Genetic variation in mouse islet Ca2+ oscillations reveals novel regulators of islet function

Insufficient insulin secretion to meet metabolic demand results in diabetes. The intracellular flux of Ca2+ into β-cells triggers insulin release. Since genetics strongly influences variation in islet secretory responses, we surveyed islet Ca2+ dynamics in eight genetically diverse mouse strains. We found high strain variation in response to four conditions: 1) 8 mM glucose; 2) 8 mM glucose plus amino acids; 3) 8 mM glucose, amino acids, plus 10nM GIP; and 4) 2 mM glucose. These stimuli interrogate β-cell function, α-cell to β-cell signaling, and incretin responses. We then correlated components of the Ca2+ waveforms to islet protein abundances in the same strains used for the Ca2+ measurements. To focus on proteins relevant to human islet function, we identified human orthologues of correlated mouse proteins that are proximal to glycemic-associated SNPs in human GWAS. Several orthologues have previously been shown to regulate insulin secretion (e.g. ABCC8, PCSK1, and GCK), supporting o..., Chemicals:
All general chemicals, amino acids, BSA, DMSO, glucose, gastric inhibitory polypeptide (GIP,G2269), cOmplete Mini EDTA-free Protease Inhibitor Cocktail Tablets (11836170001), and heat-inactivated FBS (12306C) were purchased from Sigma Aldrich. RPMI 1640 base medium (11-875-093), antibiotic–antimycotic solutions (15240112), NP-40 Alternative (492016), Fura Red Ca2+ imaging dye (F3020), DiR (D12731), and agarose (BP1356-500) were purchased from ThermoFisher. Glass-bottomed culture dishes were ordered from Mattek (P35G-0-14-C). Fura Red stocks were prepared at 5 mM concentrations in DMSO, aliquoted into light-shielded tubes, and stored at -20°C until day of use (5 μM final concentration). DiR was prepared in DMSO at 2 mg/ mL, aliquoted to light-shielded tubes, and stored at 4°C until use. All imaging solutions were prepared in a bicarbonate/HEPES-buffered imaging medium (formula in Table 1). Amino acids were prepared as 100× stock in the biocarbonate/HEPES-buffered imaging mediu..., Microsoft Excel/ OpenOffice
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TITLE: Genetic variation in mouse islet Ca2+ oscillations reveals novel regulators of islet function
CITATION: Emfinger, Christopher and Clark, Lauren et al. (2022), Genetic variation in mouse islet Ca2+ oscillations reveals novel regulators of islet function, Dryad, Dataset
AUTHORS:
Christopher H. Emfinger1#, Lauren E. Clark1#, Brian Yandell2, Kathryn L. Schueler1, Shane P. Simonett1, Donnie S. Stapleton1, Kelly A. Mitok1, Matthew J. Merrins3,4, Mark P. Keller1, Alan D. Attie\*1,2

1Department of Biochemistry, University of Wisconsin-Madison, Madison, WI, 53706, USA
2Department of Statistics, University of Wisconsin-Madison, Madison, WI, 3706, USA
3Department of Medicine, Division of Endocrinology, University of Wisconsin Madison, Madison, WI, 53705, USA
4William S. Middleton Memorial Veterans Hospital, Madison, WI 53705 USA
#These authors contributed equally to this work
\*c...