Identification and functional analysis of healing regulators in Drosophila

PLoS Genet. 2015 Feb 3;11(2):e1004965. doi: 10.1371/journal.pgen.1004965. eCollection 2015.

Abstract

Wound healing is an essential homeostatic mechanism that maintains the epithelial barrier integrity after tissue damage. Although we know the overall steps in wound healing, many of the underlying molecular mechanisms remain unclear. Genetically amenable systems, such as wound healing in Drosophila imaginal discs, do not model all aspects of the repair process. However, they do allow the less understood aspects of the healing response to be explored, e.g., which signal(s) are responsible for initiating tissue remodeling? How is sealing of the epithelia achieved? Or, what inhibitory cues cancel the healing machinery upon completion? Answering these and other questions first requires the identification and functional analysis of wound specific genes. A variety of different microarray analyses of murine and humans have identified characteristic profiles of gene expression at the wound site, however, very few functional studies in healing regulation have been carried out. We developed an experimentally controlled method that is healing-permissive and that allows live imaging and biochemical analysis of cultured imaginal discs. We performed comparative genome-wide profiling between Drosophila imaginal cells actively involved in healing versus their non-engaged siblings. Sets of potential wound-specific genes were subsequently identified. Importantly, besides identifying and categorizing new genes, we functionally tested many of their gene products by genetic interference and overexpression in healing assays. This non-saturated analysis defines a relevant set of genes whose changes in expression level are functionally significant for proper tissue repair. Amongst these we identified the TCP1 chaperonin complex as a key regulator of the actin cytoskeleton essential for the wound healing response. There is promise that our newly identified wound-healing genes will guide future work in the more complex mammalian wound healing response.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actins / genetics*
  • Actins / metabolism
  • Animals
  • Cytoskeleton / genetics*
  • Cytoskeleton / pathology
  • Drosophila melanogaster
  • Epithelium / growth & development
  • Epithelium / metabolism
  • Gene Expression Regulation
  • Genome, Insect
  • Humans
  • Imaginal Discs / growth & development
  • Imaginal Discs / metabolism*
  • Imaginal Discs / pathology
  • JNK Mitogen-Activated Protein Kinases / genetics
  • MAP Kinase Signaling System / genetics
  • Regeneration / genetics
  • Signal Transduction
  • Thorax / growth & development
  • Thorax / metabolism
  • Thorax / pathology
  • Wound Healing / genetics*

Substances

  • Actins
  • JNK Mitogen-Activated Protein Kinases

Grant support

CAF and FP were supported by the EU FP6 STREP project WOUND and ST held a Spanish FPU PhD studentship. Research in the EMB laboratory is funded by grants of the EU (FP6 STREP project WOUND), the Spanish Ministry of Economy and Competitivity (DGI and CONSOLIDER grants) and the Generalitat de Catalunya (SGR). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.