ZFIN Image: Gong et al., 2023, Fig. 4

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Figure Caption

Fig. 4

Hyperaminoacidemia-induced α cell proliferation requires CaSR cell-autonomously.

a Representative images of α cells in control, gcgr^DKO, casr−/−;gcgr^DKO and Tgα^CaSR(Tg(gcga:CASR, cryaa:tagRFP));casr−/−;gcgr^DKO larvae at 5 dpf (scale bar, 10 μm). b Quantification of α cell numbers in control, gcgr^DKO, casr−/−;gcgr^DKO and Tgα^CaSR;casr−/−;gcgr^DKO larvae at 5 dpf (data represent the means ± SD, n = 12 for each group). c Representative images of EdU staining in control, gcgr^DKO, casr−/−;gcgr^DKO and Tgα^CaSR;casr−/−;gcgr^DKO larvae at 5 dpf. EdU (Red) positive α cells are indicated by arrows (scale bar, 10 μm). d Quantification of EdU positive α cell number in control, gcgr^DKO, casr−/−;gcgr^DKO and Tgα^CaSR;casr−/−;gcgr^DKO larvae at 5 dpf (Data represent means ± SD, n = 9 for each group). e Schematic for Casr knockdown by Ad^Cre-GFP and α cell proliferation assays in the primary islets of Casr^flox/flox and WT mice (created with BioRender.com). f Representative images of cells from primary WT and Casr^flox/flox mouse islets cultured with LAA and HAA media after Ad^Cre-GFP transduction. Ki67 (red) and Cre-GFP (green) double-positive α cells are indicated by red arrows, Ki67 (red) single-positive α cells are indicated by yellow arrows, primary antibody: Anti-GFP (1:300, chicken)/anti-Glucagon (1:200, mouse)/anti-Ki67 (1:150, rabbit); secondary antibody: Alexa Fluor 488 (1:1000, goat anti-chicken)/Alexa Fluor 647 (1:1000, goat anti-mouse)/Alexa Fluor 568 (1:1,000, goat anti-rabbit) (scale bar, 150 μm). g Quantification of the percentage of Ki67 positive cells among the Cre-GFP negative α cells of WT and Casr^flox/flox mouse islets cultured with LAA and HAA media after Ad^Cre-GFP transduction (data represent the means ± SD, n = 3 for each group). h Quantification of the percentage of Ki67 positive cells among the Cre-GFP positive α cells of WT and Casr^flox/flox mouse islets cultured with LAA and HAA mediums after Ad^Cre-GFP transduction (Data represent means ± SD, n = 3 for each group). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, NS indicates no significant difference (one-way ANOVA, Tukey’s multiple comparisons test, the quantifications represent individual islet sections). Source data are provided as a ^{Source Data} file.

Acknowledgments

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