Application of the whole embryo culture system to the study on the brain development

全胚胎培养系统在脑发育研究中的应用

• 批准号: 08458261
• 负责人: KAWANO Hitoshi
• 金额: $ 0.83万
• 依托单位: KEIO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08458261/
• 关键词: mouse; midbrain; whole embryo culture; development; L1; phosphacan; NCAM-H; アンチセンスDNA; 中脳黒質; ドーパミンニューロン; 放射状グリア; テネイシン; チロシン水酸化酵素

In spite of the great advance of the gene technology in recent years, the mechanism of mammalian brain development is not fully known. One of the reasons is the difficulty in observing and manipulating mammalian embryos which develop in the mother's uterus. In the present study, some experimental manupilations were made to the embryonic mouse brain by using the whole embryo culture system to clarify the mechanisms of the brain development. We have already studied the in vivo development of the mouse midbrain and obtained the data which indicate that molecular interactions between neural cell adhesion molecules, L1 and a brain-specific chondroitin sulfate proteoglycan, phosphacan, are involved in the migration of mesencephalic dopamine-containing neurons (Kawano et al., 1997 ; Ohyama et al., 1998 in press). Since L1 have been reported to bind phosphacan in vivo, molecular interaction between the two molecules may play important roles in the migration of dopamine neurons. Based on these observation, we made microinjection of an enzyme which degrades chondrotin sulfate and antibody ageinst L1 into the ventricle of E13 mouse embryos using the whole embryo culture system, and exmined the brain histologically. This system will be a useful tool to study the developmental mechanism of the mammalian brain.

尽管近年来基因技术取得了巨大进步，但哺乳动物大脑发育的机制尚不完全清楚。原因之一是难以观察和操作在母亲子宫中发育的哺乳动物胚胎。本研究利用全胚胎培养系统对小鼠胚胎大脑进行了实验操作，以阐明大脑发育的机制。我们已经研究了小鼠中脑的体内发育，并获得了数据表明神经细胞粘附分子L1和大脑特异性硫酸软骨素蛋白聚糖磷酸聚糖之间的分子相互作用参与了中脑含多巴胺神经元的迁移（Kawano 等人，1997 年；Ohyama 等人，1998 年出版）。由于 L1 据报道在体内结合磷酸聚糖，因此两个分子之间的分子相互作用可能在多巴胺神经元的迁移中发挥重要作用。基于这些观察，我们使用整个胚胎培养系统将降解硫酸软骨素的酶和抗体ageinst L1显微注射到E13小鼠胚胎的心室中，并对脑进行组织学检查。该系统将成为研究哺乳动物大脑发育机制的有用工具。

项目成果

Ikawa H,Kawano H,Takeda Y,Masuyama H,Watanabe K,Endo M,Yokoyama J,Kitajima M,Uyemura K,Kawamura K: "Impaired expression of neural cell adhesion molecule L1 in the extrinsic nerve fibers in Hirschsprung's disease." J.Ped.Surg.32. 542-545 (1997)

Ikawa H、Kawano H、Takeda Y、Masuyama H、Watanabe K、Endo M、Yokoyama J、Kitajima M、Uyemura K、Kawamura K：“先天性巨结肠症的外在神经纤维中神经细胞粘附分子 L1 的表达受损。”

Ohyama K,et al.: "Coordinat expression of L1 and 6B4 proteoglycan/phosphacan correlated with the migration of mesencephalic dopaminergic neurons in mice." Dev.Brain Res.(in press). (1998)

Ohyama K 等人：“L1 和 6B4 蛋白聚糖/磷酸聚糖的协调表达与小鼠中脑多巴胺能神经元的迁移相关。”

Kawano H,et al.: "Regenerating axons of the paraventriculo-neurohypophysial tract invaded the scar tissue that expresses extracellular matrix molecules." J.Brain Sci.(in press). (1998)

Kawano H 等人：“室旁神经垂体束的再生轴突侵入了表达细胞外基质分子的疤痕组织。”

Ohyama K,Kawano H,Kawamura K: "Localization of extracellular matrix molecules, integrins and their regulators, TGFbetas, is correlated with axon pathfinding in the spinal cord of normal and Danforth's short tail mice." Dev.Brain Res.103. 143-154 (1997)

Ohyama K、Kawano H、Kawamura K：“细胞外基质分子、整合素及其调节剂 TGFbeta 的定位与正常小鼠和丹福斯短尾小鼠脊髓中的轴突寻路相关。”

Kawano H.: "Involvement of neurocan and L1 in the formation of thalamo-cortical projection-An immunohistochemical study" Restorat.Neurol.and Nurosci.10. 167-183 (1996)

Kawano H.：“neurocan 和 L1 参与丘脑皮质投射的形成 - 一项免疫组织化学研究”Restorat.Neurol.and Nurosci.10。