Live imaging of Drosophila larval neuroblasts.

Live imaging of Drosophila larval neuroblasts.
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DOI:
10.3791/51756
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发表时间:
2014-07-07
期刊:
Journal of visualized experiments : JoVE
影响因子:
--
通讯作者:
Rusan NM
Rusan NM
中科院分区:
其他
文献类型:
--
作者:
Lerit DA;Plevock KM;Rusan NM

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干细胞不对称分裂产生两个具有不同命运潜能的子代细胞:自我更新干细胞和分化细胞。考虑到它们与发育和疾病的相关性,理解支配不对称干细胞分裂的机制一直是一个强大的研究领域。由于果蝇中枢神经系统的干细胞(或称成神经细胞)在遗传上易于处理,并且大约每小时进行一轮连续的细胞分裂,因此它们是研究干细胞分裂不可或缺的模型。大约100个神经干细胞位于两个幼虫脑叶的表面附近,使得这个模型系统对于活体成像显微镜研究特别有用。在这项工作中,我们回顾了几种广泛用于可视化干细胞分裂的方法,我们解决了这些技术的相对优点和缺点,采用解离与完整的脑组织。我们还详细介绍了我们的简化协议,用于从三龄幼虫的活细胞成像和固定的分析应用程序外植体全脑。
Stem cells divide asymmetrically to generate two progeny cells with unequal fate potential: a self-renewing stem cell and a differentiating cell. Given their relevance to development and disease, understanding the mechanisms that govern asymmetric stem cell division has been a robust area of study. Because they are genetically tractable and undergo successive rounds of cell division about once every hour, the stem cells of the Drosophila central nervous system, or neuroblasts, are indispensable models for the study of stem cell division. About 100 neural stem cells are located near the surface of each of the two larval brain lobes, making this model system particularly useful for live imaging microscopy studies. In this work, we review several approaches widely used to visualize stem cell divisions, and we address the relative advantages and disadvantages of those techniques that employ dissociated versus intact brain tissues. We also detail our simplified protocol used to explant whole brains from third instar larvae for live cell imaging and fixed analysis applications.
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