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Regulators of G Protein Signaling (RGS) Function in Zebrafish Development
Author(s) -
Freisinger Christina M,
Westfall Trudi A,
Fisher Rory A,
Slusarski Diane C
Publication year - 2006
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.20.5.a870-b
Subject(s) - zebrafish , gene knockdown , biology , microbiology and biotechnology , regulator of g protein signaling , genetics , gene , gtpase activating protein , signal transduction , g protein
RGS proteins modulate the active life‐time of G proteins and have key roles in development. We use zebrafish ( danio rerio ) to characterize the developmental functions of RGS family members RGS3, RGS5 and RGS6. Zebrafish homologues of these RGS proteins were cloned and whole mount in situ determined the temporal and spatial expression patterns of each gene at distinct developmental stages. We next evaluated the role of each RGS protein in development using antisense oligonucleotide mediated gene knockdown. Developmental defects were identified by morphology, marker gene expression analyses and alteration to neuronal and vascular outgrowth in transgenic zebrafish lines. RGS3 knockdown generates cardiovascular and somite defects, RGS5 knockdown leads to a vascular defect and RGS6 knockdown leads to touch insensitivity and altered neuronal migration and projection including the trigeminal ganglion. Structural/functional analyses were performed to determine if direct binding to Gα subunits is required for RGS developmental actions. To this end, we generated site‐directed mutants of a conserved asparagine, required for RGS protein binding to Gα, and evaluated the ability of wild‐type and mutant RGS cDNAs to rescue knockdown phenotypes. Through use of RGS protein knockdown coupled with rescue experiments, this study provides new functional insights into the developmental roles of RGS proteins in vivo . Our evidence for distinct functional roles of structurally related RGS proteins in developmental processes raises intriguing questions regarding the mechanisms underlying these actions. (NIH HD045488 ‐01 and GM067881 ‐01)