Localization of tbg-1 mRNAs and GFP::TBG-1 protein in Early C. elegans Embryos​.

Description

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 transcripts (red). transcripts are detected within distinct foci throughout the cytoplasm during both the first (A) and second (B) mitosis. 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.

Reagents

RNA probes targeting 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 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).