Journal: Developmental biology
Article Title: Cytoskeletal polarization and cytokinetic signaling drives polar lobe formation in spiralian embryos
doi: 10.1016/j.ydbio.2019.08.020
Figure Lengend Snippet: A) Time-lapse stills of C. rubida embryos that were treated with either 0.1% DMSO or 100 μM CK666 to inhibit Arp2/3 after completing second meiosis. While the initial vegetal elongation in the zygote was observed in treated embryos (k-m), further constriction of the polar lobe neck did not occur, resulting in a shift in the position of the spindle and cleavage plane (p) and an exaggerated asymmetric cell division compared to controls. Bar, 30 μm. B) Phenotypic analysis of embryos treated with either 0.1% DMSO or 100 μM CK666, and after first division fixed and stained with Hoechst 33342 to detect DNA. Wild-type (blue bar) embryos were characterized by a larger CD blastomere and a smaller AB blastomere. The frequency of embryos with symmetric blastomeres (red bar) was not affected by CK666 treatment, whereas there was a significant increase in embryos with highly asymmetrical blastomeres (magenta bar). A small but insignificant increase in 3 cell embryos (green bar) was also observed. Errors bar represent SD for three replicate experiments per condition, >50 embryos per experiment, **p<0.01 as determined by two-way ANOVA analysis.
Article Snippet: Reagent or resource Source Identifier Antibodies Mouse monoclonal anti-Arp3 Sigma-Aldrich Cat#A5979 Rabbit anti-phospho Ser19 myosin regulatory light chain Cell signaling Cat#3671S Rat anti-αTubulin (YL1/2) Santa Cruz Biotechnology Cat#sc-53029 Rabbit anti-phosphohistone H3 Ser10 Sigma-Aldrich Cat#9701S Bacterial and Virus Strains Biological Samples Chemicals, Peptides, and Recombinant Proteins Serotonin Sigma-Aldrich Cat#H7752 CK666 Tocris Cat#3950 ZM447439 Tocris Cat#2458 Nocodazole Selleckchem Cat#S2775 Critical Commercial Assays Deposited Data Experimental Models: Cell Lines Experimental Models: Organisms/Strains Chlamys rubida San Juan Island, WA N/A Chlamys hastata San Juan Island, WA N/A Crassostrea gigas San Juan Island, WA N/A Oligonucleotides Recombinant DNA Software and Algorithms GraphPad Prism 6 Two-way ANOVA with Tukey-Kramer’s post-hoc test; GraphPad Prism v6.00 for Apple, GraphPad Software, San Diego California USA. www.graphpad.com Other Open in a separate window KEY RESOURCES TABLE Polar lobe initiation in mollusks is independent of cytokinetic signaling.
Techniques: Staining