Characterization of early steps in muscle morphogenesis in a Drosophila primary culture system
Characterization of early steps in muscle morphogenesis in a Drosophila primary culture system
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DOI：
10.4161/fly.5.2.15031
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发表时间：
2011-04-01
期刊：
影响因子：
1.2
通讯作者：
Baylies, Mary K.
中科院分区：
文献类型：
--
作者：
Dobi, Krista C.;Metzger, Thomas;Baylies, Mary K.
果蝇胚胎的肌肉发生需要方正细胞(FCs)和融合能力成肌细胞(FCms)融合形成多核肌管。成肌细胞融合在胚胎中具有很好的特性，这一过程所需的许多因素已经被确定；然而，关于融合机制的一些问题仍然存在，在胚胎中回答是具有挑战性的。我们已经开发了一种改进的原代细胞培养方案来解决这些问题。利用该系统，我们确定了在培养中检测融合的最佳时间，并证实了已知的融合蛋白在胚胎中的表达和定位。重要的是，我们分别用药物latrunculin B和nocodazol破坏了肌动蛋白和微管网络，证实了肌动蛋白是成肌细胞融合所必需的，并首次表明微管在果蝇的这一过程中也是必需的。最后，我们展示了培养中的肌管采用并保持特定的肌肉特性。
Myogenesis in Drosophila embryos requires fusion between Founder cells (FCs) and Fusion Competent myoblasts (FCMs) to form multinucleate myotubes. Myoblast fusion is well characterized in embryos, and many factors required for this process have been identified; however, a number of questions pertaining to the mechanisms of fusion remain and are challenging to answer in the embryo. We have developed a modified primary cell culture protocol to address these questions in vitro. Using this system, we determined the optimal time for examining fusion in culture and confirmed that known fusion proteins are expressed and localized as in embryos. Importantly, we disrupted the actin and microtubule networks with the drugs latrunculin B and nocodazole, respectively, confirming that actin is required for myoblast fusion and showing for the first time that microtubules are also required for this process in Drosophila. Finally, we show that myotubes in culture adopt and maintain specific muscle identities.