FIGURE

Fig. 4

ID

ZDB-FIG-230122-14

Publication

Gong et al., 2023 - Hyperaminoacidemia induces pancreatic α cell proliferation via synergism between the mTORC1 and CaSR-Gq signaling pathways

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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.

Expression Data

Expression Detail

Antibody Labeling

Phenotype Data

Phenotype Detail

Acknowledgments

This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Nat. Commun.