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Oxygen consumption oscillates in single clonal pancreatic beta-cells (HIT)

Oxygen consumption oscillates in single clonal pancreatic beta-cells (HIT)
Oxygen consumption oscillates in single clonal pancreatic beta-cells (HIT)
Based on population studies, we have hypothesized that changes in metabolism in pancreatic beta-cells precede changes in Ca2+. It is well known from single-cell Ca2+ studies that variable oscillatory patterns in Ca2+ occur in response to glucose stimulation. The present studies, using the clonal beta-cell line HIT-T-15, were undertaken to evaluate the relationship between glucose concentration, insulin secretion, and O2 consumption and to determine the Ca2+ dependency of glucose-induced changes in O2 consumption. In population studies, an excellent correlation was found between respiration and insulin secretion, with half-maximal values at approximately 1 mmol/l glucose for both respiration and secretion. In the absence of Ca2+, glucose stimulated O2 consumption but not insulin secretion. In single clonal beta-cells, a self-referencing O2 electrode was used to assess O2 consumption. Large-amplitude oscillations were found to occur in response to stimulation by glucose and were blocked by uncoupling respiration with carbonylcyanide p-(trifluoromethoxy)phenylhydrazone (FCCP). They were also blocked and respiration totally inhibited by antimycin A, an inhibitor of complex III of the respiratory chain. Half of the cells sampled (approximately 100 total) exhibited increased oscillatory O2 consumption in response to glucose. Oscillations in O2 occurred in response to glucose even in the absence of Ca2+, and their amplitude increased further on restoration of a normal extracellular Ca2+ level. These studies indicated that oscillatory O2 consumption was not dependent on Ca2+ but that the amplitude of the O2 oscillations increased in the presence of Ca2+, possibly reflecting the additional work involved in insulin secretion and Ca2+ pumping. These studies demonstrated, for the first time, a direct correlation between O2 consumption and insulin secretion, the oscillatory nature of O2 consumption in single cells, and the feasibility of using a highly sensitive noninvasive on-line self-referencing O2 electrode to monitor single beta-cell respiration.
0012-1797
1511-1516
Porterfield, D. Marshall
ad367ffb-cd69-4c9e-ac0c-f7f048538518
Corkey, Richard F.
491e731c-fc76-43c8-ae2b-5d179295dbe6
Sanger, Richard H.
eb4dde62-3c95-4a17-ac97-aba2a2ae5baf
Tornheim, Keith
a0daec50-dc61-42b3-915e-e87fcd4cb223
Smith, Peter J.S.
003de469-9420-4f12-8f0e-8e8d76d28d6c
Corkey, Barbara E.
6f4d3d0e-815c-4eec-bc7e-e8ad37384fa8
Porterfield, D. Marshall
ad367ffb-cd69-4c9e-ac0c-f7f048538518
Corkey, Richard F.
491e731c-fc76-43c8-ae2b-5d179295dbe6
Sanger, Richard H.
eb4dde62-3c95-4a17-ac97-aba2a2ae5baf
Tornheim, Keith
a0daec50-dc61-42b3-915e-e87fcd4cb223
Smith, Peter J.S.
003de469-9420-4f12-8f0e-8e8d76d28d6c
Corkey, Barbara E.
6f4d3d0e-815c-4eec-bc7e-e8ad37384fa8

Porterfield, D. Marshall, Corkey, Richard F., Sanger, Richard H., Tornheim, Keith, Smith, Peter J.S. and Corkey, Barbara E. (2000) Oxygen consumption oscillates in single clonal pancreatic beta-cells (HIT). Diabetes, 49 (9), 1511-1516. (doi:10.2337/diabetes.49.9.1511). (PMID:10969835)

Record type: Article

Abstract

Based on population studies, we have hypothesized that changes in metabolism in pancreatic beta-cells precede changes in Ca2+. It is well known from single-cell Ca2+ studies that variable oscillatory patterns in Ca2+ occur in response to glucose stimulation. The present studies, using the clonal beta-cell line HIT-T-15, were undertaken to evaluate the relationship between glucose concentration, insulin secretion, and O2 consumption and to determine the Ca2+ dependency of glucose-induced changes in O2 consumption. In population studies, an excellent correlation was found between respiration and insulin secretion, with half-maximal values at approximately 1 mmol/l glucose for both respiration and secretion. In the absence of Ca2+, glucose stimulated O2 consumption but not insulin secretion. In single clonal beta-cells, a self-referencing O2 electrode was used to assess O2 consumption. Large-amplitude oscillations were found to occur in response to stimulation by glucose and were blocked by uncoupling respiration with carbonylcyanide p-(trifluoromethoxy)phenylhydrazone (FCCP). They were also blocked and respiration totally inhibited by antimycin A, an inhibitor of complex III of the respiratory chain. Half of the cells sampled (approximately 100 total) exhibited increased oscillatory O2 consumption in response to glucose. Oscillations in O2 occurred in response to glucose even in the absence of Ca2+, and their amplitude increased further on restoration of a normal extracellular Ca2+ level. These studies indicated that oscillatory O2 consumption was not dependent on Ca2+ but that the amplitude of the O2 oscillations increased in the presence of Ca2+, possibly reflecting the additional work involved in insulin secretion and Ca2+ pumping. These studies demonstrated, for the first time, a direct correlation between O2 consumption and insulin secretion, the oscillatory nature of O2 consumption in single cells, and the feasibility of using a highly sensitive noninvasive on-line self-referencing O2 electrode to monitor single beta-cell respiration.

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Published date: September 2000
Organisations: University of Southampton

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Local EPrints ID: 190235
URI: http://eprints.soton.ac.uk/id/eprint/190235
ISSN: 0012-1797
PURE UUID: d13468a5-c7fd-4938-a9fa-ff3c14689c49
ORCID for Peter J.S. Smith: ORCID iD orcid.org/0000-0003-4400-6853

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Date deposited: 17 Jun 2011 08:11
Last modified: 29 Oct 2019 01:42

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Author: D. Marshall Porterfield
Author: Richard F. Corkey
Author: Richard H. Sanger
Author: Keith Tornheim
Author: Barbara E. Corkey

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