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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神经胶质细胞是在围产期由脑室下区(SVZ)的祖细胞产生的。我们用GFP逆转录病毒标记新生大鼠前脑SVZ的祖细胞，并在P3切片上用时间推移视频显微镜观察它们的运动，以观察它们的迁移情况。我们发现了一小部分的祖细胞，这些祖细胞沿切向迁移到胼胝体(CC)，并穿过中线。这些突起保持了相对统一的形态：前导突向对侧延伸，但没有突起分支或偏离迁徙方向。净迁移需要前导突起的延长和核的移位，CC中的迁移细胞表现出这两种模式。我们证实在P3CC内存在无髓鞘轴突束，但未能检测到任何横跨CC的放射状定向神经胶质突起(Vimentin或GLAST免疫标记纤维)。对右侧扣带核进行DiI标记，观察其走向；标记纤维遍及CC，到达左侧扣带回和运动区。逆转录病毒标记的细胞在P28脑中的分布和最终命运被检测。在对侧大脑半球发现了一小部分标记细胞，不到1%，作为少突胶质细胞和星形胶质细胞，它们主要定植于扣带回和次级运动区的皮质和下面的白质。这一分布模式与骨痂纤维的投射方向相吻合。因此，神经胶质前体细胞可能与无髓鞘轴突一起穿过CC迁移到对侧大脑半球。