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Live imaging of epidermal morphogenesis during the development of the adult abdominal epidermis of Drosophila


During larval stages of Drosophila development, the abdominal epidermis is composed of histoblasts (adult precursors) and larval epidermal cells (LECs). During metamorphosis, histoblasts proliferate and colonize the territories occupied by the LECs, which die and become engulfed by macrophages. This morphogenetic process is an excellent model for in vivo analysis of epithelial migration, cell division, cell death, patterning and differentiation. Here, we describe a protocol for time-lapse recording of the developing epidermis during metamorphosis. The protocol describes the removal of the pupal case (which acts as an opaque barrier to effective imaging) and mounting and imaging of specimens of different stages so that normal developmental processes are preserved. This method enables high-resolution studies over long time periods using fluorescent markers and confocal microscopy. The protocol requires 1 h for pupal dissection and mounting and, depending on the stages and genotypes to be analyzed, several more hours for preprocessing and aging and developmental staging of flies and pupae.

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Figure 1: Visualization of histoblasts in early and late pupae.
Figure 2: Orientation of live pupae on the imaging stage.
Figure 3: Dissection procedure for late pupae.
Figure 4: Timeline of adult Drosophila abdominal epithelia development.
Figure 5: Histoblast imaging in early pupae.
Figure 6: Histoblast imaging in late pupae.


  1. 1

    Martin, P. & Wood, W. Epithelial fusions in the embryo. Curr. Opin. Cell Biol. 14, 569–574 (2002).

    CAS  Article  PubMed  Google Scholar 

  2. 2

    Martín-Blanco, E. & Knust, E. Epithelial morphogenesis: filopodia at work. Curr. Biol. 11, R28–R31 (2001).

    Article  PubMed  Google Scholar 

  3. 3

    Ninov, N., Chiarelli, D.A. & Martín-Blanco, E. Extrinsic and intrinsic mechanisms directing epithelial cell sheet replacement during Drosophila metamorphosis. Development 134, 367–379 (2007).

    CAS  Article  PubMed  Google Scholar 

  4. 4

    Madhavan, M.M. & Madhavan, K. Morphogenesis of the epidermis of adult abdomen of Drosophila. J. Embryol. Exp. Morphol. 60, 1–31 (1980).

    CAS  PubMed  Google Scholar 

  5. 5

    Madhavan, M.M. & Madhavan, K. Analysis of histoblasts. Methods Mol. Biol. 247, 413–430 (2004).

    PubMed  Google Scholar 

  6. 6

    Pastor-Pareja, J.C., Grawe, F., Martín-Blanco, E. & García-Bellido, A. Invasive cell behavior during Drosophila imaginal disc eversion is mediated by the JNK signaling cascade. Dev. Cell 7, 387–399 (2004).

    CAS  Article  PubMed  Google Scholar 

  7. 7

    Le Borgne, R., Bellaïche, Y. & Schweisguth, F. Drosophila E-cadherin regulates the orientation of asymmetric cell division in the sensory organ lineage. Curr. Biol. 12, 95–104 (2002).

    CAS  Article  Google Scholar 

  8. 8

    Cordero, J.B., Larson, D.E., Craig, C.R., Hays, R. & Cagan, R. Dynamic decapentaplegic signaling regulates patterning and adhesion in the Drosophila pupal retina. Development 134, 1861–1871 (2007).

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  9. 9

    Lee, T. & Luo, L. Mosaic analysis with a repressible cell marker for studies of gene function in neuronal morphogenesis. Neuron 22, 451–461 (1999).

    CAS  Article  PubMed  Google Scholar 

  10. 10

    Brand, A.H. & Perrimon, N. Targeted gene expression as a means of altering cell fates and generating dominant phenotypes. Development 118, 401–415 (1993).

    CAS  Google Scholar 

  11. 11

    Goto, S. & Hayashi, S. Proximal to distal cell communication in the Drosophila leg provides a basis for an intercalary mechanism of limb patterning. Development 126, 3407–3413 (1999).

    CAS  PubMed  Google Scholar 

  12. 12

    Oda, H. & Tsukita, S. Dynamic features of adherens junctions during Drosophila embryonic epithelial morphogenesis revealed by a Dalpha-catenin-GFP fusion protein. Dev. Genes Evol. 209, 218–225 (1999).

    CAS  Article  PubMed  Google Scholar 

  13. 13

    Brand, A. GFP as a cell and developmental marker in the Drosophila nervous system. Methods Cell Biol. 58, 165–181 (1999).

    CAS  Article  PubMed  Google Scholar 

  14. 14

    Oda, H. & Tsukita, S. Real-time imaging of cell-cell adherens junctions reveals that Drosophila mesoderm invagination begins with two phases of apical constriction of cells. J. Cell Sci. 114, 493–501 (2001).

    CAS  PubMed  Google Scholar 

  15. 15

    Rebollo, E., Llamazares, S., Reina, J. & Gonzalez, C. Contribution of noncentrosomal microtubules to spindle assembly in Drosophila spermatocytes. PLoS Biol. 2, E8 (2004).

    Article  PubMed  PubMed Central  Google Scholar 

  16. 16

    Royou, A., Field, C., Sisson, J.C., Sullivan, W. & Karess, R. Reassessing the role and dynamics of nonmuscle myosin II during furrow formation in early Drosophila embryos. Mol. Biol. Cell 15, 838–850 (2004).

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  17. 17

    Brückner, K. et al. The PDGF/VEGF receptor controls blood cell survival in Drosophila. Dev. Cell 7, 73–84 (2004).

    Article  PubMed  Google Scholar 

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We thank Andrea Brand, Hiroki Oda, Roger Karess, Pernille Rorth and Elena Rebollo for fly stocks, Elena Rebollo for advice on setting the imaging acquisition, and Hector Herranz, lab members, reviewers and the NP editorial staff for help in improving the manuscript. Special thanks to Ashley Bruce for letting us use her equipment. N.N. holds an FPU studentship from the Spanish Ministery of Education, and E.M.-B. is supported by grants from the DGICYT (Spain) and the EU (WOUND STREP project).

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Correspondence to Enrique Martín-Blanco.

Supplementary information

Supplementary Video 1

Dissection of late pupae for live imaging (MOV 2064 kb)

Supplementary Video 2

Live imaging of early pupae (MOV 1579 kb)

Supplementary Video 3

Live imaging of late pupae (MOV 3660 kb)

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Ninov, N., Martín-Blanco, E. Live imaging of epidermal morphogenesis during the development of the adult abdominal epidermis of Drosophila. Nat Protoc 2, 3074–3080 (2007).

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