PUBLICATION

Unraveling tissue regeneration pathways using chemical genetics

Authors
Mathew, L.K., Sengupta, S., Kawakami, A., Andreasen, E.A., Löhr, C.V., Loynes, C.A., Renshaw, S.A., Peterson, R.T., and Tanguay, R.L.
ID
ZDB-PUB-070920-1
Date
2007
Source
The Journal of biological chemistry   282(48): 35202-35210 (Journal)
Registered Authors
Andreasen, Eric A., Kawakami, Atsushi, Mathew, Lijoy K., Peterson, Randall, Renshaw, Steve A., Tanguay, Robyn L.
Keywords
none
MeSH Terms
  • Animals
  • Anti-Inflammatory Agents/pharmacology
  • Cell Differentiation
  • Cell Movement
  • Cell Proliferation
  • Dose-Response Relationship, Drug
  • Extremities/embryology
  • Genetic Techniques*
  • Glucocorticoids/metabolism
  • Macrophages/cytology
  • Male
  • Models, Anatomic
  • Models, Biological
  • Neutrophils/metabolism
  • Regeneration*
  • Signal Transduction
  • Wound Healing
  • Zebrafish
PubMed
17848559 Full text @ J. Biol. Chem.
Abstract
Identifying the molecular pathways that are required for regeneration remains one of the great challenges of regenerative medicine. Although genetic mutations have been useful for identifying some molecular pathways, small molecule probes of regenerative pathways might offer some advantages, including the ability to disrupt pathway function with precise temporal control. However, a vertebrate regeneration model amenable to rapid throughput small molecule screening is not currently available. We report here the development of a zebrafish early life stage fin regeneration model and its use in screening for small molecules that modulate tissue regeneration. By screening 2000 biologically active small molecules, we identified 17 that specifically inhibited regeneration. These compounds include a cluster of glucocorticoids, and we demonstrate that transient activation of the glucocorticoid receptor is sufficient to block regeneration, but only if activation occurs during a narrow window of time during wound healing/blastema formation. We further demonstrate that glucocorticoid exposure inhibit blastema formation. In addition, knockdown of the glucocorticoid receptor restores regenerative capability to non-regenerative, glucocorticoid-exposed zebrafish. To test whether the classical anti-inflammatory action of glucocorticoids is responsible for blocking regeneration, we prevented acute inflammation following amputation by antisense repression of the Pu.1 gene. Although loss of Pu.1 prevents the inflammatory response by neutrophils and macrophages, regeneration is not affected. Collectively, these results indicate that signaling from exogenous glucocorticoids impairs blastema formation and limits regenerative capability in vertebrates through an acute inflammation-independent mechanism. These studies demonstrate the feasibility of exploiting chemical genetics to define the pathways that govern vertebrate regeneration.
Genes / Markers
Figures
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Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping