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用全细胞记录法测量完整两栖动物视网膜神经元的被动膜参数。

Measurement of passive membrane parameters with whole-cell recording from neurons in the intact amphibian retina.

作者信息

Coleman P A, Miller R F

机构信息

Washington University School of Medicine, Department of Ophthalmology, St. Louis, Missouri 63110.

出版信息

J Neurophysiol. 1989 Jan;61(1):218-30. doi: 10.1152/jn.1989.61.1.218.

Abstract
  1. Whole-cell recordings have been obtained from intact, photoactive retinal neurons using patch-clamp electrodes in the amphibian superfused retina eyecup preparation. 2. After removal of the vitreous humor from the surface of the retina, using a collagenase with low tryptic activity, high-resistance seals (1-10 G omega) could be formed between the patch pipette and the cell membrane by applying mild suction to the pipette. Additional suction broke the membrane patch and provided continuity between the low-resistance pipette and the interior of the neuron. 3. Measurements of input resistance and time constant were obtained from bipolar, amacrine, and ganglion cells. Assuming the membrane capacitance was 1 microF/cm2, time constant data were used to derive the specific membrane resistance. The average specific membrane resistance for the inner retinal neurons in our sample was 68,000 omega.cm2. 4. Analysis of the charging curve induced by a brief current pulse applied to the soma was used to analyze the average electrotonic length of dendrites. The charging curves of some ganglion cells were well represented by a single exponential, suggesting that they were essentially isopotential. 5. The voltage decay along an equivalent cylinder model of a ganglion cell was calculated, using the experimentally obtained values of membrane resistance to compute decay of steady-state voltages along the dendritic tree. The calculations indicate that with the high membrane resistance values implied by this study, the electrotonic length of dendritic cables were short, and there may be relatively little attenuation of the synaptic potentials irrespective of their location along the dendritic tree.
摘要
  1. 在两栖动物视网膜眼杯灌流标本中,使用膜片钳电极从完整的、具有光活性的视网膜神经元上获得了全细胞记录。2. 在从视网膜表面去除玻璃体液后,使用具有低胰蛋白酶活性的胶原酶,通过对移液管施加轻微吸力,可在膜片移液管和细胞膜之间形成高电阻封接(1 - 10 GΩ)。额外的吸力会打破膜片,使低电阻移液管与神经元内部连通。3. 从双极细胞、无长突细胞和神经节细胞测量输入电阻和时间常数。假设膜电容为1 μF/cm²,时间常数数据用于推导比膜电阻。我们样本中视网膜内层神经元的平均比膜电阻为68,000 Ω·cm²。4. 对施加到胞体的短暂电流脉冲诱发的充电曲线进行分析,以分析树突的平均电紧张长度。一些神经节细胞的充电曲线可用单一指数很好地表示,表明它们基本上是等电位的。5. 使用实验获得的膜电阻值,计算神经节细胞等效圆柱体模型上的电压衰减,以计算沿树突树的稳态电压衰减。计算表明,根据本研究暗示的高膜电阻值,树突电缆的电紧张长度较短,并且无论突触电位沿树突树的位置如何,其衰减可能相对较小。

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