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大鼠中枢神经系统中甘氨酸转运体GLYT1和GLYT2的区域分布及发育变化

Regional distribution and developmental variation of the glycine transporters GLYT1 and GLYT2 in the rat CNS.

作者信息

Zafra F, Gomeza J, Olivares L, Aragón C, Giménez C

机构信息

Centro de Biología Molecular Severo Ochoa, Facultad de Ciencias, Universidad Autónoma de Madrid, Spain.

出版信息

Eur J Neurosci. 1995 Jun 1;7(6):1342-52. doi: 10.1111/j.1460-9568.1995.tb01125.x.

Abstract

The high-affinity glycine transporter in neurons and glial cells is the primary means of inactivating synaptic glycine. Previous molecular cloning studies have indicated heterogeneity of glycine transporters in the CNS. Here the distribution of glycine transporter GLYT1 and GLYT2 transcripts and proteins in different regions and developmental stages of the rat brain were analysed by Northern, Western and in situ hybridization techniques. Sequence-specific riboprobes and two specific antibodies raised against fusion proteins were used, containing either 76 or 193 amino acids of the C or N terminus of the GLYT1 and GLYT2 transporters respectively. High levels of GLYT1 transcripts were found in the spinal cord, brainstem and cerebellum, and moderate levels in forebrain regions such as the cortex or hippocampus. GLYT2 transcripts are restricted to the spinal cord, brainstem and cerebellum. The onset of both GLYT1 and GLYT2 expression in the brainstem occurred in late fetal life, and full expression of these proteins was observed before weaning. There was a stepwise increase in the levels of mRNA and protein for these two transporters, reaching a maximum by the second postnatal week, followed by a slight decrease until adult values were reached by the fourth postnatal week. These data reveal interesting parallelism between the distribution of different glycine transporters and glycine receptor subunits, and suggest discrete roles for distinct glycine transporters.

摘要

神经元和神经胶质细胞中的高亲和力甘氨酸转运体是使突触甘氨酸失活的主要方式。先前的分子克隆研究表明中枢神经系统中甘氨酸转运体存在异质性。在此,运用Northern、Western和原位杂交技术分析了大鼠脑不同区域和发育阶段中甘氨酸转运体GLYT1和GLYT2转录本及蛋白的分布情况。使用了序列特异性核糖探针以及两种针对融合蛋白产生的特异性抗体,这些融合蛋白分别包含GLYT1和GLYT2转运体C端或N端的76个或193个氨基酸。在脊髓、脑干和小脑中发现高水平的GLYT1转录本,而在诸如皮质或海马体等前脑区域中则为中等水平。GLYT2转录本局限于脊髓、脑干和小脑。脑干中GLYT1和GLYT2表达的起始发生在胎儿晚期,在断奶前观察到这些蛋白的完全表达。这两种转运体的mRNA和蛋白水平呈逐步上升,在出生后第二周达到峰值,随后略有下降,直至出生后第四周达到成年水平。这些数据揭示了不同甘氨酸转运体与甘氨酸受体亚基分布之间有趣的平行关系,并表明不同的甘氨酸转运体具有不同的作用。

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