Functional analyses of neural development genes

神经发育基因的功能分析

• 批准号: 12210003
• 负责人: MASU Masayuki
• 金额: $ 69.7万
• 依托单位: University of Tsukuba
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research on Priority Areas
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12210003/
• 关键词: Neural development; Sulfatase; Heparan sulfate; Autotaxin; Floor plate; Knockout mouse; オートタキシン; 神経回路; シグナル伝達; 神経発生; 遺伝子破壊; トランスジェニックマウス; サルファターゼ; 標的遺伝子破壊

This project aims to clarify the molecular mechanism that regulates neural development. For this purpose, we characterized the SulfFP1, SulfFP2, and Autotaxin genes that we have previously identified as floor plate specific genes. We summarize our findings as follows. SulfFP is a novel member of the sulfatase family, it is secreted into culture medium when transfected, it is cleaved in furin-dependent and independent manners, it has endosulfatase activity that hydrolyses 6-0-sulfate in the glucosamine residues in intact heparin and heparan sulfate, trisulfated disaccharide units becomes a good substrate for SulfFPs, and its activity is high at neutral pH rather than acidic pH. Autotaxin is a secreted protein that was identified as a chemotaxis-promoting factor. The presence of a phosphodiesterase domain predicted the role of Autotaxin in the nucleotide metabolism in the extracellular space. Recent studies, however, revealed that Autotaxin has a lysophospholipase D activity that generat … More es lysophosphatidic acid (LPA) and sphingosine-1-phosphate (SIP).Although many lines of in vitro studies have provided ample evidence suggesting their functions, their physiological roles in vivo remain unknown. Therefore, we have generated the knockout mice for these genes and analyzed their phenotypes. The mice deficient for either SulfFP1 or SulfFP2 genes appear to be normal, suggesting the functional redundancy of these genes. Indeed our previous studies showed that SulfFP1 and SulfFP2 expressions are largely overlapping in mouse embryos. In contrast, many of the mice deficient for both SulfFP1 and SulfFP2 genes die within 1 day after birth for unknown reason. The double-knockout mice showed abnormality in the nervous system and growth retardation. Autotaxin-deficient mice die at embryonic day 8~9, and some mice showed excencephaly phenotype, indicating the importance of Autotaxin in neural development. Conditional knockout mice that have defect only in the nervous system are necessary to investigate the role of Autotaxin in late neural development. Less

该项目旨在阐明调节神经发育的分子机制。为此，我们鉴定了SulfFP1、SulfFP2和Autoaxin基因，这些基因我们以前已经确定为底板特异性基因。我们将我们的发现总结如下。SulfFP是硫酸盐酶家族中的一个新成员，它在转染后分泌到培养基中，以依赖于呋喃的和独立的方式被切割，它具有内切酶活性，可以降解完整肝素和肝素硫酸盐中氨基葡萄糖残基中的6-0-硫酸盐，三硫化二糖单元是硫酸盐FP的良好底物，并且在中性pH而不是酸性pH下活性较高。自体趋化蛋白是一种分泌型蛋白，被认为是一种趋化促进因子。磷酸二酯酶结构域的存在预测了自体趋化蛋白在细胞外空间核苷酸代谢中的作用。然而，最近的研究表明，自体趋化蛋白具有产生…的溶血磷脂酶D活性更多的ES溶血磷脂酸(LPA)和鞘氨醇-1-磷酸(SIP)。尽管许多体外研究已经提供了充分的证据表明它们的功能，但它们在体内的生理作用仍不清楚。因此，我们产生了这些基因的敲除小鼠，并分析了它们的表型。缺乏SulfFP1或SulfFP2基因的小鼠似乎是正常的，这表明这些基因的功能冗余。事实上，我们之前的研究表明，SulfFP1和SulfFP2在小鼠胚胎中的表达在很大程度上是重叠的。相比之下，许多同时缺乏SulfFP1和SulfFP2基因的小鼠在出生后1天内死亡，原因不明。双基因敲除小鼠表现出神经系统异常和生长迟缓。自体趋化蛋白缺陷小鼠在胚胎第8~9天死亡，部分小鼠表现为异常脑型，提示自体趋化蛋白在神经发育中的重要性。只有神经系统有缺陷的条件性基因敲除小鼠是必要的，以研究自体趋化蛋白在神经发育后期的作用。较少

项目成果

Potentiation of capsaicin receptor activity by metabotropic ATP receptors as a possible mechanism for ATP-evoked pain and hyperalgesia

• DOI: 10.1073/pnas.111025298
• 发表时间: 2001-05
• 期刊: Proceedings of the National Academy of Sciences of the United States of America
• 影响因子: 11.1
• 作者: M. Tominaga;M. Wada;M. Masu

What do we know about the brain? (ed. Ihara Y)

我们对大脑了解多少？

• 发表时间: 2005
• 作者: Kato M;Seki N;Sugano S;Hashimoto K;Masuho Y;Muramatsu M;Kaibuchi K;Nakafuku M.;Masu M.et al.
• 通讯作者: Masu M.et al.

Sato et al.: "Distribution of cystine/glutamate exchange transporter, system Xc ; in the mouse brain"Journal of Neuroscience. 22. 8028-8033 (2002)

Sato 等人：“胱氨酸/谷氨酸交换转运蛋白系统 Xc 的分布；在小鼠大脑中”神经科学杂志。

Tsukada, S.et al.: "Activation of protein kinase A by nitric oxide in cultured dorsal root ganglion neurites of the rat, examined ……"Neuroscience Letters. 318. 17-20 (2002)

Tsukada, S. 等人：“在培养的大鼠背根神经节神经突中一氧化氮激活蛋白激酶 A，已检测......”神经科学快报。 318. 17-20 (2002)

桝正幸: "神経分化と神経回路構築の分子機構"実験医学. 18・19. 2636-2643 (2000)

Masayuki Masu：“神经分化和神经回路构建的分子机制”实验医学18・19（2000）。