Glut1 expression and glucose uptake are upregulated around injury area of zebrafish hearts. (A) Immunostaining of MF20 (in red) and Glut1 (in green) in sham and injured hearts at 7 dpa. The nuclei were stained with DAPI (in blue). Framed areas were magnified in right panels. n: 5–6 hearts/sample. Scale bar: 100 μm or 50 μm as indicated. The experiment was repeated two times. (B) qRT-PCR was performed to examine the expression of slc2a1a in sham and injury (7 dpa) hearts. (C,D) Ex vivo glucose uptake assay. The ventricles were isolated from sham and injury hearts at 7 dpa. The isolated ventricles were incubated in 2-NBDG (a fluorescent derivative of glucose) solution for 1h, and treated with or without WZB117, a Glut1 inhibitor. The treated ventricles were captured in bright field (left panels) or 488 Channel (right panels) with a fluorescence microscope (C). The jelly-like tissues observed in the bright field images were injury areas. Framed areas were injury sties and were magnified in the right panels. The relative fluorescence intensity of the heart apex was analyzed with Image J (D). Each dot represents an individual heart (round dot, Sham group; frame dot, 7dpa group; triangle dot, 7dpa with WZB117 treatment group). n: 3–4 hearts/sample. The experiment was repeated three times. Statistical analysis was performed by Student’s two-tailed unpaired t test in GraphPad Prism 8. The p values were represented by n.s. and asterisks. n.s., p > 0.05; *, p < 0.05; ***, p < 0.001.
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