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小鼠小脑中新型锌指基因wiz剪接形式的分子克隆及独特的发育表达模式

Molecular cloning and distinct developmental expression pattern of spliced forms of a novel zinc finger gene wiz in the mouse cerebellum.

作者信息

Matsumoto K, Ishii N, Yoshida S, Shiosaka S, Wanaka A, Tohyama M

机构信息

Division of Structural Cell Biology, Nara Institute of Science and Technology (NAIST), 8916-5 Takayama, Ikoma, Nara 630-0101, Japan.

出版信息

Brain Res Mol Brain Res. 1998 Oct 30;61(1-2):179-89. doi: 10.1016/s0169-328x(98)00216-2.

Abstract

In the course of a study conducted to identify the mouse homologue of Drosophila eyes absent (eya), we isolated a novel mouse cDNA fragment which show little homology to eya but encodes a protein with Krüppel (C2H2)-type zinc finger motifs. By further screening using this cDNA fragment as a probe, we obtained the short and long forms of full-length cDNAs, which were apparently alternatively spliced products from one gene. Since both mRNAs encode proteins with widely-interspaced zinc finger motifs, we termed this gene wiz and refer to the short and long wiz transcripts as wizS and wizL, respectively. In situ hybridization studies using the probe against the region common to wizS and wizL showed that these mRNAs were expressed abundantly in the granule cell layers of the mouse cerebellum, the olfactory bulb, and the dentate gyrus, whereas the same technique using the probe against only wizL could not detect positive signals in the developing cerebellum, indicating that there is no expression of wizL mRNA there. Northern blot and in situ hybridization analyses demonstrated that the extracerebellar regions expressed both wizS and wizL mRNAs from the midgestational period to adulthood. The finding that two types of wiz transcripts (wizS and wizL) are expressed with different developmental patterns might indicate separate transcription functions in the cerebellar granule cells and the extracerebellar regions.

摘要

在一项旨在鉴定果蝇无眼(eya)的小鼠同源物的研究过程中,我们分离出一个新的小鼠cDNA片段,该片段与eya的同源性很低,但编码一种具有克虏伯(C2H2)型锌指基序的蛋白质。通过以该cDNA片段为探针进行进一步筛选,我们获得了全长cDNA的短形式和长形式,它们显然是来自一个基因的可变剪接产物。由于这两种mRNA都编码具有广泛间隔锌指基序的蛋白质,我们将这个基因命名为wiz,并将短和长的wiz转录本分别称为wizS和wizL。使用针对wizS和wizL共有的区域的探针进行原位杂交研究表明,这些mRNA在小鼠小脑的颗粒细胞层、嗅球和齿状回中大量表达,而使用仅针对wizL的探针的相同技术在发育中的小脑中未检测到阳性信号,表明那里没有wizL mRNA的表达。Northern印迹和原位杂交分析表明,从妊娠中期到成年期,小脑外区域表达wizS和wizL两种mRNA。两种类型的wiz转录本(wizS和wizL)以不同的发育模式表达这一发现可能表明在小脑颗粒细胞和小脑外区域存在不同的转录功能。

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