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In vivo RNAi in the Drosophila Follicular Epithelium: Analysis of Stem Cell Maintenance, Proliferation, and Differentiation

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RNAi and Small Regulatory RNAs in Stem Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1622))

Abstract

In vivo RNAi in Drosophila facilitates simple and rapid analysis of gene functions in a cell- or tissue-specific manner. The versatility of the UAS-GAL4 system allows to control exactly where and when during development the function of a gene is depleted. The epithelium of the ovary is a particularly good model to study in a living animal how stem cells are maintained and how their descendants proliferate and differentiate. Here I provide basic information about the publicly available reagents for in vivo RNAi, and I describe how the oogenesis system can be applied to analyze stem cells and epithelial development at a histological level. Moreover, I give helpful hints to optimize the use of the UAS-GAL4 system for RNAi induction in the follicular epithelium. Finally, I provide detailed step-by-step protocols for ovary dissection, antibody stainings, and ovary mounting for microscopic analysis.

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Acknowledgments

I thank I. Woichansky and R. Kasper for the pictures shown in Figs. 2 and 4 and N. Berns for comments. Research in the lab is supported by the Deutsche Forschungsgemeinschaft (DFG) and by the Deutsche Krebshilfe. I acknowledge support of the Core Facility Live Cell Imaging Mannheim at the CBTM (DFG INST 91027/9-1 FUGG, DFG INST 91027/10-1 FUGG).

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  1. Medical Faculty Mannheim, Department of Cell and Molecular Biology, Heidelberg University, Ludolf-Krehl-Strasse 13-17, D-68167, Mannheim, Germany

    Veit Riechmann

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Correspondence to Veit Riechmann .

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  1. Department of Biology, East Carolina University, Greenville, North Carolina, USA

    Baohong Zhang

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Riechmann, V. (2017). In vivo RNAi in the Drosophila Follicular Epithelium: Analysis of Stem Cell Maintenance, Proliferation, and Differentiation. In: Zhang, B. (eds) RNAi and Small Regulatory RNAs in Stem Cells. Methods in Molecular Biology, vol 1622. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7108-4_14

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  • DOI: https://doi.org/10.1007/978-1-4939-7108-4_14

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7106-0

  • Online ISBN: 978-1-4939-7108-4

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