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在单个海马突触处对单个量子进行的膜片钳记录。

Loose-patch recordings of single quanta at individual hippocampal synapses.

作者信息

Forti L, Bossi M, Bergamaschi A, Villa A, Malgaroli A

机构信息

Department of Biological and Technological Research, DIBIT, Scientific Institute San Raffaele, Milano, Italy.

出版信息

Nature. 1997 Aug 28;388(6645):874-8. doi: 10.1038/42251.

Abstract

Synapses in the central nervous system are typically studied by recording electrical responses from the cell body of the postsynaptic cell. Because neurons are normally connected by multiple synaptic contacts, these postsynaptic responses reflect the combined activity of many thousands synapses, and it remains unclear to what extent the properties of individual synapses can be deduced from the population response. We have therefore developed a method for recording the activity of individual hippocampal synapses. By capturing an isolated presynaptic bouton inside a loose-patch pipette and recording from the associated patch of postsynaptic membrane, we were able to detect miniature excitatory postsynaptic currents ('minis') arising from spontaneous vesicle exocytosis at a single synaptic site, and to compare these with minis recorded simultaneously from the cell body. The average peak conductance at a single synapse was about 900 pS, corresponding roughly to the opening of 90 AMPA-type glutamate-receptor channels. The variability in this conductance was about 30%, matching the value reported for the neuromuscular junction. Given that our synapses displayed single postsynaptic densities (PSDs), this variability is larger than would be predicted from the random opening of receptor channels, suggesting that they are not saturated by the content of a single vesicle. Therefore the response to a quantum of neurotransmitter at these synapses is not limited by the number of available postsynaptic receptors.

摘要

中枢神经系统中的突触通常是通过记录突触后细胞胞体的电反应来进行研究的。由于神经元通常通过多个突触连接,这些突触后反应反映了成千上万个突触的综合活动,而从群体反应中能在多大程度上推断出单个突触的特性仍不清楚。因此,我们开发了一种记录单个海马突触活动的方法。通过将一个孤立的突触前终扣捕获在一个 loose-patch 移液管内,并从相关的突触后膜片进行记录,我们能够检测到来自单个突触位点自发囊泡胞吐产生的微小兴奋性突触后电流(“微小电流”),并将其与同时从胞体记录的微小电流进行比较。单个突触的平均峰值电导约为900 pS,大致相当于90个AMPA型谷氨酸受体通道的开放。这种电导的变异性约为30%,与报道的神经肌肉接头的值相匹配。鉴于我们的突触显示出单个突触后致密物(PSD),这种变异性比从受体通道随机开放所预测的要大,这表明它们不会被单个囊泡的内容物饱和。因此,这些突触对一个神经递质量子的反应不受可用突触后受体数量的限制。

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