γ‐aminobutyric acid stimulates β‐cell proliferation through the <scp>mTORC1</scp> / <scp>p70S6K</scp> pathway, an effect amplified by <scp>Ly49</scp> , a novel γ‐aminobutyric acid type A receptor positive allosteric modulator

Aim To examine the mechanism of action of γ-aminobutyric acid (GABA) on β-cell proliferation and investigate if co-treatment with Ly49, a novel GABA type A receptor positive allosteric modulator (GABAA -R PAM), amplifies this effect. Methods Human or mouse islets were co-treated for 4-5 days with GABA and selected receptor or cell signalling pathway modulators. Immunofluorescence was used to determine protein co-localization, cell number or proliferation, and islet size. Osmotic minipumps were surgically implanted in mice to assess Ly49 effects on pancreatic β-cells. Results Amplification of GABAA -R signalling enhanced GABA-stimulated β-cell proliferation in cultured mouse islets. Co-treatment of GABA with an inhibitor specific for PI3K, mTORC1/2, or p70S6K, abolished GABA-stimulated β-cell proliferation in mouse and human islets. Nuclear p-AktSer473 and p-p70S6KThr421/Ser424 expression in pancreatic β-cells was increased in GABA-treated mice compared with vehicle-treated mice, an effect augmented with GABA and Ly49 co-treatment. Mice co-treated with GABA and Ly49 exhibited enhanced β-cell area and proliferation compared with GABA-treated mice. Furthermore, S961 injection (an insulin receptor antagonist) resulted in enhanced plasma insulin in GABA and Ly49 co-treated mice compared with GABA-treated mice. Importantly, GABA co-treated with Ly49 increased β-cell proliferation in human islets providing a potential application for human subjects. Conclusions We show that GABA stimulates β-cell proliferation via the PI3K/mTORC1/p70S6K pathway in both mouse and human islets. Furthermore, we show that Ly49 enhances the β-cell regenerative effects of GABA, showing potential in the intervention of diabetes.