Fig. 6
RyR Function Is Required in the Shh Ligand-Receiving Cell (A) Design of mosaic analysis experiments. Spatially separated NC progenitors and dorsal organizer (shield) are indicated. Formation of donor-derived Rohon-Beard (RB) and DRG neurons in mosaic embryos was determined at 78 hpf. (B–D) Examples of chimeric embryos indicating the differentiation of donor-labeled Tg(isl2b:GFP) tissue in unlabeled hosts. (B and C) Donor WT cells gave rise to both DRGs (arrows) and RBs (arrowheads) when they developed in either WT or ryr1a,3 morphant hosts. (D) ryr1a,3 morphant cells failed to produce DRGs in WT hosts. (E) The numbers of GFP-labeled RBs and DRGs present in each mosaic embryo is plotted. Lines representing best-fit analyses indicate that WT host donor cells gave rise to DRGs and RBs in a 2:3 ratio (black and magenta lines with slopes of 0.64 and 0.69, respectively). In contrast, donor cells from ryr1a,3 morphant embryos gave rise to RBs, but not DRGs (green line with a slope of 0.06). (F–M) Prox1 and En expression at 26 hpf in (F and G) shha RNA-injected, (H and I) MZptch2(−/−), (J and K) SmoM2 RNA-injected, and (L and M) dnPKA RNA-injected embryos. Azumolene treatment attenuated overproduction of Shh-dependent muscle cell types activated by Shh overexpression, loss of the Patched2 receptor, or overexpression of SmoM2. In contrast, azumolene failed to alter the development of muscle cells in embryos expressing dnPKA. Scale bar, 25 μm in (F). See also Figures S5 and S6. |
Reprinted from Developmental Cell, 45(4), Klatt Shaw, D., Gunther, D., Jurynec, M.J., Chagovetz, A.A., Ritchie, E., Grunwald, D.J., Intracellular Calcium Mobilization Is Required for Sonic Hedgehog Signaling, 512-525.e5, Copyright (2018) with permission from Elsevier. Full text @ Dev. Cell