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Sulfonylurea receptor 1 and Kir6.2 expression in the novel human insulin-secreting cell line NES2Y.

  1. W M Macfarlane,
  2. R E O'Brien,
  3. P D Barnes,
  4. R M Shepherd,
  5. K E Cosgrove,
  6. K J Lindley,
  7. A Aynsley-Green,
  8. R F James,
  9. K Docherty and
  10. M J Dunne
  1. School of Biochemistry and Genetics, University of Newcastle upon Tyne, UK.
    Diabetes 2000 Jun; 49(6): 953-960. https://doi.org/10.2337/diabetes.49.6.953
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    Abstract

    NES2Y is a proliferating human insulin-secreting cell line that we have derived from a patient with persistent hyperinsulinemic hypoglycemia of infancy. This disease is characterized by unregulated insulin release despite profound hypoglycemia. NES2Y cells, like beta-cells isolated from the patient of origin, lack functional ATP-sensitive potassium channels (KATP) and also carry a defect in the insulin gene-regulatory transcription factor PDX1. Here, we report that the NES2Y beta-cells that are transfected with the genes encoding the components of KATP channels in beta-cells, sulfonylurea receptor (SUR) 1 and Kir6.2, have operational KATP channels and show normal intracellular Ca2+ and secretory responses to glucose. However, these cells, designated NESK beta-cells, have impaired insulin gene transcription responses to glucose. NES2Y beta-cells that are transfected with either Kir6.2 or SUR1 alone do not express functional KATP channels and have impaired intracellular free Ca2+ concentration-signaling responses to depolarization-dependent beta-cell agonists. These findings document that in NES2Y beta-cells, coexpression of both subunits is critically required for fully operational KATP channels and KATP channel-dependent signaling events. This article further characterizes the properties of the novel human beta-cell line, NES2Y, and documents the usefulness of these cells in diabetes-related research.

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    June 2000, 49(6)
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    Sulfonylurea receptor 1 and Kir6.2 expression in the novel human insulin-secreting cell line NES2Y.
    W M Macfarlane, R E O'Brien, P D Barnes, R M Shepherd, K E Cosgrove, K J Lindley, A Aynsley-Green, R F James, K Docherty, M J Dunne
    Diabetes Jun 2000, 49 (6) 953-960; DOI: 10.2337/diabetes.49.6.953

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    Sulfonylurea receptor 1 and Kir6.2 expression in the novel human insulin-secreting cell line NES2Y.
    W M Macfarlane, R E O'Brien, P D Barnes, R M Shepherd, K E Cosgrove, K J Lindley, A Aynsley-Green, R F James, K Docherty, M J Dunne
    Diabetes Jun 2000, 49 (6) 953-960; DOI: 10.2337/diabetes.49.6.953
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