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Systematic genetic analysis of muscle morphogenesis and function in Drosophila

Abstract

Systematic genetic approaches have provided deep insight into the molecular and cellular mechanisms that operate in simple unicellular organisms. For multicellular organisms, however, the pleiotropy of gene function has largely restricted such approaches to the study of early embryogenesis. With the availability of genome-wide transgenic RNA interference (RNAi) libraries in Drosophila1,2, it is now possible to perform a systematic genetic dissection of any cell or tissue type at any stage of the lifespan. Here we apply these methods to define the genetic basis for formation and function of the Drosophila muscle. We identify a role in muscle for 2,785 genes, many of which we assign to specific functions in the organization of muscles, myofibrils or sarcomeres. Many of these genes are phylogenetically conserved, including genes implicated in mammalian sarcomere organization and human muscle diseases.

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Figure 1: Phenotypic classification of primary screen.
Figure 2: Larval muscle screen.
Figure 3: Adult muscle screen.
Figure 4: Bioinformatic analyses.

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Acknowledgements

We thank the Vienna Drosophila RNAi Center for transgenic RNAi lines, and H. Bellen, B. Bullard, J. Saide and T. Volk for their gifts of various antibodies. This work was supported by Boehringer Ingelheim GmbH, the Max-Planck Society and a postdoctoral fellowship to F.S. from the Human Frontier Science Program. Generation of a second set of RNAi lines was supported by a grant from the European Union 7th Framework Programme. A.S. and M.N. were supported by the Austrian Ministry for Science and Research (GEN-AU Bioinformatics Integration Network).

Author Contributions F.S. designed and performed the screen and other experiments, analysed the data and wrote the manuscript with B.J.D. C.S. and C.C.H.L. analysed selected muscle phenotypes in detail. G.D., K.S., M.F. and A.A. assisted in the primary screen, and G.D. also in the initial data analysis. M.R. performed the RNA microarrays. M.N. and A.S. performed the bioinformatic analyses. K.K. led the team that generated the second RNAi hairpin lines.

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Correspondence to Frank Schnorrer or Barry J. Dickson.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Information

This file contains Supplementary Methods with references, Supplementary Figures 1-7 with legends, and legends to Supplementary Tables 1-12 (tables presented in separate files). (PDF 10048 kb)

Supplementary Table 1

Phenotypes in primary screen (Fig. 1b) listed by gene and transgenic RNAi line. (XLS 2401 kb)

Supplementary Table 2

Lethality stages in primary screen (Fig. 1c) listed by gene and transgenic RNAi line. (XLS 367 kb)

Supplementary Table 3

Phenotypes of positive control genes in the primary Mef2-GAL4 screen, listed by gene and RNAi line. (XLS 39 kb)

Supplementary Table 4

Phenotypes of negative control genes in the primary Mef2-GAL4 screen. (XLS 37 kb)

Supplementary Table 5

List of genes tested with a second generation RNAi library. (XLS 87 kb)

Supplementary Table 6

Muscle morphology defects observed in the secondary screen of larval body wall muscles (Fig. 2a), as well as a list of all transformant lines tested. (XLS 60 kb)

Supplementary Table 7

Sarcomere morphology defects observed in the secondary screen of larval body wall muscles (Fig. 2b). (XLS 42 kb)

Supplementary Table 8

Supplementary Table 8 (XLS 49 kb)

Supplementary Table 9

Myofibril morphology defects observed in the secondary screen of adult IFMs (Fig. 3b),, including a list of genes with actin blobs. (XLS 44 kb)

Supplementary Table 10

Sarcomere morphology defects observed in the secondary screen of adult IFMs (Fig. 3c). (XLS 38 kb)

Supplementary Table 11

List of genes that are predicted Mef2 target genes, expressed in muscles and positive in the Mef2 RNAi screen. (XLS 27 kb)

Supplementary Table 12

List of genes positive in the Mef2 RNAi screen and found in the late muscle cluster or to be expressed in adult muscle precursors in wing discs. (XLS 20 kb)

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Schnorrer, F., Schönbauer, C., Langer, C. et al. Systematic genetic analysis of muscle morphogenesis and function in Drosophila. Nature 464, 287–291 (2010). https://doi.org/10.1038/nature08799

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