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ZFIN ID: ZDB-FIG-180802-3
Klatt Shaw et al., 2018 - Intracellular Calcium Mobilization Is Required for Sonic Hedgehog Signaling. Developmental Cell   45(4):512-525.e5 Full text @ Dev. Cell
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Condition:
Anatomical Terms:
Stage: Prim-5
PHENOTYPE:
Fish:
Condition:
Observed In:
Stage: Prim-5

Fig. 2

RyR Function Is Required for Shh-Dependent Patterning of the Neural Tube

(A) Schematic of Shh-dependent neural tube (NT) patterning. The medial floorplate (MFP), lateral floorplate (LFP), and neighboring motoneurons (mn) are indicated. The notochord (ntc) and MFP are sources of Shh, designated in blue. Approximate gene expression domains of nkx2.2b, isl1, nkx6.1, and pax3 are indicated.

(B–O) Whole-mount in situ hybridization to detect neural tube markers in vehicle-treated (0.5% DMSO) and 200 μM azumolene-treated embryos. (B and C) shha-expression in the notochord (arrowhead) and MFP (arrow) is similar in control and treated embryos. (D–O) Gene expression in the neural tube is shifted upon azumolene treatment in a manner consistent with reduced Shh signaling. (D–G) Whereas foxa and foxa2 are expressed in both the MFP and LFP in control embryos, they are expressed in fewer FP cells in azumolene-treated embryos. (H–K) Expression of nkx2.2b marking the LFP is reduced, but spon1b expression marking the MFP appears unchanged in azumolene-treated embryos. (L–O) The ventral domain of the neural tube, marked by nkx6.1, is reduced, while the dorsal domain of the neural tube, marked by pax3, is expanded in azumolene-treated embryos.

(P–S) Expression of nkx2.2a in the LFP and nkx6.1 in the ventral domain of the neural tube is diminished in MZryr1a(−/−);MZryr2a(−/−);MZryr3(−/−) compared with WT.

(B, C, L–O, R, and S) Lateral views of flat mounted embryos. (D, E, H–K, P, and Q) Dorsal views of flat-mounted embryos. Prime letters, (F), and (G) are transverse cross-sections. Scale bars, 25 μm. Interval bars indicate dorsoventral extents of gene expression domains. See also Figure S3.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
foxa TU control Prim-5 floor plate ISH
TU chemical treatment by environment: azumolene sodium Prim-5 floor plate ISH
foxa2 TU control Prim-5 floor plate ISH
TU chemical treatment by environment: azumolene sodium Prim-5 floor plate ISH
nkx2.2a TU control Prim-5 lateral floor plate ISH
ryr1az42/z42 ; ryr1bz43/z43 ; ryr3z45/z45 control Prim-5 floor plate ISH
nkx2.2b TU control Prim-5 lateral floor plate ISH
TU chemical treatment by environment: azumolene sodium Prim-5 lateral floor plate ISH
nkx6.1 TU control Prim-5 neural tube ventral region ISH
TU chemical treatment by environment: azumolene sodium Prim-5 neural tube ventral region ISH
ryr1az42/z42 ; ryr1bz43/z43 ; ryr3z45/z45 control Prim-5 neural tube ventral region ISH
pax3a TU control Prim-5 neural tube dorsal region ISH
TU chemical treatment by environment: azumolene sodium Prim-5 neural tube dorsal region ISH
shha TU control Prim-5 floor plate ISH
Prim-5 notochord ISH
TU chemical treatment by environment: azumolene sodium Prim-5 floor plate ISH
Prim-5 notochord ISH
spon1b TU control Prim-5 medial floor plate ISH
TU chemical treatment by environment: azumolene sodium Prim-5 medial floor plate ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
TU chemical treatment by environment: azumolene sodium Prim-5 floor plate foxa expression decreased amount, abnormal
Prim-5 floor plate foxa2 expression decreased amount, abnormal
Prim-5 lateral floor plate nkx2.2b expression decreased amount, abnormal
Prim-5 neural tube dorsal region pax3a expression increased amount, abnormal
Prim-5 neural tube dorsal region pax3a expression increased distribution, abnormal
Prim-5 neural tube ventral region nkx6.1 expression decreased amount, abnormal
ryr1az42/z42 ; ryr1bz43/z43 ; ryr3z45/z45 control Prim-5 floor plate nkx2.2a expression decreased amount, abnormal
Prim-5 neural tube ventral region nkx6.1 expression decreased amount, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Developmental Cell for permission to reproduce figures from this article. Please note that this material may be protected by copyright.

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