Network formation of motoneurons

运动神经元的网络形成

• 批准号: 09480207
• 负责人: TANAKA Hideaki
• 金额: $ 8.51万
• 依托单位: Kumamoto University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09480207/
• 关键词: Motoneuron; network formation; Eph; receptor tyrosine kinase; ephrin; axonal guidance; Eph受容体型チロシンキナーゼ; 阻害モノクローナル抗体; 軸索成長; ガイダンス

We previously found that two of the Eph receptor tyrosine kinases, EphA3 and EphA4, are transiently expressed in subsets of chick embryonic motoneurons and that the Eph ligands, ephrin-A2 and ephrin-A5, inhibit neurite outgrowth from the motoneurons in culture. Here we developed functional blocking monoclonal antibodies against EphA3, EphA4 and ephrin-A2 in order to examine their functional roles in axonal guidance of motoneurons in vivo. We raised many monoclonal antibodies against the chimeric proteins of each extracellular domain and Fc region of human IgG.Among them some antibodies had blocking activity for ligand-receptor binding and functional blocking activities in stripe assay of retinotectal system. The epitopes of anti-EphA4 and anti-ephrin-A2 were determined to be localized at cysteine rich domain and N-terminal region, respectively. These domains appeared to be essential for Eph receptor-ligand binding. We are now examining the effects of these functional blocking antibodies on motoneuron axonal growth in chick embryos.

我们以前发现，两个Eph受体酪氨酸激酶，EphA 3和EphA 4，是瞬时表达的鸡胚运动神经元的亚群和Eph配体，ephrin-A2和ephrin-A5，抑制神经突起生长的运动神经元在文化。在这里，我们开发了功能性阻断单克隆抗体EphA 3，EphA 4和ephrin-A2，以检查其功能的作用，在轴突的运动神经元在体内的指导。我们制备了多株抗人IgG各胞外区和Fc区嵌合蛋白的单克隆抗体，其中部分抗体具有配体-受体结合阻断活性和视网膜顶盖系统条纹试验功能阻断活性。抗EphA 4和抗Ephrin-A2的表位分别位于富含半胱氨酸的结构域和N-末端区域。这些结构域似乎是必不可少的Eph受体-配体结合。我们现在正在研究这些功能性阻断抗体对鸡胚运动神经元轴突生长的影响。

项目成果

Koo S.J.: "species-specific immunostaing of embryonic corneal nerves." Journal of Neuroscience Methods. 85・1. 63-71 (1998)

Koo S.J.：“胚胎角膜神经的物种特异性免疫染色”。《神经科学方法杂志》85・1（1998）。

藤井智子: "細胞接着蛋白質、SCI" 蛋白質 核酸 酵素. 42・3. 552-554 (1997)

藤井智子：“细胞粘附蛋白，SCI”蛋白质核酸酶42・3（1997）。

Monschau, B.: "Shared and distinct functions of RAGS and ELF-1 in guiding retinal axons." EMBO J.16 : 6. 1258-1267 (1997)

Monschau, B.：“RAGS 和 ELF-1 在引导视网膜轴突方面具有共同和不同的功能。”

Ohta, K.: "The inhibitory effect on neurite outgrowth of motoneurons exerted by the ligands ELF-1 and RAGS." Mech.Dev.64 : 1-2. 127-135 (1997)

Ohta, K.：“配体 ELF-1 和 RAGS 对运动神经元神经突生长的抑制作用。”

Hirano Shigeki: "Normal ontogenic observations on the expression of Eph receptor tyrosine kinase,Cek8,in chick embryos." Anatomy & Embryology. 197・3. 187-197 (1998)

Hirano Shigeki：“鸡胚胎中 Eph 受体酪氨酸激酶 Cek8 表达的正常个体发育观察”197・3（1998）。