Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Spalt mediates an evolutionarily conserved switch to fibrillar muscle fate in insects

Abstract

Flying insects oscillate their wings at high frequencies of up to 1,000 Hz1,2 and produce large mechanical forces of 80 W per kilogram of muscle3. They utilize a pair of perpendicularly oriented indirect flight muscles that contain fibrillar, stretch-activated myofibres. In contrast, all other, more slowly contracting, insect body muscles have a tubular muscle morphology4. Here we identify the transcription factor Spalt major (Salm) as a master regulator of fibrillar flight muscle fate in Drosophila. salm is necessary and sufficient to induce fibrillar muscle fate. salm switches the entire transcriptional program from tubular to fibrillar fate by regulating the expression and splicing of key sarcomeric components specific to each muscle type. Spalt function is conserved in insects evolutionarily separated by 280 million years. We propose that Spalt proteins switch myofibres from tubular to fibrillar fate during development, a function potentially conserved in the vertebrate heart—a stretch-activated muscle sharing features with insect flight muscle.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Figure 1: salm specifies fibrillar flight muscle.
Figure 2: Salm expression is sufficient to induce fibrillar muscle fate.
Figure 3: vg functions upstream of salm.
Figure 4: Fibrillar insect flight muscle requires spalt function.

References

  1. 1

    Dudley, R. in The Biomechanics of Insect Flight (ed. Dudley, R. ) 75–158 (Princeton Univ. Press, 2000)

    Book  Google Scholar 

  2. 2

    Dickinson, M. Insect flight. Curr. Biol. 16, R309–R314 (2006)

    CAS  Article  Google Scholar 

  3. 3

    Lehmann, F. O. & Dickinson, M. H. The changes in power requirements and muscle efficiency during elevated force production in the fruit fly Drosophila melanogaster . J. Exp. Biol. 200, 1133–1143 (1997)

    CAS  PubMed  Google Scholar 

  4. 4

    Bernstein, S. I., O’Donnell, P. T. & Cripps, R. M. Molecular genetic analysis of muscle development, structure, and function in Drosophila . Int. Rev. Cytol. 143, 63–152 (1993)

    CAS  Article  Google Scholar 

  5. 5

    Dudley, R. in The Biomechanics of Insect Flight (ed. Dudley, R. ) 36–74 (Princeton Univ. Press, 2000)

    Book  Google Scholar 

  6. 6

    Schnorrer, F. et al. Systematic genetic analysis of muscle morphogenesis and function in Drosophila . Nature 464, 287–291 (2010)

    CAS  ADS  Article  Google Scholar 

  7. 7

    de Celis, J. F. & Barrio, R. Regulation and function of Spalt proteins during animal development. Int. J. Dev. Biol. 53, 1385–1398 (2009)

    CAS  Article  Google Scholar 

  8. 8

    Dutta, D., Anant, S., Ruiz-Gomez, M., Bate, M. & VijayRaghavan, K. Founder myoblasts and fibre number during adult myogenesis in Drosophila . Development 131, 3761–3772 (2004)

    CAS  Article  Google Scholar 

  9. 9

    Anant, S., Roy, S. & VijayRaghavan, K. Twist and Notch negatively regulate adult muscle differentiation in Drosophila . Development 125, 1361–1369 (1998)

    CAS  PubMed  Google Scholar 

  10. 10

    Franch-Marro, X. & Casanova, J. spalt-induced specification of distinct dorsal and ventral domains is required for Drosophila tracheal patterning. Dev. Biol. 250, 374–382 (2002)

    CAS  Article  Google Scholar 

  11. 11

    Mollereau, B. et al. Two-step process for photoreceptor formation in Drosophila . Nature 412, 911–913 (2001)

    CAS  ADS  Article  Google Scholar 

  12. 12

    Reedy, M. C., Bullard, B. & Vigoreaux, J. O. Flightin is essential for thick filament assembly and sarcomere stability in Drosophila flight muscles. J. Cell Biol. 151, 1483–1500 (2000)

    CAS  Article  Google Scholar 

  13. 13

    Qiu, F. et al. Myofilin, a protein in the thick filaments of insect muscle. J. Cell Sci. 118, 1527–1536 (2005)

    CAS  Article  Google Scholar 

  14. 14

    Stronach, B. E., Siegrist, S. E. & Beckerle, M. C. Two muscle-specific LIM proteins in Drosophila . J. Cell Biol. 134, 1179–1195 (1996)

