Unraveling tissue regeneration pathways using chemical genetics.

TitleUnraveling tissue regeneration pathways using chemical genetics.
Publication TypeJournal Article
Year of Publication2007
AuthorsMathew LK, Sengupta S, Kawakami A, Andreasen EA, Löhr CV, Loynes CA, Renshaw SA, Peterson RT, Tanguay RL
JournalThe Journal of biological chemistry
Volume282
Issue48
Pagination35202-10
Date Published2007 Nov 30
ISSN0021-9258
KeywordsAnimals, Anti-Inflammatory Agents, Cell Differentiation, Cell Movement, Cell Proliferation, Dose-Response Relationship, Drug, Extremities, Genetic Techniques, Glucocorticoids, Macrophages, Male, Models, Anatomic, Models, Biological, Neutrophils, Regeneration, Signal Transduction, Wound Healing, Zebrafish
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 wound healing/blastema formation. In addition, knockdown of the glucocorticoid receptor restores regenerative capability to nonregenerative, 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, regeneration is not affected. Collectively, these results indicate that signaling from exogenous glucocorticoids impairs blastema formation and limits regenerative capacity through an acute inflammation-independent mechanism. These studies also demonstrate the feasibility of exploiting chemical genetics to define the pathways that govern vertebrate regeneration.

Alternate JournalJ. Biol. Chem.