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首页> 外文期刊>Development >Non-invasive long-term fluorescence live imaging of Tribolium castaneum embryos
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Non-invasive long-term fluorescence live imaging of Tribolium castaneum embryos

机译：Tribolium castaneum胚胎的无创长期荧光实时成像

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摘要

Insect development has contributed significantly to our understanding of metazoan development. However, most information has been obtained by analyzing a single species, the fruit fly Drosophila melanogaster. Embryonic development of the red flour beetle Tribolium castaneum differs fundamentally from that of Drosophila in aspects such as short-germ development,embryonic leg development, extensive extra-embryonic membrane formation and non-involuted head development. Although Tribolium has become the second most important insect model organism, previous live imaging attempts have addressed onlyspecific questions and no long-termlive imaging data of Tribolium embryogenesis have been available. By combining light sheet-based fluorescence microscopy with a novel mounting method, we achieved complete, continuous and non-invasive fluorescence live imaging of Tribolium embryogenesis at high spatiotemporal resolution. The embryos survived the 2-day or longer imaging process, developed into adults and produced fertile progeny. Our data document all morphogenetic processes from the rearrangement of the uniform blastoderm to the onset of regular muscular movement in the same embryo and in four orientations, contributing significantly to the understanding of Tribolium development. Furthermore, we created a comprehensive chronological table of Tribolium embryogenesis, integrating most previous work and providing a reference for future studies. Based on our observations, we provide evidence that serosa window closure and serosa opening, although deferred by more than 1 day, are linked. All our long-term imaging datasets are available as a resource for the community. Tribolium is only the second insect species, after Drosophila, for which non-invasive long-term fluorescence live imaging has been achieved.
机译：昆虫的发展为我们对后生动物的发展做出了重要贡献。但是，大多数信息是通过分析一种果蝇果蝇（Drosophila melanogaster）获得的。红色面粉甲虫Tribolium castaneum的胚胎发育在短胚发育，胚腿发育，广泛的胚外膜形成和非离体头部发育等方面与果蝇根本不同。尽管Tribolium已成为第二重要的昆虫模型生物，但以前的实时成像尝试仅解决了特定问题，尚无Tribolium胚胎发生的长期实时成像数据。通过将基于光片的荧光显微镜与新颖的安装方法相结合，我们以高时空分辨率实现了Tribolium胚胎发生的完整，连续和无创荧光实时成像。胚胎在2天或更长时间的成像过程中存活下来，发育为成年并产生了可育的后代。我们的数据记录了从统一胚盘重排到同一胚胎在四个方向上规则的肌肉运动开始的所有形态发生过程，为理解Tribolium的发展做出了重要贡献。此外，我们创建了Tribolium胚胎发生的综合年表，结合了以前的大部分工作，并为将来的研究提供参考。根据我们的观察，我们提供了证据表明，浆膜关闭和浆膜打开（尽管推迟了1天以上）是相关的。我们所有的长期成像数据集都可以作为社区资源。 Tribolium仅是果蝇之后的第二种昆虫，已经实现了无创长期荧光实时成像。

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