Lineage analysis of newly generated neurons in organotypic culture of rat hippocampus
Lineage analysis of newly generated neurons in organotypic culture of rat hippocampus
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DOI：
10.1016/j.neures.2010.11.010
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发表时间：
2011-03-01
影响因子：
2.9
通讯作者：
Yawo, Hiromu
中科院分区：
文献类型：
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作者：
Yokose, Jun;Ishizuka, Toru;Yawo, Hiromu
在整个生命过程中，海马齿状颗粒细胞层的颗粒下区不断产生新的神经元。然而，新产生的神经元的谱系是未知的细节。在这里，使用逆转录病毒载体编码的绿色荧光蛋白，我们标记增殖细胞的器官型切片培养的大鼠出生后海马，并跟踪他们的后代在一个较长的时间。在接种后28天，使用针对细胞类型标志物如HuC/D（泛神经元标志物）、GFAP（星形胶质细胞标志物）、Prox 1（齿状颗粒细胞标志物）或NeuN（成熟神经元标志物）的特异性抗体对细胞的表型进行免疫化学鉴定。我们发现在HuC/D阳性谱系中，细胞大多为GFAP阴性。在HuC/D阳性谱系中，在细胞分裂后不久，表达EGFP的细胞通常不可追踪。这些细胞的有丝分裂后期分布在2 - 14天之间。对于同时表达Prox 1和NeuN的谱系，新生细胞在相似的时间内（2-10天）变得不可追踪。有人建议，新产生的神经元分化为成熟的齿状颗粒细胞的切片培养，一旦他们已经生存在这个关键的溯源期。(C)2010年爱思唯尔爱尔兰有限公司和日本神经科学学会。All rights reserved.
New neurons are continuously generated in the hippocampus at the subgranular zone of the dentate granule cell layer throughout life. However, the lineage of newly generated neurons is unknown in detail. Here, using a retrovirus vector encoding EGFP, we labeled proliferating cells in an organotypic slice culture of the postnatal hippocampus of rat, and tracked their descendents over a long period. At 28 days post-inoculation, the phenotypes of the cells were immunohistochemically identified using specific antibodies to cell-type markers such as HuC/D (pan-neuronal marker), GFAP (astrocyte marker), Prox1 (dentate granule cell marker) or NeuN (mature neuronal marker). We found that the cells were mostly GFAP-negative in the HuC/D-positive lineages. The EGFP-expressing cells were often untraceable shortly after cell division in the HuC/D-positive lineages. The postmitotic periods of these cells distributed between 2 and 14 days. For the lineages expressing both Prox1 and NeuN the newborn cells became untraceable in a similar period (2-10 days). It is suggested that the newly generated neurons differentiate to mature dentate granule cells in the slice culture once they have survived over this critical traceability period. (C) 2010 Elsevier Ireland Ltd and the Japan Neuroscience Society. All rights reserved.