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Genetics, Vol. 163, 997-1009, March 2003, Copyright © 2003

Coupled Mutagenesis Screens and Genetic Mapping in Zebrafish

John F. Rawlsa, Matthew R. Friedaa, Anthony R. McAdowa, Jason P. Grossa, Chad M. Claytona, Candy K. Heyena, and Stephen L. Johnsona
a Department of Genetics, Washington University School of Medicine, Saint Louis, Missouri 63110

Corresponding author: Stephen L. Johnson, 4566 Scott Ave., St. Louis, MO 63110., sjohnson{at}genetics.wustl.edu (E-mail)

Communicating editor: D. J. GRUNWALD

Forward genetic analysis is one of the principal advantages of the zebrafish model system. However, managing zebrafish mutant lines derived from mutagenesis screens and mapping the corresponding mutations and integrating them into the larger collection of mutations remain arduous tasks. To simplify and focus these endeavors, we developed an approach that facilitates the rapid mapping of new zebrafish mutations as they are generated through mutagenesis screens. We selected a minimal panel of 149 simple sequence length polymorphism markers for a first-pass genome scan in crosses involving C32 and SJD inbred lines. We also conducted a small chemical mutagenesis screen that identified several new mutations affecting zebrafish embryonic melanocyte development. Using our first-pass marker panel in bulked-segregant analysis, we were able to identify the genetic map positions of these mutations as they were isolated in our screen. Rapid mapping of the mutations facilitated stock management, helped direct allelism tests, and should accelerate identification of the affected genes. These results demonstrate the efficacy of coupling mutagenesis screens with genetic mapping.




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