Harris Lachlan, Zalucki Oressia, Oishi Sabrina, Burne Thomas H, Jhaveri Dhanisha J, Piper Michael
The School of Biomedical Sciences, The University of Queensland, Brisbane, Queensland, Australia.
Queensland Brain Institute, The University of Queensland, Brisbane, Australia.
Dev Dyn. 2018 Jan;247(1):194-200. doi: 10.1002/dvdy.24545. Epub 2017 Aug 2.
Type 1 adult hippocampal neural stem cells (AH-NSCs) continue to generate neurons throughout life, albeit at a very low rate. The relative quiescence of this population of cells has led to many studies investigating factors that may increase their division. Current methods of identifying dividing AH-NSCs in vivo require the identification and tracing of radial processes back to nuclei within the subgranular zone. However, caveats to this approach include the time-intensive nature of identifying AH-NSCs with such a process, as well as the fact that this approach ignores the relatively more active population of horizontally oriented AH-NSCs that also reside in the subgranular zone.
Here we describe, and then verify using Hes5::GFP mice, that labeling for the cell cycle marker Ki67 and selection against the intermediate progenitor cell marker TBR2 (Ki67 ; TBR2 nuclei) is sufficient to identify dividing horizontally and radially oriented AH-NSCs in the adult mouse hippocampus.
These findings provide a simple and accurate way to quantify dividing AH-NSCs in vivo using a morphology-independent approach that will facilitate studies into neurogenesis within the hippocampal stem cell niche of the adult brain. Developmental Dynamics 247:194-200, 2018. © 2017 Wiley Periodicals, Inc.
1型成体海马神经干细胞(AH-NSCs)在整个生命过程中持续产生神经元,尽管其产生速率非常低。这群细胞的相对静止状态引发了许多研究,旨在探究可能增加其分裂的因素。目前在体内鉴定分裂中的AH-NSCs的方法需要识别并追踪放射状突起,直至颗粒下区内的细胞核。然而,这种方法存在一些问题,包括通过此过程鉴定AH-NSCs耗时较长,以及该方法忽略了同样位于颗粒下区的相对更活跃的水平取向AH-NSCs群体。
在此我们描述了一种方法,随后利用Hes5::GFP小鼠进行验证,即对细胞周期标记物Ki67进行标记,并筛选掉中间祖细胞标记物TBR2(Ki67⁺; TBR2⁻细胞核),足以在成年小鼠海马体中鉴定出水平和放射状取向的分裂中的AH-NSCs。
这些发现提供了一种简单且准确的方法,可使用一种不依赖形态学的方法在体内定量分裂中的AH-NSCs,这将有助于对成年大脑海马干细胞生态位内神经发生的研究。《发育动力学》247:194 - 200, 2018年。© 2017威利期刊公司。