    CAS  Article  Google Scholar 

  15. 15

    Bernard, F. et al. Control of apterous by vestigial drives indirect flight muscle development in Drosophila . Dev. Biol. 260, 391–403 (2003)

    CAS  Article  Google Scholar 

  16. 16

    Halder, G. et al. The Vestigial and Scalloped proteins act together to directly regulate wing-specific gene expression in Drosophila . Genes Dev. 12, 3900–3909 (1998)

    CAS  Article  Google Scholar 

  17. 17

    Maqbool, T. et al. Shaping leg muscles in Drosophila: role of ladybird, a conserved regulator of appendicular myogenesis. PLoS ONE 1, e122 (2006)

    ADS  Article  Google Scholar 

  18. 18

    Agianian, B. et al. A troponin switch that regulates muscle contraction by stretch instead of calcium. EMBO J. 23, 772–779 (2004)

    CAS  Article  Google Scholar 

  19. 19

    Arredondo, J. J. et al. Control of Drosophila paramyosin/miniparamyosin gene expression. Differential regulatory mechanisms for muscle-specific transcription. J. Biol. Chem. 276, 8278–8287 (2001)

    CAS  Article  Google Scholar 

  20. 20

    Patel, S. R. & Saide, J. D. Stretchin-klp, a novel Drosophila indirect flight muscle protein, has both myosin dependent and independent isoforms. J. Muscle Res. Cell Motil. 26, 213–224 (2005)

    CAS  Article  Google Scholar 

  21. 21

    Ayme-Southgate, A., Lasko, P., French, C. & Pardue, M. L. Characterization of the gene for mp20: a Drosophila muscle protein that is not found in asynchronous oscillatory flight muscle. J. Cell Biol. 108, 521–531 (1989)

    CAS  Article  Google Scholar 

  22. 22

    Nongthomba, U., Pasalodos-Sanchez, S., Clark, S., Clayton, J. D. & Sparrow, J. C. Expression and function of the Drosophila ACT88F actin isoform is not restricted to the indirect flight muscles. J. Muscle Res. Cell Motil. 22, 111–119 (2001)

    CAS  Article  Google Scholar 

  23. 23

    Savard, J. et al. Phylogenomic analysis reveals bees and wasps (Hymenoptera) at the base of the radiation of holometabolous insects. Genome Res. 16, 1334–1338 (2006)

    CAS  Article  Google Scholar 

  24. 24

    Tomoyasu, Y. & Denell, R. E. Larval RNAi in Tribolium (Coleoptera) for analyzing adult development. Dev. Genes Evol. 214, 575–578 (2004)

    CAS  Article  Google Scholar 

  25. 25

    Parrish, M. et al. Loss of the Sall3 gene leads to palate deficiency, abnormalities in cranial nerves, and perinatal lethality. Mol. Cell. Biol. 24, 7102–7112 (2004)

    CAS  Article  Google Scholar 

  26. 26

    Nishinakamura, R. et al. Murine homolog of SALL1 is essential for ureteric bud invasion in kidney development. Development 128, 3105–3115 (2001)

    CAS  PubMed  Google Scholar 

  27. 27

    Manisastry, S. M., Zaal, K. J. & Horowits, R. Myofibril assembly visualized by imaging N-RAP, α-actinin, and actin in living cardiomyocytes. Exp. Cell Res. 315, 2126–2139 (2009)

    CAS  Article  Google Scholar 

  28. 28

    Shiels, H. A. & White, E. The Frank–Starling mechanism in vertebrate cardiac myocytes. J. Exp. Biol. 211, 2005–2013 (2008)

    Article  Google Scholar 

  29. 29

    Surka, W. S., Kohlhase, J., Neunert, C. E., Schneider, D. S. & Proud, V. K. Unique family with Townes–Brocks syndrome, SALL1 mutation, and cardiac defects. Am. J. Med. Genet. 102, 250–257 (2001)

    CAS  Article  Google Scholar 

  30. 30

    Ranganayakulu, G., Schulz, R. A. & Olson, E. N. Wingless signaling induces nautilus expression in the ventral mesoderm of the Drosophila embryo. Dev. Biol. 176, 143–148 (1996)

