The UNC2881-Blast Helps Make The Entire UNC2881 Theory So Thrilling

We found no cases of internalizing cells that failed to accumulate apical NMY-2::GFP. No other cells visible to us at these stages accumulated myosin similarly, with the exception of places where we saw clear myosin accumulation at the cytokinetic rings of dividing cells, as expected (Fig.?6A�CD). To determine if the myosin accumulation seen occurred within each internalizing cell or on extensions from neighboring cells, we examined embryos expressing both NMY-2::GFP and a plasma membrane marker, PH::mCherry. We imaged near the surface of live embryos at 5-s intervals by spinning disk confocal microscopy. Myosin accumulation occurred within the internalizing cells, rather than their neighbors, in 50/50 MS descendants in 9 embryos, 5/6 AP24534 nmr D lineage cells in 2 embryos, 8/8 germ line precursor cells in 4 embryos, and 5/5 unidentified internalizing cells in 3 embryos (Fig.?6E�CH). To determine if apical myosin became activated in internalizing cells of diverse lineages, we examined a conserved marker for myosin UNC2881 activation. Myosin II complexes comprise two heavy chains, two essential light chains and two regulatory light chains. The phosphorylation of the two regulatory light chains at a conserved serine (p-rMLC) is required for the formation of bipolar myosin filaments and association with actin filaments (Somlyo and Somlyo, 2003). This residue in myosin regulatory light chain is phosphorylated http://www.selleckchem.com/products/Imatinib-Mesylate.html in endodermal precursor cells for a short period of the cell cycle, during cell internalization (Lee et al., 2006). We immunostained embryos expressing CEH-51::GFP, a marker for MS lineage fate (Broitman-Maduro et al., 2009) at the MS16 stage, when MS lineage cell internalization begins, using cell position and morphology to identify 34 cells that were internalizing in 10 fixed embryos. 28 of these cells were enriched for apical p-RMLC (Fig. 6I), indicating that activated apical myosin was associated with cell internalization in these cells. To determine if myosin function is required for internalization of diverse lineages, we used a temperature-sensitive, strong loss of function nmy-2 allele ( Liu et al., 2010). We shifted embryos to the restrictive temperature at a time during which many cells should be internalizing, after E lineage cell internalization should be complete, in 5 mutant embryos and 5 wild-type control embryos placed side-by-side in pairs on the same slides. In the wild-type embryos, 16?��?3.8 (mean?��?95% CI) cells internalized within 80?min of the temperature shift, whereas only 3?��?0.9 cells in the nmy-2 mutant embryos internalized in this period ( Fig. 7, p?