Seki Tatsunori
Department of Anatomy, Juntendo University School of Medicine, Tokyo, Japan.
J Neurosci Res. 2002 Sep 15;69(6):772-83. doi: 10.1002/jnr.10366.
Neurons continue to be generated in the adult hippocampus. In the present study, the early developmental processes of newly generated neurons in the adult rat hippocampus were examined by confocal laser scanning microscopy using a combination of bromodeoxyuridine (BrdU) labeling and immunohistochemistry for highly polysialylated neural cell adhesion molecule (PSA-NCAM) and NeuroD, which are markers for immature neurons, and glial fibrillary acidic protein (GFAP). Rats were injected with BrdU and 2 hours, 1, 3, and 7 days after the injection, the hippocampus was processed for immunohistochemistry. One day after the injection, BrdU-labeled cells were found frequently in clusters consisting of dividing cells, putative undifferentiated cells, NeuroD-positive differentiated neurons, and GFAP-positive astrocytes. Three days later, BrdU-labeled cells were loosely aggregated and BrdU-positive fragmented nuclei were sometimes observed, suggesting that apoptosis occurred in the clusters. These BrdU-labeled nuclei were frequently associated in various ways with the processes of immature PSA-positive granule cells. They are positioned along PSA-positive apical and basal dendrites or surrounded by these processes. By 7 days after the injection, the number of the clusters was reduced and the BrdU-labeled cells had developed dendrites. These cell-to-cell associations support the hypothesis that the clustering and a microenvironment provided by the PSA-expressing immature neurons contribute to the early developmental events of adult neurogenesis, such as proliferation, differentiation, apoptosis, and neurophilic migration in the adult hippocampus.
成体海马中神经元仍在持续生成。在本研究中,通过共聚焦激光扫描显微镜,结合使用溴脱氧尿苷(BrdU)标记以及针对高度多唾液酸化神经细胞黏附分子(PSA-NCAM)、NeuroD(未成熟神经元的标志物)和胶质纤维酸性蛋白(GFAP)的免疫组织化学方法,对成年大鼠海马中新生成神经元的早期发育过程进行了检测。给大鼠注射BrdU,在注射后2小时、1天、3天和7天,对海马进行免疫组织化学处理。注射后1天,在由分裂细胞、假定的未分化细胞、NeuroD阳性的分化神经元和GFAP阳性的星形胶质细胞组成的簇中频繁发现BrdU标记的细胞。3天后,BrdU标记的细胞松散聚集,有时观察到BrdU阳性的碎片化细胞核,提示簇中发生了凋亡。这些BrdU标记的细胞核经常以各种方式与未成熟PSA阳性颗粒细胞的突起相关联。它们沿着PSA阳性的顶树突和基底树突定位,或者被这些突起包围。到注射后7天,簇的数量减少,BrdU标记的细胞长出了树突。这些细胞间的关联支持了这样的假说:由表达PSA的未成熟神经元提供的聚集和微环境有助于成年海马中成年神经发生的早期发育事件,如增殖、分化、凋亡和嗜神经性迁移。