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首页> 外文期刊>The European Journal of Neuroscience >N-ethylmaleimide increases release probability at GABAergic synapses in layer I of the mouse visual cortex.
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N-ethylmaleimide increases release probability at GABAergic synapses in layer I of the mouse visual cortex.

机译：N-乙基马来酰亚胺增加了小鼠视觉皮层I层中GABA能突触的释放可能性。

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摘要

The sulphydryl alkylating agent N-ethylmaleimide (NEM) has been often used as an uncoupler of pertussis toxin-sensitive G-proteins. However, the effects of NEM on gamma-aminobutyric acid (GABA)ergic synaptic transmission remain controversial. Using the whole-cell patch-clamp technique, GABA(A) receptor-mediated postsynaptic currents (IPSCs) have been recorded from Cajal-Retzius (CR) cells in layer I of the neonatal mouse visual cortex. NEM increased the frequencies of both spontaneous and miniature IPSCs (mIPSCs) without an effect on the median mIPSC amplitudes or mIPSC kinetics. The NEM actions on mIPSCs did not depend on the extracellular Ca(2+), Ca(2+) release from intracellular stores, adenylyl cyclase and protein kinase A activities. NEM increased the mean amplitudes of evoked IPSCs and strongly decreased the paired-pulse ratio. The size of the readily releasable pool of presynaptic vesicles (RRP) was estimated using a high-frequency stimulation protocol. The RRP size was not affected by NEM. In addition, NEM significantly decreased the latency between the stimulus and the onset of GABA release. These results suggest that NEM selectively increases GABA release probability. At postnatal day 2, mIPSCs were observed only in about 30% of CR cells. NEM application revealed, however, that more than 90% of CR cells receive GABAergic inputs. Therefore, NEM seems to be a useful tool to verify the existence of 'silent' GABAergic synapses.
机译：巯基烷基化剂N-乙基马来酰亚胺（NEM）通常被用作百日咳毒素敏感G蛋白的解偶联剂。然而，NEM对γ-氨基丁酸（GABA）能量突触传递的影响仍存在争议。使用全细胞膜片钳技术，已从新生小鼠视觉皮层I层的Cajal-Retzius（CR）细胞中记录了GABA（A）受体介导的突触后电流（IPSC）。 NEM增加了自发和微型IPSC（mIPSC）的频率，而没有影响中值mIPSC幅度或mIPSC动力学。 NEM对mIPSC的作用并不取决于细胞外Ca（2 +），Ca（2+）从细胞内存储，腺苷酸环化酶和蛋白激酶A的活性释放。 NEM增加了诱发IPSC的平均幅度，并大大降低了配对脉冲比。突触前囊泡（RRP）的容易释放池的大小使用高频刺激协议进行了估计。 RRP大小不受NEM的影响。此外，NEM显着减少了刺激与GABA释放开始之间的潜伏期。这些结果表明，NEM有选择地增加了GABA释放的可能性。出生后第2天，仅在约30％的CR细胞中观察到mIPSC。 NEM应用显示，但是，超过90％的CR细胞接受GABA能输入。因此，NEM似乎是验证“沉默的” GABA能突触的存在的有用工具。

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