Clonal analysis by distinct viral vectors identifies bona fide neural stem cells in the adult zebrafish telencephalon and characterizes their division properties and fate
Clonal analysis by distinct viral vectors identifies bona fide neural stem cells in the adult zebrafish telencephalon and characterizes their division properties and fate
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DOI：
10.1242/dev.058156
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发表时间：
2011-04-15
期刊：
影响因子：
4.6
通讯作者：
Bally-Cuif, Laure
中科院分区：
文献类型：
--
作者：
Rothenaigner, Ina;Krecsmarik, Monika;Bally-Cuif, Laure
斑马鱼成体大脑中广泛存在的神经发生是通过维持活跃的生发壁龛实现的。为了确定哪些祖细胞特性与这种广泛的神经发生潜能相关，我们建立了一种方法，允许在成年斑马鱼脑中使用GFP编码的逆转录病毒和慢病毒进行祖细胞转导和追踪。斑马鱼的端脑生发区由静止的放射状胶质前体细胞和活跃的神经母细胞组成。利用基于克隆病毒载体的分析能力，我们证明了这些前体细胞遵循不同的分裂模式和命运：神经母细胞主要经历有限的扩增阶段，随后是对称的神经源性分裂；相比之下，放射状胶质细胞能够在单个细胞水平上自我更新并产生不同类型的细胞，因此在体内表现出真正的神经干细胞(NSC)特性。我们还表明，放射状胶质细胞主要经历对称性的神经胶质细胞分裂，这放大了这个NSC池，并可能解释了其长期维持的原因。我们进一步证明，阻断Notch信号导致增殖细胞和克隆数量显著增加，但不影响克隆组成，表明Notch主要控制增殖而不是细胞命运。最后，通过长期追踪，我们展示了新生神经元在前脑成人回路中的功能整合。这些结果描述了成体祖细胞和神经发生的基本特征，并为使用基于病毒的技术在斑马鱼成体大脑中进行稳定的遗传操作和克隆分析开辟了道路。
Neurogenesis is widespread in the zebrafish adult brain through the maintenance of active germinal niches. To characterize which progenitor properties correlate with this extensive neurogenic potential, we set up a method that allows progenitor cell transduction and tracing in the adult zebrafish brain using GFP-encoding retro- and lentiviruses. The telencephalic germinal zone of the zebrafish comprises quiescent radial glial progenitors and actively dividing neuroblasts. Making use of the power of clonal viral vector-based analysis, we demonstrate that these progenitors follow different division modes and fates: neuroblasts primarily undergo a limited amplification phase followed by symmetric neurogenic divisions; by contrast, radial glia are capable at the single cell level of both self-renewing and generating different cell types, and hence exhibit bona fide neural stem cell (NSC) properties in vivo. We also show that radial glial cells predominantly undergo symmetric gliogenic divisions, which amplify this NSC pool and may account for its long-lasting maintenance. We further demonstrate that blocking Notch signaling results in a significant increase in proliferating cells and in the numbers of clones, but does not affect clone composition, demonstrating that Notch primarily controls proliferation rather than cell fate. Finally, through long-term tracing, we illustrate the functional integration of newborn neurons in forebrain adult circuitries. These results characterize fundamental aspects of adult progenitor cells and neurogenesis, and open the way to using virus-based technologies for stable genetic manipulations and clonal analyses in the zebrafish adult brain.