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Molecular anatomical research of nuclear membrane in mitotic phase : the ultra structure of the surface and genes regulate it.

有丝分裂期核膜的分子解剖学研究：表面超微结构和基因调控。

基本信息

批准号：
18590188
负责人：
HIROSE Eiji
金额：
$ 2.43万
依托单位：
Kyushu University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18590188/
关键词：
Anatomy Cell Histology Ultrastructure of the cell Genetics G protein Xenonus laevis Xenopus leavis

项目摘要

For the first step of investigating the functions of Rag gene family molecules in nuclear division process, we isolated the homologues of Rag gene from African clawed frog, Xenopus laevis (XRag1 and XRag2). A multiple alignment and phylogenetic tree indicated that all eukaryotes carry two closely related genes. Together with binding of these two gene products in human and yeast previously reported, XRag1 and XRag2 could form the minimal functional complex in vivo. However, a level of XRag1 and XRag2 transcripts were not linked revealed by RT-PCR. XRag2 expression showed rapid decrease after Mid Blastula Transition (MBT) in contrast to the almost constant level of XRag2. Then, we confirmed the spatial expression patterns of two genes by in situ hybridization. In early stages before MBT, both XRag1 and XRag2 are expressed in animal hemisphere but XRag2 is decreased after stage 8 and no longer detectable in stage 12. Simultaneous expression of XRag1 and XRag2 was detected again after stage 25, tail bud stage in neuronal tissues (eye and ear primordium). In summary, (1) XRag1 and XRag2 shared incompatible genetic functions. (2) During stage 10-25, only XRag1 is expressed. This suggests XRag1 homodimer (or homooligomer) is the functional unit in these stages. (3) Thus, expression ratio of XRag1 and XRag2 may regulate the switching of the cell division mode at MBT. (4) In later stages, these genes are specifically expressed in neuronal tissues, suggesting new XRag1 and XRag2 roles in the development of peripheral nervous system.

为了研究Rag基因家族分子在细胞核分裂过程中的功能，我们从非洲爪蟾（Xenopus laevis）中分离出Rag基因的同源分子（XRag1和XRag2）。多重比对和系统发育树表明，所有真核生物都携带两个密切相关的基因。XRag1和XRag2与这两个基因产物结合在人体内和酵母中，可以在体内形成最小的功能复合物。然而，通过RT-PCR发现XRag1和XRag2转录本的水平没有关联。在囊胚中期转化（MBT）后，XRag2的表达迅速下降，而XRag2的表达基本保持不变。然后，我们通过原位杂交确定了两个基因的空间表达模式。在MBT前的早期，XRag1和XRag2在动物半球均有表达，但XRag2在第8期后减少，在第12期不再检测到。XRag1和XRag2在25期（尾芽期）神经元组织（眼和耳原基）中同时表达。综上所述，(1)XRag1和XRag2具有不相容的遗传功能。(2)第10 ~ 25期仅表达XRag1。这表明XRag1同聚二聚体（或同聚物）是这些阶段的功能单位。(3)因此，XRag1和XRag2的表达比例可能调控MBT细胞分裂模式的切换。(4)在后期，这些基因在神经组织中特异性表达，提示XRag1和XRag2在周围神经系统的发育中有新的作用。