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First published online April 8, 2004
Journal of Experimental Biology 207, 1665-1674 (2004)
Published by The Company of Biologists 2004
doi: 10.1242/jeb.00925
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Cadmium induces both pyruvate kinase and Na+/H+ exchanger activity through protein kinase C mediated signal transduction, in isolated digestive gland cells of Mytilus galloprovincialis (L.)

Stefanos Dailianis and Martha Kaloyianni*

Laboratory of Animal Physiology, Zoology Department, School of Biology, Faculty of Science, Aristotle University of Thessaloniki 54124, Greece



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Fig. 1. The effect of different concentrations of cadmium on intracellular pH (pHi) of isolated digestive gland cells of Mytilus galloprovincialis. Values are means ± S.D. of at least 7 independent experiments. In each experiment, the tissue of four animals was used. *Significantly different after cadmium treatment from control value at the respective time point. Values that share the same letter (a, b) are statistically significant different from each other at the same time point; Dunnet's test, P<0.01.

 


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Fig. 2. Effect of cadmium and various effectors on Na+ influx (µmol Na+ h–1 10–6 cells) in isolated digestive gland cells of Mytilus galloprovincialis. Cells were treated for 30 min either with cadmium at 50 µmol l–1 or cadmium (50 µmol l–1) + 20 nmol l–1 EIPA (a selective inhibitor of Na+/H+ exchanger), or cadmium (50 µmol l–1) + 20 nmol l–1 calphostin C (a specific inhibitor of protein kinase C), or cadmium (50 µmol l–1) + 10 µmol l–1 propranolol (a ß-adrenergic antagonist). See Materials and methods for details. Values are means ± S.D. of at least 4 independent experiments. In each experiment, the tissue of four animals was used. *Significant difference between control value and that observed after metal treatment; {dagger}significant difference between the value obtained after cadmium treatment with that obtained after cadmium + effector treatment; Dunnet's test, P<0.01.

 


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Fig. 3. The effect of cadmium (50 µmol l–1) and/or various effectors on pHi of isolated digestive gland cells of Mytilus galloprovincialis. Cells were treated with PMA, a potent activator of protein kinase C, alone (10 nmol l–1) or cadmium (50 µmol l–1) alone, or + 20 nmol l–1 calphostin C (a specific inhibitor of protein kinase C). The results are mean ± S.D. of at least 7 independent experiments. In each experiment, the tissue of four animals was used. *Significant difference between the values obtained after treatment of the cells with PMA or with cadmium and the control value at the respective time point; Dunnet's test, P<0.01.

 


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Fig. 4. Effect of cadmium alone and cadmium + Na+/H+ exchanger inhibitors on PK activity of isolated digestive gland cells of Mytilus galloprovincialis. 100% represents enzyme activity in the absence of cadmium (5.23±1.04 nmol min–1 mg–1 protein). Cells were treated for 30 min either with 50 µmol l–1 of cadmium alone, or 50 µmol l–1 cadmium + 10 µmol l–1 amiloride (an inhibitor of most plasma membrane Na+ transport systems) or + 20 nmol l–1 EIPA (a selective inhibitor of Na+/H+ exchanger). PK activity in the presence of 20 nmol l–1 EIPA alone after 30 min of incubation was found to be 5.38±0.7 nmol min–1 mg–1 protein. The results (expressed as % PK activity of control value) are mean ± S.D. of at least 7 independent experiments. In each experiment, the tissue from four animals was used. *Significant difference between the value after cadmium treatment and the control value; {dagger}significant difference between the values after cadmium treatment with those observed after cadmium + inhibitors; Dunnet's test, P<0.01.

 





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