Department of Neurology, University of California, San Francisco, CA 94143-0663, USA.
Dev Neurosci. 2010 Jul;32(2):101-13. doi: 10.1159/000279654. Epub 2010 May 4.
To investigate the effects of neonatal stroke on progenitor cells lining the lateral ventricles.
Intraventricular injection of replication-incompetent green fluorescent protein (GFP)-expressing lentivirus was performed in postnatal day 1 (P1) rats to specifically label radial glia/type B neural stem cells and ependymal cells of the lateral ventricle. A subset of animals was exposed to transient middle cerebral artery occlusion (MCAO) at P7, with mild or moderate injury confirmed by diffusion-weighted MRI and histology. Newborn cells were identified by GFP expression, location and expression of cell type-specific markers in the striatum, cortex and olfactory bulb using confocal microscopy and systematic random sampling.
Three weeks lentiviral GFP transduction of cells in the lateral ventricle, abundant GFP-expressing neurons and glia were identified in the rostral migratory stream, olfactory bulb and striatum as expected from labeling the subventricular zone (SVZ) type B neural stem cell lineage. Two weeks following mild or severe focal stroke at P7, no GFP-expressing neurons were detected in striatum or cortex although some single-labeled doublecortin+ immature neurons were detected in the penumbra. The densities of GFP+/ glial fibrillary acidic protein (GFAP)+ astrocytes and GFP+/O4+ oligodendrocytes were reduced in the striatum following MCAO (4.8 +/- 1.02 vs. 2.5 +/- 0.4 cells/high-power field, HPF; p = 0.005; 2.8+/- 1 vs. 0.5 +/- 0.2 cells/HPF, p = 0.008). Furthermore, there was a reduction of GFP+ cells in the olfactory bulb following MCAO (58.8 +/- 14.9 vs. 19.6 +/- 5.4 cells/HPF, p = 0.025). Finally, there was an increased percentage of GFP+/GFAP+ cells (70 vs. 50%), with a decreased proportion of GFP+/O4+ cells (14 vs. 30%) in injured animals.
Neurogenesis originating from cells of the lateral ventricle, including SVZ type B cells, is significantly reduced following neonatal stroke. Furthermore, neonatal stroke disrupts gliogenesis in the striatum, decreasing overall numbers of new glia and shifting the population towards astrocytes.
研究新生儿卒中对侧脑室衬里祖细胞的影响。
在出生后第 1 天（P1）的大鼠中进行复制缺陷型绿色荧光蛋白（GFP）表达慢病毒的脑室内注射，以特异性标记侧脑室的放射状胶质/ B 型神经干细胞和室管膜细胞。一部分动物在 P7 时暴露于短暂性大脑中动脉闭塞（MCAO）下，通过弥散加权 MRI 和组织学证实轻度或中度损伤。通过共聚焦显微镜和系统随机抽样，在纹状体、皮质和嗅球中使用 GFP 表达、细胞类型特异性标记物的位置和表达来鉴定新生细胞。
3 周的侧脑室慢病毒 GFP 转导后，在嗅球和纹状体中可观察到丰富的 GFP 表达神经元和胶质细胞，这与标记脑室下区（SVZ） B 型神经干细胞谱系的预期结果一致。在 P7 时发生轻度或重度局灶性卒中后 2 周，尽管在半影区检测到一些单标记的双皮质+未成熟神经元，但在纹状体或皮质中未检测到 GFP 表达的神经元。MCAO 后纹状体中 GFP+/胶质纤维酸性蛋白（GFAP）+星形胶质细胞和 GFP+/O4+少突胶质细胞的密度降低（4.8±1.02 比 2.5±0.4 个/高倍视野（HPF）；p=0.005；2.8±1 比 0.5±0.2 个/HPF，p=0.008）。此外，MCAO 后嗅球中的 GFP+细胞减少（58.8±14.9 比 19.6±5.4 个/HPF，p=0.025）。最后，损伤动物中 GFP+/GFAP+细胞的比例增加（70%比 50%），而 GFP+/O4+细胞的比例减少（14%比 30%）。
源自侧脑室细胞（包括 SVZ B 型细胞）的神经发生在新生儿卒中后显著减少。此外，新生儿卒中破坏纹状体中的神经发生，减少新生胶质细胞的总数，并使细胞群向星形胶质细胞转移。
