Developmental origins and homeostatic mechanisms underlying adult phenotypes: multispectral sorting of pigment cells from zebrafish and non-traditional model species

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Contact PD/PI: PARICHY, DAVID M SUMMARY OF SUPPLEMENT REQUEST The parent project focuses on elucidating mechanisms of pigment cell type and pigment pattern diversification in the fishes of the genus Danio, which includes the zebrafish D. rerio, as well as more distantly related species. This equipment supplement request seeks purchase of a Cytek Aurora CS fluorescence activated cell sorter that will greatly enhance our ability to meet the original goals of the award, which require extensive sin- gle cell RNA-sequencing analyses of pigment cells from wild-type and mutant backgrounds of zebrafish and other danios, an approach that is proving to be technically intractable and financially infeasible with the instru- ment currently available. The Cytek Aurora CS will also permit us to take an innovative, in vitro approach to elucidating mechanisms of cell-cell interactions during pattern formation in zebrafish, while uniquely allowing us to test new hypotheses pertaining to evolutionary diversification of cell types using marine fishes (e.g., the wrasse Cirrhilabrus finfenmaa) and other species (e.g., rasbora Trigonostigma heteromorpha) that cannot be bred in the laboratory and that lack the genetic and genomic resources of zebrafish. 1 Project Summary/Abstract Page 6