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首页> 外文期刊>Neurochemistry International: The International Journal for the Rapid Publication of Critical Reviews, Preliminary and Original Research Communications in Neurochemistry >Functional and molecular analysis of glutamate-gated channels by patch-clamp and RT-PCR at the single cell level.
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Functional and molecular analysis of glutamate-gated channels by patch-clamp and RT-PCR at the single cell level.

机译：通过膜片钳和RT-PCR在单个细胞水平上对谷氨酸门控通道进行功能和分子分析。

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摘要

In the central nervous system (CNS) rapid excitatory neurotransmission is mainly mediated by ligand gated, cationic channels activated by glutamate. Three main subtypes of glutamate-gated channels have been characterized by pharmacological studies. They have been named according to their preferred agonist, N-methyl-D-aspartate (NMDA), high affinity kainate and alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA). Furthermore, a large diversity within each class of glutamate-gated channels has been revealed by the molecular cloning of multiple subunits and their spliced and edited variants (for review see Wisden and Seeburg, 1993). These subunits can potentially form different oligomeric complexes with diverging properties. A crucial question is therefore to determine the actual subunit composition of naturally occurring glutamate receptors. We have combined patch-clamp recording, reverse transcription (RT) and PCR to correlate, at the single cell level, the pattern of subunits expression with the functional properties of native glutamate receptors. We describe here results obtained on the AMPA receptors of hippocampal neurones and on the NMDA receptors of cerebellar granule cells which show that the subunit composition of these two types of receptors explains some of their functional properties. Furthermore, our data also indicate that the expression of NMDA receptor subunits during the postnatal development of cerebellar granule cells is regulated by an activity-dependent mechanism.
机译：在中枢神经系统（CNS）中，快速兴奋性神经传递主要由被谷氨酸激活的配体门控的阳离子通道介导。谷氨酸门控通道的三种主要亚型已通过药理学研究进行了表征。它们根据其优选的激动剂命名，即N-甲基-D-天冬氨酸（NMDA），高亲和力的海藻酸酯和α-氨基-3-羟基-5-甲基-4-异恶唑丙酸酯（AMPA）。此外，通过对多个亚基及其剪接和编辑的变体进行分子克隆，揭示了在谷氨酸门控通道的每一类中的巨大多样性（综述参见Wisden和Seeburg，1993）。这些亚基可能形成具有不同性质的不同寡聚复合物。因此，关键的问题是确定天然存在的谷氨酸受体的实际亚基组成。我们结合了膜片钳记录，逆转录（RT）和PCR，以在单个细胞水平上将亚基表达模式与天然谷氨酸受体的功能特性相关联。我们在这里描述了在海马神经元的AMPA受体和小脑颗粒细胞的NMDA受体上获得的结果，这些结果表明这两种受体的亚基组成解释了它们的某些功能特性。此外，我们的数据还表明，小脑颗粒细胞产后发育过程中NMDA受体亚基的表达受活性依赖机制的调节。

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