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First published online September 14, 2007
Journal of Experimental Biology 210, 3473-3483 (2007)
Published by The Company of Biologists 2007
doi: 10.1242/jeb.008862
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Interdependence of Ca2+ and proton movements in trout hepatocytes

Khaled H. Ahmed and Bernd Pelster*

Institut für Zoologie, and Center of Molecular Biosciences, Leopold Franzens Universität Innsbruck, Technikerstraße 25, A-6020 Innsbruck, Austria


Figure 1
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  		Fig. 1. Changes in [Ca2+]i of trout hepatocytes following addition of 0.5 µmol l–1 ionomycin (A) or 0.1 µmol l–1 thapsigargin (B) (at the time indicated by the arrows) in the absence and presence of Ca2+e. Data are means ± s.e.m. of 37–44 cells from four independent preparations in A and of 20–32 cells from three independent preparations in B.

 

Figure 2
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  		Fig. 2. Changes in pHi of trout hepatocytes following addition of 0.5 µmol l–1 ionomycin (A) or 0.1 µmol l–1 thapsigargin (B) (at the time indicated by the arrows) using Ca2+-containing medium, Ca2+-free medium and Ca2+-free medium following incubation of cells with the intracellular Ca2+ chelating agent BAPTA-AM (25 µmol l–1). Data are means ± s.e.m. of 19–128 cells from 3–16 independent preparations in A and of 28–39 cells from 3–4 independent preparations in B.

 

Figure 3
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  		Fig. 3. Changes in proton secretion rate of trout hepatocytes following addition (at the time indicated by the arrow) of 0.5 µmol l–1 ionomycin, 0.1 µmol l–1 thapsigargin or hypertonic medium (1.6x isosmolarity). Data are means ± s.e.m. from three independent preparations.

 

Figure 4
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  		Fig. 4. Changes in [Ca2+]i (A), pHi (B) and proton secretion rate (C) of trout hepatocytes upon exposure of cells to Ca2+-free medium containing 0.5 mmol l–1 EGTA followed by addition of 25 µmol l–1 of the intracellular Ca2+ chelating agent BAPTA-AM. The effect of exposure to 25 µmol l–1 BAPTA-AM in the presence of Ca2+e is depicted as BAPTA control in the three experiments. Data are means ± s.e.m. of 27–38 cells from 3–4 independent preparations in A and B and from three independent preparations in C.

 

Figure 5
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  		Fig. 5. Effect of exposure of trout hepatocytes to different pHe (at the time indicated by the arrow) on [Ca2+]i. Data are means ± s.e.m. of 43–60 cells from 6–7 independent preparations.

 

Figure 6
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  		Fig. 6. Changes in pHi (expressed as {Delta}pHi) (A) and in [Ca2+]i (expressed as % mean of basal values) (B) of trout hepatocytes following addition of 20 mmol l–1 NH4Cl. pHi measurements were performed using Ca2+-containing medium, Ca2+-free medium and Ca2+-free medium after incubation of cells with BAPTA-AM, while [Ca2+]i measurements were performed using Ca2+-containing and Ca2+-free media. Data are means ± s.e.m. of 45–58 cells in A and 24–38 cells in B from three independent preparations.

 

Figure 7
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  		Fig. 7. Changes in pHi (expressed as {Delta}pHi) (A) and in [Ca2+]i (expressed as % mean of basal values) (B) of trout hepatocytes following addition of 30 mmol l–1 Na-propionate. pHi measurements were performed using Ca2+-containing medium, Ca2+-free medium and Ca2+-free medium after incubation of cells with BAPTA-AM, while [Ca2+]i measurements were performed using Ca2+-containing and Ca2+-free media. Data are means ± s.e.m. of 33–51 cells from three independent preparations in A and 36–43 cells from four independent preparations in B.

 

Figure 8
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  		Fig. 8. Effect of 0.5 µmol l–1 cariporide (A) or removal of extracellular Na+ (B) on the hypertonicity-induced [Ca2+]i increase. Data are means ± s.e.m. of 35–76 cells from 3–7 independent preparations.

 




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