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Analysis of Calcium Concentration Fluctuations in Hepatopancreatic R Cells of Marsupenaeus japonicus During the Molting Cycle

Laboratory of General and Comparative Physiology, Department of Biological and Environmental Sciences and Technologies, University of Lecce, Via Provinciale Lecce-Monteroni, 73100 Lecce, Italy

^* To whom correspondence should be addressed. E-mail: loredana.zilli{at}unile.it

In this study we examined the fluctuations of the intracellular calcium concentration in isolated hepatopancreatic R cells during the four molting stages of the prawn Marsupenaeus japonicus. In addition, we used the Fura-2-AM fluorescence technique to investigate the release of calcium from mitochondria and ATP-sensitive calcium stores (endoplasmic reticulum (ER), Golgi, and nucleus) into cytoplasm during the molting cycle. Results demonstrate that both the cytosolic free calcium concentration and the total cell calcium (free, bound to calcium-binding proteins, and stored in amorphous form) in the R cells strictly depend upon the molting cycle. Interestingly, the total cell calcium was higher (10 mmol l^–1) in postmolt than in premolt (1 mmol l^–1) and intermolt (0.3 mmol l^–1). The calcium released from mitochondria was higher during premolt than during postmolt and intermolt, but the amount of calcium released from ATP-sensitive calcium stores was similar during all four stages. All together, our results suggest that the mitochondria–ATP-sensitive calcium stores system does not play a key role in calcium storage during the molting cycle but that it is involved in transcellular calcium flux. We hypothesize that lysosome or membrane-clad concretion vacuoles could represent the main site of calcium storage in hepatopancreatic R cells.

Abbreviations: Cell types: B, blister-like • E, embryonic • F, fibrillar • R, resorptive