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First published online May 18, 2006
Journal of Experimental Biology 209, 2156-2164 (2006)
Published by The Company of Biologists 2006
doi: 10.1242/jeb.02250

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Involvement of ryanodine-operated channels in tert-butylhydroperoxide-evoked Ca²⁺ mobilisation in pancreatic acinar cells

María A. Martínez-Burgos^1,2,*, María P. Granados², Antonio González², Juan A. Rosado², María D. Yago¹, Ginés M. Salido², Emilio Martínez-Victoria¹, Mariano Mañas¹ and José A. Pariente²

¹ Institute of Nutrition and Food Technology, Department of Physiology, University of Granada, C/Ramón y Cajal, 4. 18071, Granada, Spain
² Department of Physiology, Faculty of Veterinary Sciences, University of Extremadura, Cáceres, Spain

View larger version (19K): [in a new window]   Fig. 1. tBHP-evoked [Ca2+]c increase in isolated rat pancreatic acinar cells. Cells were perfused with 1 mmol l-1 tBHP in (A) normal-Ca2+ or (B) Ca2+-free (containing 1 mmol l-1 EGTA) medium. (C) Cells were perfused with 1 mmol l-1 tBHP followed by 2 mmol l-1 DTT in Ca2+-free medium. Traces are representative of 61 and 53 such cells taken from 10 and 8 different experiments, respectively. (D) Histogram represents the mean post-stimulus [Ca2+]c under different experimental conditions of 8-10 independent experiments. Values are means ± s.e.m. *P<0.05.

View larger version (15K): [in a new window]   Fig. 2. Mobilisation of Ca2+ in response to agonists in isolated rat pancreatic acinar cells pretreated with tBHP. Cells were initially perfused with 1 mmol l-1 tBHP followed by 1 nmol l-1 CCK-8 (A) or 1 µmol l-1 thapsigargin (TPS) (B) in Ca2+-free medium. Traces are representative of 32 and 26 such cells taken from 12 and 10 different experiments, respectively.

View larger version (15K): [in a new window]   Fig. 3. Effect of agonist-induced depletion of cytosolic Ca2+ stores on tBHP-evoked Ca2+ increase in pancreatic acinar cells. Cells were perfused with either 1 nmol l-1 CCK-8 (A) or 1 µmol l-1 thapsigargin (TPS) (B) in Ca2+-free medium, followed by 1 mmol l-1 tBHP. Traces are representative of 74 and 58 such cells taken from 12 and 10 different experiments, respectively. (C) Histogram of the mean post-stimulus [Ca2+]c under different experimental conditions in 5-12 independent experiments. Values are means ± s.e.m. *P<0.05.

View larger version (16K): [in a new window]   Fig. 4. Effect of depletion of Ca2+ mitochondrial pools on tBHP-evoked Ca2+ release in pancreatic acinar cells. (A) Cells were perfused with 0.5 µmol l-1 FCCP, followed by perfusion with 1 mmol l-1 tBHP, in Ca2+-free medium. (B) Cells were perfused with 1 mmol l-1 tBHP, followed by perfusion with 0.5 µmol l-1 FCCP, in Ca2+-free medium. Traces are representative of 27 and 18 such cells taken from 7 and 6 different experiments, respectively.

View larger version (14K): [in a new window]   Fig. 5. Effect of depletion of mitochondrial and non-mitochondrial intracellular Ca2+ stores on tBHP-evoked Ca2+ release. Pancreatic acinar cells were perfused with 0.5 µmol l-1 FCCP plus 1 µmol l-1 thapsigargin (TPS), followed by 1 mmol l-1 tBHP in a Ca2+-free medium. Trace is representative of 22 such cells taken from 5 experiments.

View larger version (14K): [in a new window]   Fig. 6. Changes in the cytosolic Ca2+ mobilisation of pancreatic acinar cells in response to perfusion with FCCP (0.5 µmol l-1), followed by tBHP (1 mmol l-1) in the presence of (A) 2-aminoethoxydiphenylborane (2-APB, 30 µmol l-1) or (B) ryanodine (50 µmol l-1). All experiments were performed in a Ca2+-free solution (1 mmol l-1 EGTA was added). Traces are representative of 37 and 46 cells taken from 2 and 3 different experiments, respectively. (C) Histogram of the mean post-stimulus [Ca2+]c under different experimental conditions of 2-3 independent experiments. Values are means ± s.e.m. *P<0.05.