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Diabetologia

Erschienen in:

01.09.2010 | Article

Subplasmalemmal Ca²⁺ measurements in mouse pancreatic beta cells support the existence of an amplifying effect of glucose on insulin secretion

verfasst von: M. A. Ravier, R. Cheng-Xue, A. E. Palmer, J. C. Henquin, P. Gilon

Erschienen in: Diabetologia | Ausgabe 9/2010

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Abstract

Aims/hypothesis

Glucose-induced insulin secretion is attributed to a rise of beta cell cytosolic free [Ca²⁺] ([Ca²⁺]_c) (triggering pathway) and amplification of the action of Ca²⁺. This concept of amplification rests on observations that glucose can increase Ca²⁺-induced insulin secretion without further elevating an imposed already high [Ca²⁺]_c. However, it remains possible that this amplification results from an increase in [Ca²⁺] just under the plasma membrane ([Ca²⁺]_SM), which escaped detection by previous measurements of global [Ca²⁺]_c. This was the hypothesis that we tested here by measuring [Ca²⁺]_SM.

Methods

The genetically encoded Ca²⁺ indicators D3-cpv (untargeted) and LynD3-cpv (targeted to plasma membrane) were expressed in clusters of mouse beta cells. LynD3-cpv was also expressed in beta cells within intact islets. [Ca²⁺]_SM changes were monitored using total internal reflection fluorescence microscopy. Insulin secretion was measured in parallel.

Results

Beta cells expressing D3cpv or LynD3cpv displayed normal [Ca²⁺] changes and insulin secretion in response to glucose. Distinct [Ca²⁺]_SM fluctuations were detected during repetitive variations of KCl between 30 and 32–35 mmol/l, attesting to the adequate sensitivity of our system. When the amplifying pathway was evaluated (high KCl + diazoxide), increasing glucose from 3 to 15 mmol/l consistently lowered [Ca²⁺]_SM while stimulating insulin secretion approximately two fold. Blocking Ca²⁺ uptake by the endoplasmic reticulum largely attenuated the [Ca²⁺]_SM decrease produced by high glucose but did not unmask localised [Ca²⁺]_SM increases.

Conclusions/interpretation

Glucose can increase Ca²⁺-induced insulin secretion without causing further elevation of beta cell [Ca²⁺]_SM. The phenomenon is therefore a true amplification of the triggering action of Ca²⁺.

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Titel: Subplasmalemmal Ca2+ measurements in mouse pancreatic beta cells support the existence of an amplifying effect of glucose on insulin secretion
verfasst von: M. A. Ravier
R. Cheng-Xue
A. E. Palmer
J. C. Henquin
P. Gilon
Publikationsdatum: 01.09.2010
Verlag: Springer-Verlag
Erschienen in: Diabetologia / Ausgabe 9/2010
Print ISSN: 0012-186X
Elektronische ISSN: 1432-0428
DOI: https://doi.org/10.1007/s00125-010-1775-z

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