Salgado, AP, Santos, RM, Fernandes, AP, Tome, AR, Flatt, Peter and Rosario, LM (2000) Glucose-mediated Ca2+ signalling in single clonal insulin-secreting cells: evidence for a mixed model of cellular activation. INTERNATIONAL JOURNAL OF BIOCHEMISTRY AND CELL BIOLOGY, 32 (5). pp. 557-569. [Journal article]
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Using clonal insulin-secreting BRIN-BD11 cells, we have assessed whether the graded response of the whole cell population to glucose can be accounted for by a dose-dependent recruitment of individual cells, an amplification of the response of the recruited cells or both. Cytosolic free Ca2+ concentration ([Ca2+](i)) is an established index of beta-cell function. We used fura-2 microfluorescence techniques to assess the [Ca2+](i) responsiveness of single BRIN-BD11 cells to glucose and other secretagogues. Glucose (1-16.7 mM) evoked oscillatory [Ca2+](i) rises in these cells resembling those found in parental rat pancreatic beta-cells. The percentage of glucose-responsive cells was 11% at 1 mM and increased to 40-70% at 3-16.7 mM glucose, as assessed by a single-stimulation protocol. This profile was unrelated to possible differences in the cell cycle, as inferred from experiments where the cultured cells were synchronized by a double thymidine block protocol. Individual cells exhibited variable sensitivities to glucose (threshold range: 1-5 mM) and a variable dose-dependent amplification of the [Ca2+](i) responses (EC50 range: 2-10 mM), as assessed by a multiple-stimulation protocol. Glyceraldehyde and alpha-ketoisocaproic acid had glucose-like effects on [Ca2+](i). The data support a mixed model for the activation of insulin-secreting cells. Specifically, the graded secretory response of the whole cell population is likely to reflect both a recruitment of individual cells with different sensitivities to glucose and a dose-dependent amplification of the response of the recruited cells, (C) 2000 Elsevier Science Ltd. All rights reserved.
|Item Type:||Journal article|
|Faculties and Schools:||Faculty of Life and Health Sciences|
Faculty of Life and Health Sciences > School of Biomedical Sciences
|Research Institutes and Groups:||Biomedical Sciences Research Institute|
Biomedical Sciences Research Institute > Diabetes
|Deposited By:||Professor Peter Flatt|
|Deposited On:||13 Jan 2010 12:10|
|Last Modified:||11 Jun 2010 11:43|
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