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基于体细胞和树突膜片钳同步记录的大鼠小脑浦肯野细胞房室模型

Compartmental models of rat cerebellar Purkinje cells based on simultaneous somatic and dendritic patch-clamp recordings.

作者信息

Roth A, Häusser M

机构信息

Abteilung Zellphysiologie, Max-Planck-Institut für Medizinische Forschung, Jahnstrasse 29, 69120 Heidelberg, Germany.

出版信息

J Physiol. 2001 Sep 1;535(Pt 2):445-72. doi: 10.1111/j.1469-7793.2001.00445.x.

Abstract
  1. Simultaneous dendritic and somatic patch-clamp recordings were made from Purkinje cells in cerebellar slices from 12- to 21-day-old rats. Voltage responses to current impulses injected via either the dendritic or the somatic pipette were obtained in the presence of the selective I(h) blocker ZD 7288 and blockers of spontaneous synaptic input. Neurons were filled with biocytin for subsequent morphological reconstruction. 2. Four neurons were reconstructed and converted into detailed compartmental models. The specific membrane capacitance (C(m)), specific membrane resistance (R(m)) and intracellular resistivity (R(i)) were optimized by direct fitting of the model responses to the electrophysiological data from the same cell. Mean values were: C(m), 0.77 +/- 0.17 microF cm(-2) (mean +/- S.D.; range, 0.64-1.00 microF cm(-2)), R(m), 122 +/- 18 kOmega cm(2) (98-141 kOmega cm(2)) and R(i), 115 +/- 20 Omega cm (93-142 Omega cm). 3. The steady-state electrotonic architecture of these cells was compact under the experimental conditions used. However, somatic voltage-clamp recordings of parallel fibre and climbing fibre synaptic currents were substantially filtered and attenuated. 4. The detailed models were compared with a two-compartment model of Purkinje cells. The range of synaptic current kinetics that can be faithfully recorded using somatic voltage clamp is predicted fairly well by the two-compartment model, even though some of its underlying assumptions are violated. 5. A model of I(h) was constructed based on voltage-clamp data, and inserted into the passive compartmental models. Somatic EPSP amplitude was substantially attenuated compared to the amplitude of dendritic EPSPs at their site of generation. However, synaptic efficacy of the same quantal synaptic conductance, as measured by the somatic EPSP amplitude, was only weakly dependent on synaptic location on spiny branchlets. 6. The passive electrotonic structure of Purkinje cells is unusual in that the steady-state architecture is very compact, while voltage transients such as synaptic potentials and action potentials are heavily filtered.
摘要
  1. 对12至21日龄大鼠小脑切片中的浦肯野细胞进行了树突和胞体的同步膜片钳记录。在选择性I(h)阻滞剂ZD 7288和自发性突触输入阻滞剂存在的情况下,获得了通过树突或胞体移液管注入电流脉冲后的电压响应。神经元用生物素填充以便随后进行形态重建。2. 对四个神经元进行了重建并转化为详细的房室模型。通过将模型响应直接拟合到来自同一细胞的电生理数据,优化了比膜电容(C(m))、比膜电阻(R(m))和细胞内电阻率(R(i))。平均值为:C(m),0.77±0.17 μF cm(-2)(平均值±标准差;范围,0.64 - 1.00 μF cm(-2)),R(m),122±18 kΩ cm(2)(98 - 141 kΩ cm(2)),R(i),115±20 Ω cm(93 - 142 Ω cm)。3. 在所用实验条件下,这些细胞的稳态电紧张结构紧密。然而,平行纤维和攀缘纤维突触电流的胞体电压钳记录被大幅滤波和衰减。4. 将详细模型与浦肯野细胞的双房室模型进行了比较。尽管双房室模型的一些基本假设被违反,但它能较好地预测使用胞体电压钳可忠实记录的突触电流动力学范围。5. 基于电压钳数据构建了I(h)模型,并将其插入被动房室模型中。与树突EPSP在其产生部位的幅度相比,胞体EPSP幅度大幅衰减。然而,由胞体EPSP幅度测量的相同量子突触电导的突触效能仅微弱依赖于棘状小分支上的突触位置。6. 浦肯野细胞的被动电紧张结构不同寻常之处在于,其稳态结构非常紧密,而诸如突触电位和动作电位等电压瞬变则被大量滤波。

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