Journal: bioRxiv
Article Title: BMP antagonism is required for mandible outgrowth in zebrafish
doi: 10.64898/2026.03.11.711234
Figure Lengend Snippet: ( A ) Single Z-plane confocal images of Wheat Germ Agglutinin (WGA) stained skeletons at 6 dpf and 14 dpf highlighting Meckel’s cartilage. Yellow dashed boxes indicate the regions shown in higher-magnification insets. Chondrocytes in mutants appear enlarged and less well aligned compared to controls. ( B ) Representative 3D reconstructions of 6 dpf and 14 dpf control and mutant Meckel’s cartilage generated from WGA staining to assess cartilage volume. Quantification of cartilage volume at 6 dpf (n = 20 control, 20 dhomo; het, and 8 triple homo Meckel’s cartilages), and 14 dpf (n = 18 control and 24 dhomo; het Meckel’s cartilages) is shown. Cartilage volume is increased at 6 dpf but comparable by 14 dpf, despite the truncated and irregular morphology of mutant Meckel’s cartilages. ( C ) Representative single Z-plane confocal images of WGA-stained Meckel’s cartilage at 14 dpf, with cross-sectional views illustrating cartilage thickness along the dorsal–ventral and lateral–medial axes. Quantification of thickness along each axis (n = 18 control and 24 dhomo; het Meckel’s cartilages) reveals increased thickness in mutants across both dimensions. ( D ) Representative 3D reconstructions of five chondrocytes from WGA-stained Meckel’s cartilage in control and mutant zebrafish at 14 dpf. Quantification of chondrocyte volume at 6 dpf (five cells per cartilage; n = 20 control, 20 dhomo; het, and 8 triple homo Meckel’s cartilages), and 14 dpf (five cells per cartilage; n = 18 control and 24 dhomo; het Meckel’s cartilages) shows increased cell size in mutants by 6 dpf that persists through 14 dpf. ( E ) Representative images of single Z-plane confocal images of DAPI stained control and mutant zebrafish at 6 and 14 dpf. ( F ) Quantification of DAPI+ chondrocyte numbers across full confocal z-stacks of Meckel’s cartilage (single Z-plane shown for representation) indicates that total chondrocyte number is comparable between control and dhomo; het larvae at 6 dpf (n = 20 control and 20 dhomo; het cartilages) and 14 dpf (n = 18 control and 24 dhomo; het cartilages). However, triple homo (n = 8 cartilages) displayed increased chondrocyte numbers at 6 dpf. When 14 dpf analyses are restricted to distal regions of Meckel’s cartilage where chondrocytes adopt a side-by-side organization, dhomo; het mutants exhibit reduced chondrocyte numbers relative to controls. Arrows point to the anterior of the Meckel’s cartilage and arrowheads indicated the posterior of the Meckel’s cartilage. Scale bars, 100 µm. Error bars represent SEM.
Article Snippet: Meckel’s cartilage was outlined by Wheat Germ Agglutinin staining and traced across Z-stacks in Imaris that allowed 3D reconstruction by the Surface detection function and 3D rendering to measure the volume of Meckel’s cartilage.
Techniques: Staining, Control, Mutagenesis, Generated
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