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鱼类中枢神经系统发育和再生过程中α1-微管蛋白基因表达的诱导

Induction of alpha1-tubulin gene expression during development and regeneration of the fish central nervous system.

作者信息

Hieber V, Dai X, Foreman M, Goldman D

机构信息

Mental Health Research Institute and Department of Biological Chemistry, University of Michigan, Ann Arbor 48109, USA.

出版信息

J Neurobiol. 1998 Nov 15;37(3):429-40. doi: 10.1002/(sici)1097-4695(19981115)37:3<429::aid-neu8>3.0.co;2-n.

Abstract

The alpha1- and alpha2-tubulin encoding genes were cloned from a goldfish genomic DNA library. alpha1- and alpha2-tubulin RNA expression was examined in developing and adult retinas. These studies demonstrated increased alpha1-tubulin RNA in presumptive ganglion cells that grow axons early in retinal development and in adult retinal ganglion cells whose optic axons had been damaged. The alpha2-tubulin RNA was undetectable in developing retina and constitutively expressed in adult retinal ganglion cells regardless of optic nerve crush. To determine if these changes in alpha1-tubulin RNA reflected changes in alpha1-tubulin promoter activity, we introduced into zebrafish embryos and adult goldfish retinal explants expression vectors harboring the alpha1-tubulin gene's promoter. These studies showed that the alpha1-tubulin promoter confers a developmentally regulated, neuron-restricted pattern of reporter gene expression in vivo and its activity is increased in adult retinal neurons induced to regenerate their axons. Promoter deletions defined regions of alpha1-tubulin DNA necessary for this pattern of expression. These results suggest that DNA sequences necessary for alpha1-tubulin gene induction during central nervous system development and regeneration are contained within the alpha1-tubulin gene's 5'-flanking DNA and that this promoter will be useful for identifying these elements and their DNA binding proteins.

摘要

从金鱼基因组DNA文库中克隆了α1 - 和α2 - 微管蛋白编码基因。检测了发育中和成年视网膜中α1 - 和α2 - 微管蛋白RNA的表达。这些研究表明,在视网膜发育早期生长轴突的假定神经节细胞以及视神经轴突受损的成年视网膜神经节细胞中,α1 - 微管蛋白RNA增加。在发育中的视网膜中未检测到α2 - 微管蛋白RNA,并且在成年视网膜神经节细胞中持续表达,无论视神经是否受到挤压。为了确定α1 - 微管蛋白RNA的这些变化是否反映了α1 - 微管蛋白启动子活性的变化,我们将携带α1 - 微管蛋白基因启动子的表达载体导入斑马鱼胚胎和成年金鱼视网膜外植体中。这些研究表明,α1 - 微管蛋白启动子在体内赋予报告基因表达一种发育调控的、神经元限制的模式,并且在诱导轴突再生的成年视网膜神经元中其活性增加。启动子缺失确定了这种表达模式所需的α1 - 微管蛋白DNA区域。这些结果表明,中枢神经系统发育和再生过程中α1 - 微管蛋白基因诱导所需的DNA序列包含在α1 - 微管蛋白基因的5' - 侧翼DNA中,并且该启动子将有助于鉴定这些元件及其DNA结合蛋白。

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