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Investigation on migration mechanisms of glial progenitors: observations of living cells labeled by dual fluorescent molecules

胶质祖细胞迁移机制的研究：双荧光分子标记活细胞的观察

基本信息

批准号：
12680770
负责人：
KAKITA Akiyoshi
金额：
$ 2.3万
依托单位：
Niigata University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2001
项目状态：
已结题

项目摘要

The great majority of glial cells of the mammalian forebrain are generated in the perinatal period from progenitors in the subventricular zone (SVZ). We investigated the migration of progenitors from the neonatal (postnatal day 0, PO) rat forebrain SVZ by labeling them in vivo with a GFP-retrovirus, and monitoring their movements by time-lapse video microscopy in P3 slices. We identified a small number of progenitors that migrated tangentially within the corpus callosum (CC) and crossed the midline. These retained a relatively uniform morphology: the leading process was extended toward the contralateral side, but showed no process branching or turning away from the migratory direction. Net migration requires the elongation of the leading process and nuclear translocation, and the migrating cells in the CC showed both modes. We confirmed the presence of unmyelinated axon bundles within the P3 CC, but failed to detect any radially directed glial processes (vimentin- or GLAST-immunolabeled fibers) spanning through the CC. The destination of the callosal fibers was examined by applying DiI to the right cingulum; the labeled fibers ran throughout the CC and reached the left cingulate and motor areas. The distribution and final fates of the retrovirus-labeled cells were examined in P28 brains. A small proportion of the labeled cells, less than 1%, were found in the contralateral hemisphere, where, as oligodendrocytes and astrocytes, they colonized predominantly the cortex and the underlying white matter of the cingulate and secondary motor areas. The distribution pattern appears to coincide well with the projection direction of the callosal fibers. Thus, glial progenitors migrate across the CC, presumably in conjunction with unmyelinated axons, to colonize the contralateral hemisphere.

哺乳动物前脑的绝大多数胶质细胞是在围产期由室管膜下区（SVZ）的祖细胞产生的。我们研究了从新生儿（出生后第0天，PO）大鼠前脑SVZ的祖细胞的迁移，通过在体内用GFP-逆转录病毒标记它们，并通过延时视频显微镜在P3切片中监测它们的运动。我们确定了少量的祖细胞，迁移切向胼胝体（CC）内，并越过中线。这些保留了一个相对统一的形态：领先的过程是向对侧延伸，但没有显示出过程分支或转向远离迁移方向。细胞的净迁移需要前导过程的延长和核转位，而CC中的迁移细胞同时具有这两种方式。我们证实了存在无髓鞘轴突束内的P3 CC，但未能检测到任何径向定向的胶质细胞过程（波形蛋白或GLAST免疫标记的纤维）跨越CC。通过将DiI应用于右侧扣带来检查胼胝体纤维的目的地;标记的纤维贯穿CC并到达左侧扣带和运动区。在P28脑中检查逆转录病毒标记细胞的分布和最终命运。标记细胞的一小部分，不到1%，被发现在对侧半球，在那里，少突胶质细胞和星形胶质细胞，他们主要定居在皮质和底层的扣带和次级运动区的白色物质。其分布与胼胝体纤维的投射方向一致。因此，神经胶质祖细胞迁移通过CC，可能与无髓鞘轴突一起，定居在对侧半球。