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Melatonin modulates Ca2+ mobilization and amylase release in response to cholecystokinin octapeptide in mouse pancreatic acinar cells

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Abstract

In the present work, we have evaluated the effect of an acute addition of melatonin on cholecystokinin octapeptide (CCK-8)-evoked Ca2+ signals and amylase secretion in mouse pancreatic acinar cells. For this purpose, freshly isolated mouse pancreatic acinar cells were loaded with fura-2 to study intracellular free Ca2+ concentration ([Ca2+]c). Amylase release and cell viability were studied employing colorimetric methods. Our results show that CCK-8 evoked a biphasic effect on amylase secretion, finding a maximum at a concentration of 0.1 nM and a reduction of secretion at higher concentrations. Pre-incubation of cells with melatonin (1 μM–1 mM) significantly attenuated enzyme secretion in response to high concentrations of CCK-8. Stimulation of cells with 1 nM CCK-8 led to a transient increase in [Ca2+]c, followed by a decrease towards a constant level. In the presence of 1 mM melatonin, stimulation of cells with CCK-8 resulted in a smaller [Ca2+]c peak response, a faster rate of decay of [Ca2+]c and lower values for the steady state of [Ca2+]c, compared with the effect of CCK-8 alone. Melatonin also reduced the oscillatory pattern of Ca2+ mobilization evoked by a physiological concentration of CCK-8 (20 pM), and completely inhibited Ca2+ mobilization induced by 10 pM CCK-8. On the other hand, Ca2+ entry from the extracellular space was not affected in the presence of melatonin. Finally, melatonin alone did not change cell viability. We conclude that melatonin, at concentrations higher than those found in blood, might regulate exocrine pancreatic function via modulation of Ca2+ signals.

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Abbreviations

[Ca2+]c :

Cytosolic free Ca2+ concentration

CCK-8:

Cholecystokinin octapeptide

EGTA:

Ethylene glycol-bis(2-aminoethylether)-N,N,N′N′-tetraacetic acid

ER:

Endoplasmic reticulum

Fura-2/AM:

Fura-2 acetoxymethyl ester

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Acknowledgements

The authors declare that there is no conflict of interest. This work was supported by Junta de Extremadura-FEDER (GR10010). Patricia Santofimia-Castaño was granted a fellowship from Junta de Extremadura (Consejería de Economía, Comercio e Innovación) and European Social Fund. The authors would like to thank Mrs. Mercedes Gomez Blázquez for her excellent technical support.

Author contributions

Patricia Santofimia-Castaño performed data acquisition and analysis. Deborah Clea Ruy is responsible for acquisition of data. Ginés M. Salido performed data interpretation and critical revision of the manuscript. Antonio González is responsible for concept/design of the research, data analysis/interpretation, drafting and approval of the manuscript.

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Authors and Affiliations

  1. Department of Physiology (Cell Physiology Research Group), University of Extremadura, Avenida Universidad s/n, 10003, Caceres, Spain

    Patricia Santofimia-Castaño, Ginés M. Salido & Antonio González

  2. Facultade de Agronomia & Medicina Veterinaria, Universidade de Brasilia, 70900-100, Brasilia, Federal District, Brazil

    Deborah Clea Ruy

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  1. Patricia Santofimia-Castaño
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Correspondence to Antonio González.

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Santofimia-Castaño, P., Ruy, D.C., Salido, G.M. et al. Melatonin modulates Ca2+ mobilization and amylase release in response to cholecystokinin octapeptide in mouse pancreatic acinar cells. J Physiol Biochem 69, 897–908 (2013). https://doi.org/10.1007/s13105-013-0267-2

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