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First published online February 1, 2008
Journal of Experimental Biology 211, 577-586 (2008)
Published by The Company of Biologists 2008
doi: 10.1242/jeb.011262
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Effects of cadmium on cellular protein and glutathione synthesis and expression of stress proteins in eastern oysters, Crassostrea virginica Gmelin

Anna V. Ivanina*, Anton S. Cherkasov* and Inna M. Sokolova{dagger}

Biology Department, University of North Carolina at Charlotte, 9201 University City Blvd, Charlotte, NC 28223, USA


Figure 1
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  		Fig. 1. Dose-dependent accumulation of cadmium (Cd) in isolated gill and hepatopancreas (HP) cells of C. virginica. Mean Cd burdens (y-axis) accumulated after 4 h exposure in varying Cd concentrations (x-axis) at 20°C are shown. Vertical bars represent s.e.m. (N=5).

 

Figure 2
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  		Fig. 2. Effects of cadmium (Cd) exposure on protein synthesis rates in gill and hepatopancreas (HP) cells of oysters. (A, insert) Protein synthesis rates in control gill and HP cells estimated by the rate of incorporation of [3H]Leu. Vertical bars represent s.e.m. (N=8). (B) Changes in [3H]Leu incorporation in response to Cd exposure for 4 h at 20°C expressed as % of the respective controls. Vertical bars represent s.e.m. Asterisks denote values significantly different from the respective controls (P<0.05). N=7–8 except for the lowest (10–25 µmol l–1; N=3) and the highest (1000–2000 µmol l–1, N=4–5) Cd levels.

 

Figure 3
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  		Fig. 3. Effects of cadmium (Cd) exposure on glutathione synthesis rates in gill and hepatopancreas (HP) cells of oysters estimated as the rates of cycloheximide-insensitive incorporation of [3H]Gly. Mean values of glutathione synthesis in the cells exposed to different Cd levels (x-axis) for 4 h at 20°C are expressed as % of the respective controls. Vertical bars represent s.e.m. The asterisk denotes values significantly different from the respective control [P<0.01; the dagger denotes a marginally significant difference (P=0.09)]. N=5.

 

Figure 4
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  		Fig. 4. Effects of cadmium (Cd) exposure on total glutathione (GSH) concentrations in gill (A) and hepatopancreas (B) cells of oysters. Cells were incubated for varying periods of time (x-axis) with 0 µmol l–1 (control) or 2000 µmol l–1 Cd at 20°C, and their total (reduced + oxidized) GSH levels were determined. P-values for the effect of Cd on GSH levels, as determined by ANOVA, are given. At each exposure time, differences in GSH content between control and Cd-exposed cells were not significant (P>0.05) but the overall effect of Cd estimated by ANOVA was significant in hepatopancreas cells (see P-values in the figures). Vertical bars represent s.e.m. (N=5).

 

Figure 5
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  		Fig. 5. Effects of cadmium (Cd) exposure on expression of metallothionein mRNA in gill and hepatopancreas (HP) cells of oysters. Metallothionein levels were measured using real-time PCR, normalized to the actin mRNA levels and expressed as % of the respective controls. Cells were exposed to varying Cd levels (x-axis) for 4 h at 20°C. (A) mRNA for total metallothioneins and (B) for metallothionein I only. Vertical bars represent s.e.m. Asterisks denote values significantly different from the respective controls (P<0.05). N=9.

 

Figure 6
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  		Fig. 6. Effects of cadmium (Cd) exposure on expression of cytoplasmic (HSP70, HSP90) and mitochondrial (HSP60) heat shock proteins in gill and hepatopancreas (HP) cells of oysters. HSP levels were measured using immunoblotting and expressed as % of the respective controls. Cells were exposed to varying Cd levels (x-axis) for 4 h at 20°C. Vertical bars represent s.e.m. Asterisks denote values significantly different from the respective controls (P<0.05). N=5.

 




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