Tyzio Roman, Ivanov Anton, Bernard Cristophe, Holmes Gregory L, Ben-Ari Yehezkiel, Khazipov Roustem
Institut de la Neurobiologie de la Méditterranée-Institute National de la Santé et de la Recherche Médicale U29, Marseille, France.
J Neurophysiol. 2003 Nov;90(5):2964-72. doi: 10.1152/jn.00172.2003. Epub 2003 Jul 16.
A depolarized resting membrane potential has long been considered to be a universal feature of immature neurons. Despite the physiological importance, the underlying mechanisms of this developmental phenomenon are poorly understood. Using perforated-patch, whole cell, and cell-attached recordings, we measured the membrane potential in CA3 pyramidal cells in hippocampal slices from postnatal rats. With gramicidin perforated-patch recordings, membrane potential was -44 +/- 4 (SE) mV at postnatal days P0-P2, and it progressively shifted to -67 +/- 2 mV at P13-15. A similar developmental change of the membrane potential has been also observed with conventional whole cell recordings. However, the value of the membrane potential deduced from the reversal potential of N-methyl-d-aspartate channels in cell-attached recordings did not change with age and was -77 +/- 2 mV at P2 and -77 +/- 2 mV at P13-14. The membrane potential measured using whole cell recordings correlated with seal and input resistance, being most depolarized in neurons with high, several gigaohms, input resistance and low seal resistance. Simulations revealed that depolarized values of the membrane potential in whole cell and perforated-patch recordings could be explained by a shunt through the seal contact between the pipette and membrane. Thus the membrane potential of CA3 pyramidal cells appears to be strongly negative at birth and does not change during postnatal development.
去极化的静息膜电位长期以来一直被认为是未成熟神经元的一个普遍特征。尽管其具有生理重要性,但这种发育现象的潜在机制却知之甚少。我们使用穿孔膜片钳、全细胞和细胞贴附记录技术,测量了新生大鼠海马切片中CA3锥体神经元的膜电位。使用短杆菌肽穿孔膜片钳记录时,出生后第0 - 2天的膜电位为-44±4(标准误)mV,在出生后第13 - 15天逐渐转变为-67±2 mV。在传统全细胞记录中也观察到了类似的膜电位发育变化。然而,从细胞贴附记录中N - 甲基 - D - 天冬氨酸通道的反转电位推导出的膜电位值并不随年龄变化,在出生后第2天为-77±2 mV,在出生后第13 - 14天为-77±2 mV。使用全细胞记录测量的膜电位与封接电阻和输入电阻相关,在具有高输入电阻(数吉欧)和低封接电阻的神经元中膜电位最去极化。模拟结果表明,全细胞和穿孔膜片钳记录中膜电位的去极化值可以通过移液器与细胞膜之间封接接触处的分流来解释。因此,CA3锥体神经元的膜电位在出生时似乎就非常负,并且在出生后发育过程中没有变化。