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 L James, K Docherty, M J Dunne

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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 (K-ATP) 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 K-ATP channels in beta-cells, sulfonylurea receptor (SUR) 1 and Kir6.2, have operational K-ATP 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 K-ATP 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 K-ATP channels and K-ATP 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.

Original languageEnglish
Pages (from-to)953-960
Number of pages8
JournalDiabetes
Volume49
Publication statusPublished - 2000

Keywords

  • PERSISTENT HYPERINSULINEMIC HYPOGLYCEMIA
  • TRANSCRIPTION FACTOR IUF1
  • PANCREATIC BETA-CELLS
  • K-ATP CHANNELS
  • FAMILIAL HYPERINSULINISM
  • INFANCY
  • NESIDIOBLASTOSIS
  • STOICHIOMETRY
  • MUTATIONS
  • GENE

Cite this

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