Development of A New Method to Investigate Cell Migration in the Developing Central Nervous System

开发一种研究发育中中枢神经系统细胞迁移的新方法

• 批准号: 09680740
• 负责人: TAMAMAKI Nobuaki
• 金额: $ 2.05万
• 依托单位: Department of Anatomy, Fukui Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 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-09680740/
• 关键词: Migration; Migration speed; Olfactory Buld; Thymin-dimer; UV-light; Fiber Optic Cable; チミンダイマー; 紫外線照射; 色素性乾皮症; ミュータントマウス; 光ファイバー

We have devised a novel method to study cell migration in vitro and in vivo. The new method involves UV-irradiating cells through a fiber optic cable and subsequently identifying irradiated cells by virtue of the formation of thymine-dimer in the nuclei. We applied the new method to investigate migration of neuronal precursor cells in the rostral migratory stream in the neonatal rat olfactory bulb. In vitro, UV-irradiation for one second through the fiber optic cable allowed formation of enough thymine-dimers to be detected immunohistochemically after 6 hours of incubation. A significant proportion of irradiated cells continued to migrate in the same direction at the same speed as before irradiation. There was no significant difference in the cell-migration distance at 6 hours with or without the UV-irradiation in vitro. We applied this UV-thymine-dimer labeling method to study cell migration in the olfactory bulb in vivo, With this method, we could find that a small number of cells migrated also caudally. Three times more cells in the SV of the olfactory bulb migrated rostrally, but there was also a significant extent of caudally directed cell migration. The new method also allowed us to measure the speed of cell-migration, which was estimated to be about 70 ? m/hr to the rostral direction and 37 ? m/hr to the caudal direction, at maximum.

我们设计了一种新的方法来研究细胞在体内和体外的迁移。这种新方法是通过光纤对细胞进行紫外线照射，然后通过细胞核中胸腺嘧啶二聚体的形成来识别被照射的细胞。我们应用新方法研究了新生大鼠嗅球吻侧迁移流中神经元前体细胞的迁移。在体外，通过光缆进行一秒钟的紫外线照射，可以形成足够的胸腺嘧啶二聚体，在6小时的孵育后进行免疫组织化学检测。很大一部分受辐照的细胞继续以与辐照前相同的方向和速度迁移。体外辐照前后6小时细胞迁移距离差异无统计学意义。我们将这种紫外胸腺嘧啶二聚体标记方法应用于活体嗅球中细胞迁移的研究，通过这种方法，我们可以发现少量细胞也在尾端迁移。嗅球SV中有三倍多的细胞向喙侧迁移，但也有显著程度的尾侧定向细胞迁移。新方法还允许我们测量细胞迁移的速度，估计大约为70 ？每小时每小时到东南方向和37 ？M /hr到尾端方向，最大。

项目成果

Tamamaki,N.et al.: "Origin and route of tangentially migrating neurons in the developing neocorteical intermediate zone." Journal of Neuroscience. 17. 8313-8323 (1997)

Tamamaki,N.et al.：“发育中的新皮质中间区切向迁移神经元的起源和路线。”

Nobuaki Tamamaki, Yoshihiko Sugimoto, Kiyoji Tanaka, and Rumiko Takauji: "Tangential cell migration in the neocortex monitored by marking nuclei with UV-irradiation via a fiber optic cable." Development. (submitted). (1999)

Nobuaki Tamamaki、Yoshihiko Sugimoto、Kiyoji Tanaka 和 Rumiko Takauji：“通过光纤电缆用紫外线照射标记细胞核来监测新皮质中的切向细胞迁移。”

Tamamaki N.et al.: "The development of afferent fiber lamination in the infrapyramidal Blade of the rat dentate gyrus." J.Comp.Neurol.in press.

Tamamaki N.等人：“大鼠齿状回锥体下叶片中传入纤维层压的发展。”

Tamamaki N.et al.: "Tangential cell migration in the neocortex monitored by marking nuclei with UV-irradiation via a fiber optic cable." Development. submitted.

Tamamaki N.等人：“通过光纤电缆用紫外线照射标记细胞核来监测新皮质中的切向细胞迁移。”

Tamamaki,N.et al.: "Intermediate zone and intermediate zone neurons : Do IZ neurons pioneer the layer V neurons in rats?." Developmental Neuroscience. 19. 112-116. ((1997))

Tamamaki,N.等人：“中间区和中间区神经元：IZ 神经元是否是大鼠中 V 层神经元的先驱？”