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Fig. 4

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ZDB-FIG-180802-5

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Klatt Shaw et al., 2018 - Intracellular Calcium Mobilization Is Required for Sonic Hedgehog Signaling

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Fig. 4

RyR Function Is Required for Development of Shh-Dependent Neural Crest-Derived Neurons of the DRG and the ENS

(A–L) Lateral views of live 72 hpf Tg(isl2b:GFP) embryos with GFP-labeled Rohon-Beard neurons (arrow) and DRGs (arrowhead). Embryos were treated between 24 and 48 hpf with cyclopamine (cyc), azumolene (azum), ryanodine (ryan), N-acetyl cysteine (NAC), aldrithiol, thapsigargin (thaps), or tricaine at the indicated concentrations, or injected at the one-cell stage with ryr1a and ryr3 MOs. Arrowheads in (C and G) indicate the presence of small, faint DRGs.

(M) Quantification of embryos treated as in (A–L). DRGs present in somites 11–15 were counted. For each condition, the number of embryos analyzed is indicated. Comparisons are to control vehicle-treated embryos unless otherwise indicated. Data are represented as means ± SEM. ^∗p < 0.01; ^∗∗∗p < 0.0001; n.s., not significant.

(N–Q) Lateral views of live 78 hpf Tg(phox2bb:GFP) embryos with the ENS (arrow) labeled by GFP in control, cyclopamine-treated, ryr1a,3 morphant, or ryanodine-treated embryos. Black melanophores (pigment) are visible in each condition. Scale bar, 25 μm in (A).

See also Figure S4.

Expression Data

Genes:	EGFP GFP
Fish:	w37Tg zc7Tg w37Tg + MO1-ryr1a + MO1-ryr3 zc7Tg + MO1-ryr1a + MO1-ryr3
Conditions:	chemical treatment by environment: aldrithiol chemical treatment by environment: azumolene sodium chemical treatment: azumolene sodium chemical treatment by environment: Cyclopamine chemical treatment by environment: N-acetyl-L-cysteine chemical treatment by environment: ryanodine chemical treatment by environment: thapsigargin chemical treatment by environment: tricaine
Knockdown Reagents:	MO1-ryr1a MO1-ryr3
Anatomical Terms:	dorsal root ganglion enteric neuron Rohon-Beard neuron
Stage:	Protruding-mouth

Expression Detail

Antibody Labeling

Phenotype Data

Fish:	w37Tg zc7Tg w37Tg + MO1-ryr1a + MO1-ryr3 zc7Tg + MO1-ryr1a + MO1-ryr3
Conditions:	chemical treatment by environment: aldrithiol chemical treatment by environment: azumolene sodium chemical treatment: azumolene sodium chemical treatment by environment: Cyclopamine chemical treatment by environment: N-acetyl-L-cysteine chemical treatment by environment: ryanodine chemical treatment by environment: thapsigargin
Knockdown Reagents:	MO1-ryr1a MO1-ryr3
Observed In:	dorsal root ganglion enteric nervous system development
Stage:	Protruding-mouth

Phenotype Detail

Acknowledgments

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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