Blanton M G, Kriegstein A R
Department of Neurology and Neurological Sciences, Stanford University School of Medicine, California 94305.
J Neurosci. 1991 Dec;11(12):3907-23. doi: 10.1523/JNEUROSCI.11-12-03907.1991.
We used loose-patch and whole-cell recording techniques to study the development of spontaneous action potential activity and spontaneous excitatory and inhibitory synaptic currents in embryonic neurons in the cerebral hemispheres of turtles. Sporadic action potential activity appeared early in development at stage 17, soon after morphologically identifiable pyramidal and nonpyramidal neurons were first observed in the cortex. As the cortical plate matured in midembryonic stages, action potential activity became more regular and fell into one of two distinct patterns, tonic and intermittent high-frequency firing. Spontaneous excitatory and inhibitory postsynaptic currents (EPSCs and IPSCs) appeared at developmental stages 18 and 20, respectively, after action potential activity was established. EPSCs and IPSCs exhibited characteristic ionic dependence and pharmacology throughout development. EPSCs reversed in direction at the equilibrium potential for cations and were sensitive to 6-cyano-7-nitroquinoxaline-2,3-dione, an antagonist of the non-NMDA type of glutamate receptor. IPSCs reversed at the equilibrium potential for chloride and were sensitive to bicuculline methiodide, a GABAA receptor antagonist. Spontaneous synaptic currents differed in their time course of development and in waveform parameters. Spontaneous synaptic currents differed in their time course of development and in waveform parameters. Spontaneous EPSCs appeared at stage 18 and increased progressively in frequency, from 0.2 +/- 0.1 Hz at stage 20 to 3.2 +/- 2.0 Hz at stage 26 (hatching), while spontaneous IPSCs appeared at stage 20 and surpassed EPSCs in frequency, increasing to 7.1 +/- 1.6 Hz at stage 26. EPSCs exhibited stable amplitudes during development, with a mean conductance of 126 +/- 20 pS at stage 26, while IPSCs increased in mean amplitude, from 180 +/- 12 pS at stage 18 to 260 +/- 44 pS at stage 26. The rise time to peak conductance of both types of synaptic currents increased with developmental time, for EPSCs increasing from 1.5 +/- 0.5 msec at stage 20 to 2.7 +/- 0.6 msec at stage 26 and for IPSCs increasing from 2.9 +/- 0.2 msec at stage 18 to 6.2 +/- 0.8 msec at stage 26. While the decay time constants increased for EPSCs, from 3.9 +/- 1.2 msec at stage 20 to 8.7 +/- 2.3 msec at stage 26, decay time constants for IPSCs showed a decreasing trend from 24.0 +/- 5.2 msec at stage 18 to 18.4 +/- 5.3 msec at stage 26. The excitatory and inhibitory synaptic currents were sensitive to the sodium channel blocker TTX and were thus dependent, in part, on spontaneous action potential activity.(ABSTRACT TRUNCATED AT 400 WORDS)
我们运用膜片钳和全细胞记录技术,研究了龟脑半球胚胎神经元中自发动作电位活动以及自发兴奋性和抑制性突触电流的发育情况。在胚胎发育早期的第17阶段,即首次在皮层中观察到形态可辨的锥体神经元和非锥体神经元后不久,就出现了零星的动作电位活动。随着胚胎中期皮层板的成熟,动作电位活动变得更加规律,并呈现出两种不同的模式之一,即紧张性发放和间歇性高频发放。自发兴奋性和抑制性突触后电流(EPSCs和IPSCs)分别在动作电位活动确立后的第18和20发育阶段出现。在整个发育过程中,EPSCs和IPSCs表现出独特的离子依赖性和药理学特性。EPSCs在阳离子平衡电位处发生方向反转,并且对非NMDA型谷氨酸受体拮抗剂6-氰基-7-硝基喹喔啉-2,3-二酮敏感。IPSCs在氯离子平衡电位处反转,并且对GABAA受体拮抗剂甲基荷包牡丹碱敏感。自发突触电流在发育的时间进程和波形参数方面存在差异。自发EPSCs在第18阶段出现,并在频率上逐渐增加,从第20阶段的0.2±0.1 Hz增加到第26阶段(孵化)的3.2±2.0 Hz,而自发IPSCs在第20阶段出现,且在频率上超过EPSCs,在第26阶段增加到7.1±1.6 Hz。EPSCs在发育过程中表现出稳定的幅度,在第26阶段平均电导为126±20 pS,而IPSCs的平均幅度增加,从第18阶段的180±12 pS增加到第26阶段的260±44 pS。两种类型突触电流达到峰值电导的上升时间都随着发育时间而增加,EPSCs从第20阶段的1.5±0.5毫秒增加到第26阶段的2.7±0.6毫秒,IPSCs从第18阶段的2.9±0.2毫秒增加到第26阶段的6.2±0.8毫秒。虽然EPSCs的衰减时间常数增加,从第20阶段的3.9±1.2毫秒增加到第26阶段的8.7±2.3毫秒,但IPSCs的衰减时间常数呈现出下降趋势,从第18阶段的24.0±5.2毫秒下降到第26阶段的18.4±5.3毫秒。兴奋性和抑制性突触电流对钠通道阻滞剂TTX敏感,因此部分依赖于自发动作电位活动。(摘要截选至400字)