Figure 2

Establishment of POMC^NTR and AgRP4.7^NTR transgenic fish and neuron ablation tests. (A) Schematic diagram of the DNA construct used to generate POMC^NTR and AgRP4.7^NTR transgenic zebrafish lines. Fluorescence images of (1) POMC^NTR and (3) AgRP4.7^NTR larval head at 5 days post-fertilization (dpf) (Scale bar: 200 µm). Confocal images of (2) pro-opiomelanocortin (POMC) neurons of POMC^NTR and (4) AgRP neurons of AgRP4.7^NTR at 5 dpf (Scale bar: 20 µm). (B) Left: Fluorescence images of (1) POMC^NTR transgenic zebrafish larvae at 5 dpf and (2) metronidazole (MTZ)-treated POMC^NTR transgenic zebrafish larvae at 5 dpf, n = 3. Scale bar: 500 µm. Right: quantification diagram of fluorescence intensity. All values are the mean ± standard error of the mean (SEM), n = 30. *** p < 0.001. (C) Whole mount in situ hybridization (WISH) assays showing the expression signals of pomc transcripts in POMC neurons in wild-type (WT) and POMC^NTR larvae at 5 dpf, n = 30. Scale bar: 100 µm. (D) Left: Fluorescence images of (1) AgRP4.7^NTR transgenic zebrafish larvae at 5 dpf and (2) MTZ-treated AgRP4.7^NTR transgenic zebrafish larvae at 5 dpf, n = 3. Scale bar: 500 µm. Right: quantification diagram of fluorescence intensity. All values reported as mean ± SEM, n = 30. ** p < 0.01. (E) WISH assay results showing the expression signals of agrp transcripts in AgRP neurons in WT and AgRP4.7^NTR larvae at 5 dpf. Scale bar: 100 µm.