间直方图在每个实验中显示出狭窄的单峰分布。平均潜伏期为4.2±1.0 ms,平均20 - 80%上升时间为0.6±0.1 ms(23个细胞)。当在峰值后0.7 ms至20 ms范围内拟合时,上升最快(上升时间0.5 ms或更短)的EPSCs的衰减可用单个指数函数描述;平均时间常数在3.0 - 6.6 ms范围内,平均值为4.8 ms(8个细胞)。5. 阈上颗粒细胞刺激诱发的EPSCs的峰值幅度在各次试验间波动。在正常细胞外溶液(2 mM Ca2 +,1 mM Mg2 +)中,排除失败情况后,表观EPSC峰值电导为1 nS。降低Ca2 +浓度并增加Mg2 +浓度以浓度依赖方式降低平均峰值幅度。6. 在低Ca2 +和高Mg2 +条件下,EPSC峰值幅度分布中出现峰值。使用等距标准以及自相关函数中峰值和谷值的存在情况,九个EPSC峰值幅度分布中有五个被判定为量子化的。(摘要截断于400字)
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d668/1160505/054c43d5324f/jphysiol00414-0645-a.jpg

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