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| Status |
Public on Apr 13, 2015 |
| Title |
Identification and Functional Analysis of Healing Regulators in Drosophila |
| Organism |
Drosophila melanogaster |
| Experiment type |
Expression profiling by array
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| Summary |
Wound healing is an essential homeostatic mechanism that maintains the epithelial barrier integrity after tissue damage. Although we know the main events participating in the healing of a wound, 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, but allow exploring many unanswered features of the healing response; e.g., which are the signal(s) responsible for initiating tissue remodeling? How is the sealing of the epithelia achieved? Or which are the inhibitory cues that cancel the healing machinery upon completion? Answering these and other questions demands in first place 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 to culture imaginal discs that allows live imaging and biochemical analysis and is healing-permissive. Employing this approach, we performed a comparative genome-wide profiling between those Drosophila imaginal cells actively involved in healing versus their non-engaged siblings. This lets us identify a set of potential wound-specific genes. Importantly, besides identifying and categorizing new genes, we functionally tested many of their gene products by genetic interference and overexpression in a healing assay. This non-saturated analysis defines a relevant set of new genes whose changes in expression levels are functionally significant for proper tissue repair. There is promise that our newly identified wound-healing genes will guide future work in the more complex mammalian wound response.
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| Overall design |
We developed a healing-permissive in vitro culture system for fly imaginal discs: we used one-channel microarrays for comparing healing-engaged cells (showing activation of the JNK signaling cascade) to cells not participating in healing (silent JNK activity) in wounded wing imaginal discs in culture. Employing this method, we aimed detecting the relevant genes involved in disc healing through microarray analysis. We compared cells actively involved in healing to those not involved and identified a whole set of upregulated or downregulated genes. They were annotated, clustered by expression profiles, chromosomal locations and presumptive functions. Most importantly, we functionally tested them genetically in a healing assay.
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| Contributor(s) |
Blanco E, Martin-Blanco E |
| Citation(s) |
25647511 |
| Submission date |
Oct 30, 2014 |
| Last update date |
May 04, 2018 |
| Contact name |
Enrique Blanco |
| E-mail(s) |
enrique.blanco@crg.eu
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| Phone |
+34 93 316 01 00
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| Organization name |
Center for Genomic Regulation (CRG)
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| Department |
Gene Regulation, Stem Cells and Cancer
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| Lab |
Epigenetic Events in Cancer (L. Di Croce's lab)
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| Street address |
Dr. Aiguader 88
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| City |
Barcelona |
| ZIP/Postal code |
08003 |
| Country |
Spain |
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| Platforms (1) |
| GPL1322 |
[Drosophila_2] Affymetrix Drosophila Genome 2.0 Array |
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| Samples (14)
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| GSM1534625 |
Wounded/GFP+, replicate II |
| GSM1534626 |
Wounded/GFP-, replicate 1A |
| GSM1534627 |
Wounded/GFP-, replicate 1B |
| GSM1534628 |
Wounded/GFP-, replicate I |
| GSM1534629 |
Wounded/GFP-, replicate II |
| GSM1534630 |
Non-wounded/GFP+, replicate 1 |
| GSM1534631 |
Non-wounded/GFP+, replicate 2 |
| GSM1534632 |
Non-wounded/GFP+, replicate 3 |
| GSM1534633 |
Non-wounded/GFP-, replicate 1 |
| GSM1534634 |
Non-wounded/GFP-, replicate 2 |
| GSM1534635 |
Non-wounded/GFP-, replicate 3 |
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| Relations |
| BioProject |
PRJNA265855 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE62863_RAW.tar |
46.8 Mb |
(http)(custom) |
TAR (of CEL) |
| Processed data included within Sample table |
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