EphrinB2a and EphB4 functions in gallbladder and CBD formation. Unlike sibling controls (a), ephrinb1 mutants (b) show disrupted ducts in the EHB region. ephrinb2a mutants (c) exhibit dysmorphic gallbladder and CBD (46.1% show the full phenotype, 53.9% show milder phenotype, n = 13, N = 2). dephrinb1;ephrinb2a double mutants show additive EHB duct defect (100% n = 8, N = 1). eephb4a mutants show no apparent HPD defect (n = 4, N = 1). Schematics of immature HPD lumina show in a-e in domain-specific color code. f Quantification of ephrinb1, ephrinb2a, ephb4a and ephrinb1; ephrinb2a double mutant HPD formation defects at 60 hpf, scored for individual HPD domains (control: n = 12; ephrinb1: n = 14; ephrinb2a: n = 10; ephb4a: n = 4, ephrinb1;ephrinb2a: n = 8); see Supplementary Fig. 3 for averaged scores from two assessors. g–k Early duct morphogenesis defects cause dysmorphic HPD appearance at 5 dpf in ephrinb1, ephrinb2a and ephrinb1;ephrinb2a double mutants, while the HPD in ephb4a mutants appears control-like. (control: n = 10, N = 2; ephrinb1: n = 6, N = 1; ephrinb2a: n = 7; ephrinb1;ephrinb2a: n = 6; ephb4a: n = 2) l Area quantification shows the gallbladder is significantly smaller in ephrinb1; ephrinb2a double mutants at 5 dpf. m Quantification of CBD length at 5 dpf shows significant differences in ephrinb2a and ephrinb1;ephrinb2a mutants. n–p In contrast to controls (n), fewer EphrinB2a-positive gallbladder precursors form in ephb4a mutants at 52 hpf (o). p Quantification of liver and gallbladder progenitor numbers (control and ephb4an = 10). a-e: Scale bar = 15 μm, g–k = 20 μm. Statistical test: f = Fisher Exact, l, m, p = Student’s t-test. Error bars show SEM; *p < 0,05; **p < 0,01; ***p < 0,001. Supplementary movies 1, 2, 5–9, 11–13 show HPD phenotypes at 60 hpf and 5 dpf. Source data are provided as a Source Data file
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