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Application of the exo utero surgery to the study on the development of the cerebral cortical

宫外手术在大脑皮质发育研究中的应用

基本信息

批准号：
08680825
负责人：
KAWAMURA Koki
金额：
$ 1.6万
依托单位：
KEIO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
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-08680825/
关键词：
rat cerebral cortex exo utero surgery development L1 antisense oligo DNA TAG-1 アンチセンスDNA ラット NCAM-H ニューロカン

项目摘要

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 exo utero sugery to clarify the developmental mechanisms of the cerebral cortex. We have already studied the in vivo development of the rat cerebral cortex and obtained the data which indicate that molecular interactions between neural cell adhesion molecules, L1 and TAG-1, and brain-specific chondroitin sulfate proteoglycan (CSPG), neurocan and phosphacan, are involved in the pathway formation of the cerebral cortex (Fukuda et al., 1997). In the developing rat cerebral cortex, L1 and TAG-1 immunoreactions were specifically localized on thalamocortical axons and cortical efferent axons, respectively. L1-bearing thalamocortical axons preferentially travel in the subplate of the cortical anlage where neurocan, was specifically expressed. In contrast, axons immunoreactive for TAG-1 evaded the subplate and run in the intermediate zone where neurocan expression was less prominent. In addition, TAG-1-bearing axons extensively invaded regions expressing another type of CSPG,phosphacan. Although both TAG-1 and L1 have been reported to bind both neurocan and phosphacan in vitro, various pattern of interactions between neural cell adhesion molecules and CSPGs may play important roles in the pathway formation of the forebrain. Based on these observation, we made microinjection of an enzyme which degrades chondrotin sulfate, antibody ageinst L1 or antisense oligo DNA of TAG-1 into the lateral ventricle of E13 mouse embryos using exo utero surgery, 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很大进展，但哺乳动物脑发育的机制仍不完全清楚。原因之一是难以观察和操纵在母亲子宫内发育的哺乳动物胚胎。本研究应用子宫外手术对小鼠胚胎脑进行了一些实验操作，以阐明大脑皮层的发育机制。我们已经研究了大鼠大脑皮层的体内发育，并获得了表明神经细胞粘附分子L1和TAG-1与脑特异性硫酸软骨素蛋白聚糖（CSPG）、神经聚糖和磷酸蛋白聚糖之间的分子相互作用参与大脑皮层的通路形成的数据（Fukuda et al.，1997年）。在发育中的大鼠大脑皮质，L1和TAG-1免疫反应特异性地定位于丘脑皮质轴突和皮质传出轴突，分别。L1轴承丘脑皮质轴突优先旅行在subplate的皮质原基，神经蛋白聚糖，是专门表达。与此相反，轴突TAG-1的免疫反应逃避基板和运行在中间区，神经聚糖的表达是不太突出。此外，TAG-1轴承轴突广泛侵入区域表达另一种类型的CSPG，磷酸蛋白聚糖。虽然TAG-1和L1都被报道在体外结合neurocan和phosphacan，但神经细胞粘附分子和CSPG之间的各种相互作用模式可能在前脑的通路形成中发挥重要作用。在此基础上，我们采用子宫外手术，将硫酸软骨素降解酶、抗ageinst L1抗体或TAG-1反义寡核苷酸DNA显微注射到E13小鼠胚胎的侧脑室，并进行脑组织学检查。该系统将为研究哺乳动物脑的发育机制提供有用的工具。