FIGURE

Fig. S3

ID
ZDB-FIG-170222-38
Publication
Heanue et al., 2016 - A Novel Zebrafish ret Heterozygous Model of Hirschsprung Disease Identifies a Functional Role for mapk10 as a Modifier of Enteric Nervous System Phenotype Severity
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Fig. S3

Proliferation, cell death and differentiation are all unaffected in rethu2846/+ larvae.

(A-C) Proliferation of ENS progenitor cells is equivalent in 48hpf WT (A) and rethu2846/+ (B) larvae, shown using GFP from SAGFF234A;UAS:GFP to label pixel area of ENS progenitors and BrdU (following 30 minute BrdU pulse) to quantify proliferating cells per 220um (1000 pixels) of the distal migratory wavefront of progenitors (p = 0.9182) (C). The area (in pixels squared) (D) and roundness (4*area/(π*major axis2)) (E) of the migratory wavefront was quantified, and significant increases in both parameters reflect the fact that the rethu2846/+ embryo wavefront is not elongated as is seen in WT (p<0.0001 for both area and roundness). (F-I) TUNEL staining on 42hpf WT (F,G) and rethu2846/+ (H,I) larvae does not show any TUNEL+ apoptotic cells (red, G,I) within the GFP+ migratory stream of ENCCs (F,H). (J-M) Differentiation was assayed in the SAGFF234A;UAS:GFP line by comparing HuC/D expression with GFP expression in 4dpf larvae (J,K) and counting differentiated serotonergic cells (5-HT+) within 5dpf larvae (L,M). While HuC/D+ cells show expected significantly decreased numbers in rethu2846/+ larvae (J, L, p = 0.0096 and p = 0.0045, respectively), the percentage of progenitor cells (GFP+Hu-) cells is not significantly altered (K, p = 0.3785), and the percentage of 5-HT+ cells is also not significantly altered (M, p = 0.2188).

Expression Data

Expression Detail
Antibody Labeling
Phenotype Data

Phenotype Detail
Acknowledgments
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