tbg-1 encodes gamma-tubulin, a ubiquitous and highly conserved component of centrosomes in eukaryotic cells (Strome et al, 2001). Using smFISH we determined the localization of tbg-1 transcripts (red). tbg-1transcripts are detected within distinct foci throughout the cytoplasm during both the first (A) and second (B) mitosis. tbg-1 transcripts are not enriched at centrosomes or either blastomere. In contrast, GFP tagged TBG-1proteins (green signal; arrowheads) localize at centrosomes, as previously shown (Strome et al, 2001). Shown are projections from selected focal planes. Bar=10μm.
New Findings: The first observation of tbg-1 mRNA localization in early C. elegans embryos.
RNA probes targeting tbg-1 mRNAs (Quasar 670; red) were designed using Stellaris Probe Designer (Biosearch Technologies). smFISH was performed as described previously (Osborne-Nishimura et al., 2015; Shaffer et al., 2013). For hybridization, embryos were incubated with tbg-1 RNA probes (Quasar 670) at 39°C for four hours in the dark. Following hybridization, the embryos were washed and mounted with DAPI containing (blue) medium. To visualize centrosome-associated TBG-1 protein, we used transgenic strain that expresses GFP::TBG-1 (TH27; Hannak et al., (2002), green).
This work was done under a grant [7R15GM11016-02 to MHS] from the National Institute of General Medical Sciences, and research Excellence Fund from the Center of for Biomedical Research at Oakland University.
Reviewed ByAndy Golden
HistoryReceived: August 18, 2017
Accepted: August 23, 2017
Published: August 23, 2017