PMC:3432770 / 27243-29281 JSONTXT

{"target":"https://pubannotation.org/docs/sourcedb/PMC/sourceid/3432770","sourcedb":"PMC","sourceid":"3432770","source_url":"http://www.ncbi.nlm.nih.gov/pmc/3432770","text":"Rpgrip1l localizes dishevelled to the basal body of zebrafish floor plate cells\nTo test whether dishevelled stability was affected uniformly in the embryo, or differentially in specific cell types or subcellular compartments, we performed whole-mount GFP immunofluorescence on embryos injected with Dsh-GFP mRNA. NLSLacZ mRNA, which codes for a nuclear form of β-galactosidase (βgal), was co-injected as a control of the presence of injected RNA in the cells. In control embryos at 12 s, Dsh-GFP concentrated in many cell types including the floor plate and notochord cells, in apical granules located at or close to the cilia (in 61% of the injected embryos, n = 39; Fig. 4 D). In double staining for βgal and GFP, all of the βgal-positive cells (n = 86 cells, 9 embryos) in the floor plate displayed Dsh-GFP granules at the cilium base, in the immediate vicinity of the basal body (Fig. 4, F and H). Co-injection of Dsh-GFP with Rpgrip1lMyc mRNA did not reveal colocalization between GFP and Myc, indicating that the dishevelled and Rpgrip1l proteins were enriched in adjacent structures at the cilium base (Fig. 4 I). In 95% of rpgrip1l morphants (n = 41), Dsh-GFP granules were either absent or severely reduced in number and size (Fig. 4 E) and found at the base of the cilium in only 9% of the βgal-positive cells (n = 78, 7 embryos; Fig. 4 G). In contrast, in inv morphants the cytoplasmic pool of Dsh-GFP was increased and the basal body pool was not grossly affected. In combined inv, rpgrip1l morphants the ciliary pool of Dsh-GFP was absent and the cytoplasmic pool was still present although reduced compared with inv morphants (Fig. S3, E–J). A Vangl2-GFP fusion protein showed a different behavior, with a homogenous localization at cell membranes in both controls and rpgrip1l morphants (Fig. S3, K–R). Together, these experiments strongly suggest that Rpgrip1l is required to maintain dishevelled levels in zebrafish floor plate cells and that it acts primarily but not exclusively on a pericentriolar pool of dishevelled.","divisions":[{"label":"Title","span":{"begin":0,"end":79}}],"tracks":[]}