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Late sodium current (INaL) in pancreatic β-cells

  • Ion channels, receptors and transporters
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Abstract

Recent evidence of beneficial effects of ranolazine (RAN) in type II diabetes motivates interest in the role of the late sodium current (INaL) in glucose-stimulated insulin secretion. In the present work, we characterize INaL and its function in rat INS-1E cells and human islets cells. INaL was identified as steady-state current blocked by 10 μM RAN (IRAN) or 0.5 μM tetrodotoxin (TTX) (ITTX). Veratridine (VERA, 40 μM) was used as INaL enhancer. Baseline INaL was similar between INS-1E and human islet cells. In INS-1E cells, activated by glucose or tolbutamide, TTX or RAN hyperpolarized membrane potential (V m). VERA-induced depolarization was countered by TTX or RAN. ITTX and IRAN reversal potentials were negative to Na+ equilibrium one, but they approached it after Na+ substitution with Li+ or when K+ channels were blocked. This revealed INaL coupling with Na+-activated K+ current (IKNa); expression of IKNa channels (Slick/Slack) was confirmed by transcript analysis and Western blot. RAN or TTX blunted cytosolic Ca2+ response to depolarization. Long-term incubation in high (33 mM) glucose (CHG) constitutively enhanced INaL. VERA immediately increased glucose-stimulated insulin secretion. CHG increased glucose-independent secretion instead and abolished the secretory response to glucose. RAN or TTX countered VERA- and CHG-induced changes in insulin secretion. Our study demonstrated that (1) INaL was expressed in insulin-secreting cells and coupled to IKNa; INaL affected cytosolic Ca2+ but, unless enhanced, barely contributed to glucose-stimulated insulin secretion (GSIS); and (2) sustained hyperglycemic stress enhanced INaL, which contributed to the attending increase of glucose-independent insulin “leak” and GSIS impairment.

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Acknowledgments

This work was funded by grants from Gilead, Inc. (Fremont, CA) and Network Enabled Drug Design (NEDD from Regione Lombardia) to A. Zaza.

We thank Dr. Claes Wollheim and coworkers (University of Geneva, Switzerland) for kindly supplying the INS-1E cell line. We are also grateful to Drs. Luiz Belardinelli, Arvinder Dhalla, and Sridhar Rajamani of Gilead, Inc. for reading the manuscript and providing constructive comments and suggestions.

Ethical standards

The study was previously reviewed and approved by the Ethical and Scientific Committees of Azienda Ospedaliera-Ospedale Niguarda Ca’ Granda (Milan) and the University of Milano-Bicocca. Human islet donors, or their relatives on behalf, gave written consent for collection of organs and cells for transplant and research, following the statements by the Italian Ministry of Health (art. 23, law n. 91 of 1 April 1999).

Conflict of interest

The study has been partially funded by Gilead, Inc. (Fremont, CA). All authors declare the absence of further conflict of interest.

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Authors and Affiliations

  1. Department of Biotechnology and Biosciences, University of Milano-Bicocca, Piazza della Scienza 2, 20126, Milan, Italy

    Riccardo Rizzetto, Marcella Rocchetti, Luca Sala, Carlotta Ronchi, Alice Villa & Antonio Zaza

  2. Nephrology and Dialysis, Università Vita Salute San Raffaele, Milan, Italy

    Mara Ferrandi & Isabella Molinari

  3. Endocrinology Unit, Niguarda Hospital, Milan, Italy

    Federico Bertuzzi

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  1. Riccardo Rizzetto
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  2. Marcella Rocchetti
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Correspondence to Antonio Zaza.

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Rizzetto, R., Rocchetti, M., Sala, L. et al. Late sodium current (INaL) in pancreatic β-cells. Pflugers Arch - Eur J Physiol 467, 1757–1768 (2015). https://doi.org/10.1007/s00424-014-1613-0

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