    CAS  Article  Google Scholar 

  31. 31

    Grieder, N. C., Morata, G., Affolter, M. & Gehring, W. J. Spalt major controls the development of the notum and of wing hinge primordia of the Drosophila melanogaster wing imaginal disc. Dev. Biol. 329, 315–326 (2009)

    CAS  Article  Google Scholar 

  32. 32

    Rebeiz, M., Reeves, N. L. & Posakony, J. W. SCORE: a computational approach to the identification of cis-regulatory modules and target genes in whole-genome sequence data. Site clustering over random expectation. Proc. Natl Acad. Sci. USA 99, 9888–9893 (2002)

    CAS  ADS  Article  Google Scholar 

  33. 33

    Chen, E. H. & Olson, E. N. Antisocial, an intracellular adaptor protein, is required for myoblast fusion in Drosophila . Dev. Cell 1, 705–715 (2001)

    CAS  Article  Google Scholar 

  34. 34

    Newsome, T. P., Asling, B. & Dickson, B. J. Analysis of Drosophila photoreceptor axon guidance in eye-specific mosaics. Development 127, 851–860 (2000)

    CAS  PubMed  PubMed Central  Google Scholar 

  35. 35

    Kuhnlein, R. P. et al. spalt encodes an evolutionarily conserved zinc finger protein of novel structure which provides homeotic gene function in the head and tail region of the Drosophila embryo. EMBO J. 13, 168–179 (1994)

    CAS  Article  Google Scholar 

  36. 36

    Fernandes, J. J., Celniker, S. E. & VijayRaghavan, K. Development of the indirect flight muscle attachment sites in Drosophila: role of the PS integrins and the stripe gene. Dev. Biol. 176, 166–184 (1996)

    CAS  Article  Google Scholar 

  37. 37

    Klein, T. in Drosophila: Methods and Protocols (ed. Dahmann, C.) 253–264 (Humana Press, 2008).

Download references

Acknowledgements

We thank M. Affolter, D. Bäumer, M. Beckerle, B. Bullard, E. Chen, K. Clark, C. Desplan, K. Jagla, A. Lalouette, J. Posakony, D. Reiff, J. Saide, S. Sprecher, R. Schuh, G. Tanentzapf, J. Vigoreaux, K. VijayRaghavan, the Bloomington and the VDRC stock centres for fly stocks, antibodies and insect species. We are grateful to B. Dickson, I. Hein, M. Klingler and M. Sixt for discussions, and to R. Fässler for support and discussions. We thank A. Kaya-Copur, H. Knaut, M. Spletter and N. Vogt for critical comments on the manuscript. This work was supported by the Max-Planck-Society, a Career Development Award by the Human Frontier Science Programme to F.S., a Doc-fForte predoctoral fellowship from the Austrian Academy of Sciences to C.S., and DFG grants to M.F.

Author information

Affiliations

Authors

Contributions

C.S. performed most of the experiments, analysed the data and created most of the figures. F.S. acquired the time-lapse movies and performed western blots. J.D. and M.F conducted the Tribolium RNAi experiments, M.R. performed the microarray analysis, and N.J. and H.-U.D. were involved in the initial characterisation of the salm mutant phenotype. F.S. conceived and supervised the project and wrote the manuscript with input from C.S. and M.F.

Corresponding author

Correspondence to Frank Schnorrer.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Information

The file contains Supplementary Figures 1-8 with legends, Supplementary Table 1, legends for Supplementary Table 2 and Supplementary Movies 1-3 and Supplementary Data for the Tc’sal fragment for RNAi injections 3320bp. (PDF 9022 kb)

Supplementary Table 2

The table shows top 500 salm targets and IFM specific genes of microarray analysis (see Supplementary Information for full legend). (XLS 493 kb)

Supplementary Movie 1

The movie shows early IFM development in a wild-type pupa (see Supplementary Information for full legend). (MOV 4343 kb)

Supplementary Movie 2

The movie shows early IFM development in a UAS-salm-IR pupa (see Supplementary Information for full legend). (MOV 4592 kb)

Supplementary Movie 3

The movie shows salm expression in developing IFMs (see Supplementary Information for full legend). (MOV 7703 kb)

PowerPoint slides

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Schönbauer, C., Distler, J., Jährling, N. et al. Spalt mediates an evolutionarily conserved switch to fibrillar muscle fate in insects. Nature 479, 406–409 (2011). https://doi.org/10.1038/nature10559

Download citation

Further reading

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Quick links