Notochord collagen markers col2a1a and col9a2 affected by dietary treatment. At 12 hpf, col2a1a expression was localized in the developing notochord of E+ (A, n = 5/5) and E− (B, n = 6/6) embryos. Arrow indicates dorsal region of the embryo. At 24 hpf, the notochord sheath in E+ and E− embryos showed col2a1a (C, D) and col9a2 (G, H) expression with a wavy notochord phenotype observed in both groups. The wavy notochord was more apparent when viewing dorsally (E, F, I, J) relative to lateral view (C, D, G, H). At 24 hpf, ~ 25% of E+ embryos (E, n = 4/12) showed a wavy col2a1a expression, while ~ 66% of E− embryos (F, n = 6/9) were obviously wavy; similarly a wavy col9a2 expression was observed in 33% of E+ embryos (I, n = 3/10), while ~ 60% of E− embryos had a wavy expression (J, n = 5/8). Scale bar represents 500 μm (C–J) and 100 μm (A, B); representative embryos are shown. Figure panels were generated with the BZ- × 700 microscope, processed with BZ-X Analyzer Software with image adjustments equally applied across time points in Adobe Photoshop. This figure was created with Adobe Photoshop v21.2.1.
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