Targeted expression of Nur77 in β cells restored β-cell number and insulin content in zebrafish. (A) Schematic representation of the Tg(Ins : Nur77; cmlc2:eGFP) transgenic line and the expression pattern. Expression of Nur77 was controlled by the zebrafish insulin promoter with cmlc2-driven eGFP used as an indication of transgene carriers. (B, C) Representative images (B) and quantification (C) of β-cell number in Tg(Ins:H2BmCherry), nur77−/−;Tg(Ins:H2BmCherry), Tg(Ins : Nur77);Tg(Ins:H2BmCherry) and nur77−/−; Tg(Ins : Nur77);Tg(Ins:H2BmCherry) zebrafish larvae. The number of larvae (n = 20~30). (D, E) Representative images (D) and quantification (E) of the insulin fluorescence intensity in β cells from Tg(gcg:eGFP), nur77−/−;(gcg:eGFP), Tg(Ins : Nur77);Tg(gcg:eGFP) and nur77−/−; Tg(Ins : Nur77);Tg(gcg:eGFP) zebrafish larvae. Tg(gcg:eGFP) was used to indicate the pancreatic α cell, and β cell are indicated with red fluorescence by immunostaining with insulin antibody (n = 12~15). (F) The total free glucose contents in Tg(gcg:eGFP), nur77−/−;(gcg:eGFP), Tg(Ins : Nur77);Tg(gcg:eGFP) and nur77−/−; Tg(Ins : Nur77);Tg(gcg:eGFP) zebrafish larvae (n = 3). ns, no significance; **p < 0.01, ***p < 0.001 by one-way ANOVA.
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