Morpholino oligonucleotide against ltb4r2a in zebrafish impairs macrophage migration during β-cell and liver inflammation.A, schematic representation of the β-cell injury model using Tg(ins:NTR)S950;Tg(mpeg:eGFP)gl22 zebrafish expressing nitroreductase (NTR) in β cells and green fluorescent protein in macrophages. Double-transgenic larvae were treated with metronidazole (MTZ) for 6 h, followed by imaging of islets. B, representative images of macrophages (green) and insulin (red) following injection of morpholino oligonucleotide (MO) and treatment with MTZ, as indicated (left panels), and quantitation of macrophage number within islets (right panel). Scale bar = 10 μm. C, schematic representation of the liver injury model using Tg(mpeg:eGFP)gl22 zebrafish expressing green fluorescent protein in macrophages. Transgenic larvae were injected with or without MO at 0 to 1 h post fertilization, then incubated in 5% egg solution in egg water as indicated. D, representative images of macrophages (green) and DNA (magenta) and quantification of macrophage number in the liver. The dotted line indicates the outline of the liver parenchyma and arrowheads indicate macrophages (green) within the liver parenchyma. Scale bar = 100 μm. Data are presented as mean ± SEM; and each data point represents an independent biological replicate from different animals. For statistics, unpaired two-tailed t-tests were used for comparison of two values, and 1-way ANOVA with Tukey post-test for more than two comparisons.
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