Phenotype Analysis and Quantification of Proliferating Cells in the Cortical Gray Matter of the Adult Rat
Phenotype Analysis and Quantification of Proliferating Cells in the Cortical Gray Matter of the Adult Rat
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DOI：
10.1267/ahc.08037
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发表时间：
2009-01-01
影响因子：
2.4
通讯作者：
Yamada, Hisao
中科院分区：
文献类型：
--
作者：
Mori, Tetsuji;Wakabayashi, Taketoshi;Yamada, Hisao
在完整的成年哺乳动物大脑中，有两个神经起源区域：海马室管膜下区和颗粒下层。即使在这些地区之外，也确实存在少量扩散的前驱物质。许多研究表明，这些细胞中的大多数是表达NG2的少突胶质细胞前体，NG2是一种硫酸软骨素蛋白多糖，大部分残留的增殖细胞似乎是内皮细胞。然而，目前仍不清楚NG2免疫阴性的增殖前体是否存在，因为以前的研究忽略了它们可能的存在。在这项研究中，我们系统地分析了完整的成年大鼠皮质灰质中增殖细胞的表型。我们改进了我们的技术，并仔细描述了增殖细胞的特征，因为在识别和定量增殖细胞时存在几个问题：NG2表达细胞的检测取决于固定条件；血管中有残留的增殖白细胞；两种抗NG2抗体产生了不同的染色模式。此外，我们还用BrdU和Ki67两种方法对增殖细胞进行了定量。我们的结果有力地表明，在完整的成人大脑皮质灰质中，只有两种类型的增殖细胞：大多数是NG2表达的细胞，包括周细胞，其余的是内皮细胞。
In intact adult mammalian brains, there are two neurogenic regions: the subependymal zone and the subgranular layer of the hippocampus. Even outside these regions, small numbers of proliferating precursors do exist. Many studies suggest that the majority of these are oligodendrocyte precursors that express NG2, a chondroitin sulfate proteoglycan, and most of the residual proliferating cells seem to be endothelial cells. However, it is still unclear whether NG2-immunonegative proliferating precursors are present, because previous studies have neglected their possible existence. In this study, we systematically analyzed the phenotypes of the proliferating cells in the intact adult rat cortical gray matter. We improved our techniques and carefully characterized the proliferating cells, because there were several problems with identifying and quantifying the proliferating cells: the detection of NG2-expressing cells was dependent on the fixation condition; there were residual proliferating leukocytes in the blood vessels; and two anti-NG2 antibodies gave rise to different staining patterns. Moreover, we used two methods, BrdU and Ki67 immunostaining, to quantify the proliferating cells. Our results strongly suggest that in the intact adult cerebral cortical gray matter, there were only two types of proliferating cells: the majority were NG2-expressing cells, including pericytes, and the rest were endothelial cells.