Methods to Study Roles of β-Arrestins in the Regulation of Pancreatic β-Cell Function

Stéphane Dalle (1) , Safia Costes (1) , Gyslaine Bertrand (1) , Magalie A Ravier (1)

1 IGF - Institut de Génomique Fonctionnelle

Novel findings reveal important functional roles for β-arrestin 1 and β-arrestin 2 in the regulation of insulinsecretion, β-cell survival, and β-cell mass plasticity not only by glucose but also by G-protein-coupledreceptors, such as the glucagon-like peptide-1 (GLP-1) and the pituitary adenylate cyclase-activatingpolypeptide (PACAP) receptors or GPR40, or tyrosine kinase receptors, such as the insulin receptor.Here, we describe experimental protocols to knock down β-arrestins by small interference RNA, to followsubcellular localization of β-arrestins in the cytosol and nucleus of the insulinoma INS-1E rat pancreaticβ-cell line, and to analyze β-arrestin protein expression by Western blot using INS-1E cells and isolatedmouse or human pancreatic islets. We also provide details on how to genotype β-arrestin 2 knockout(Arrb2-/-) mice and to evaluate β-arrestin-mediated roles in β-cell mass plasticity and β-cell signaling usingimmunocytochemistry on pancreatic sections or on primary dispersed β-cells from wild-type mice andArrb2-/- mice.

Mots clés: Immunofluorescence; Pancreatic beta-cell; Immunocytochemistry; Genotyping; beta-Arrestins; Small interference RNA; Western blot; Subcellular fractionation

Domaines: Biologie cellulaire

Stéphane Dalle, Safia Costes, Gyslaine Bertrand, Magalie A Ravier. Methods to Study Roles of β-Arrestins in the Regulation of Pancreatic β-Cell Function. Methods in Molecular Biology, 1957, springer; Humana Press, pp.345-364, 2019, Beta-arrestins, 978-1-4939-9157-0. ⟨10.1007/978-1-4939-9158-7_22⟩. ⟨hal-02397739⟩