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

Research output: Contribution to journalArticle

21 Citations (Scopus)

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., ... Dunne, M. J. (2000). Sulfonylurea receptor 1 and Kir6.2 expression in the novel human insulin-secreting cell line NES2Y. Diabetes, 49, 953-960.

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

In: Diabetes, Vol. 49, 2000, p. 953-960.

Research output: Contribution to journalArticle

Macfarlane, WM, O'Brien, RE, Barnes, PD, Shepherd, RM, Cosgrove, KE, Lindley, KJ, Aynsley-Green, A, James, RFL, Docherty, K & Dunne, MJ 2000, 'Sulfonylurea receptor 1 and Kir6.2 expression in the novel human insulin-secreting cell line NES2Y', Diabetes, vol. 49, pp. 953-960.
Macfarlane WM, O'Brien RE, Barnes PD, Shepherd RM, Cosgrove KE, Lindley KJ et al. Sulfonylurea receptor 1 and Kir6.2 expression in the novel human insulin-secreting cell line NES2Y. Diabetes. 2000;49:953-960.
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 . / Sulfonylurea receptor 1 and Kir6.2 expression in the novel human insulin-secreting cell line NES2Y. In: Diabetes. 2000 ; Vol. 49. pp. 953-960.
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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.",
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T1 - Sulfonylurea receptor 1 and Kir6.2 expression in the novel human insulin-secreting cell line NES2Y

AU - Macfarlane, W M

AU - O'Brien, R E

AU - Barnes, P D

AU - Shepherd, R M

AU - Cosgrove, K E

AU - Lindley, K J

AU - Aynsley-Green, A

AU - James, R F L

AU - Docherty, K

AU - Dunne, M J

PY - 2000

Y1 - 2000

N2 - 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.

AB - 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.

KW - PERSISTENT HYPERINSULINEMIC HYPOGLYCEMIA

KW - TRANSCRIPTION FACTOR IUF1

KW - PANCREATIC BETA-CELLS

KW - K-ATP CHANNELS

KW - FAMILIAL HYPERINSULINISM

KW - INFANCY

KW - NESIDIOBLASTOSIS

KW - STOICHIOMETRY

KW - MUTATIONS

KW - GENE

M3 - Article

VL - 49

SP - 953

EP - 960

JO - Diabetes

JF - Diabetes

SN - 0012-1797

ER -