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First published online January 5, 2005
Journal of Experimental Biology 208, 277-285 (2005)
Published by The Company of Biologists 2005
doi: 10.1242/jeb.01368

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Symbiosis-induced adaptation to oxidative stress

Sophie Richier¹, Paola Furla¹, Amandine Plantivaux¹, Pierre-Laurent Merle¹ and Denis Allemand^1,2,*

¹ Université de Nice Sophia-Antipolis, BP 71, F-06108 Nice Cedex 02, France
² Centre Scientifique de Monaco, Avenue Saint-Martin, MC-98000 Monaco, Principality of Monaco

View larger version (55K): [in a new window]   Fig. 1. SOD electrophoretic patterns in symbiotic (Anemonia viridis) and non-symbiotic species (Actinia schmidti). (A) SOD activity in A. viridis animal (ectoderm plus endoderm) extract and (B) A. schmidti total extract in the absence (control) and in the presence of KCN (10 mmol l–1) revealed on native PAGE (8%) and by NBT staining. Arrows indicate pharmacological identification of bands: MnSOD (black arrows), CuZnSOD (open arrows). Cytosolic fractions containing a quantity of 150 µg of protein was loaded in each well. Similar results were obtained in four to 10 independent experiments for A. schmidti and A. viridis specimens, respectively.

View larger version (67K): [in a new window]   Fig. 2. Response of SOD activity to experimental hyperoxia. (A) SOD activity of the three separated compartments of A. viridis (Ec, ectoderm; Ed, endoderm; FIZ, freshly isolated zooxanthellae). (B) Total extract of A. schmidti in two conditions: control (C; air saturated) and hyperoxia (H,; O2-saturated seawater), revealed by native PAGE (8%) and NBT staining, either without KCN (–KCN) or with (+). Arrows indicate pharmacological identification of bands: MnSOD (black arrows), CuZnSOD (open arrows) and FeSOD (grey arrows). (d) Corresponds to an O2-induced SOD isoform (AsSODd). Cytosolic fractions containing a quantity of 150 µg of protein was loaded in each well. These patterns were reproducible in four independent experiments for each species.

View larger version (19K): [in a new window]   Fig. 3. Cellular damage in response to experimental hyperoxia. (A) Carbonyl and (B) MDA contents were measured in the three compartments of A. viridis (Ec, ectoderm; Ed, endoderm; FIZ, freshly isolated zooxanthellae) during the daytime. (C) Carbonyls and (D) MDA content were also measured in A. schmidti total extract during the daytime, for control (C) and hyperoxia-treated specimens (H). Cytosolic fractions containing a quantity of 100 and 150 µg of protein were analyzed for carbonyl and MDA content, respectively. Data are presented as means ± S.E.M. of four independent analyses. *Indicates significant differences between control and stressed specimens (Student's t-test P<0.05).

View larger version (69K): [in a new window]   Fig. 4. Response of SOD activity to +7°C increase above ambient. SOD activities of the three compartment of A. viridis (A) ectoderm, (B) endoderm and (C) FIZ (freshly isolated zooxanthellae), (D,E) total extract of A. schmidti under control conditions 0 (17°C) and after 1, 2 and 5 days of incubation to elevated temperature (24°C), revealed by native PAGE (8%) and NBT staining, either without KCN (–KCN) or with (+KCN). Pharmacological identification of the bands is indicated by arrows: MnSOD (black arrows), CuZnSOD (open arrows) and FeSOD (grey arrows). 150 µg of protein was loaded in each well. These patterns were reproducible in four independent experiments for each species.

View larger version (21K): [in a new window]   Fig. 5. Cellular damage in response to +7°C increase above ambient. (A) Carbonyls and (B) MDA content have been measured in the three compartments (Ec, ectoderm; Ed, endoderm; FIZ, freshly isolated zooxanthellae) of A. viridis and (C,D) in A. schmidti total extract. Control condition (white bars) and after 1 (grey bars), 2 (dark-grey bars) and 5 (black bars) days in +7°C seawater. Data are presented as means ± S.E.M. of four independent analyses. *Indicates significant differences between control and stress specimens (ANOVA, *P<0.05; P<0.01).

View larger version (57K): [in a new window]   Fig. 6. SOD activity in associated and dissociated symbiotic partners. SOD activity in animal compartments and FIZ of A. viridis (A) in control (C) and bleached conditions (Bl) and in total extract and FIZ of A. pulchella (B) for control (C), bleached specimen (Bl) and (FIZ) extracts, is revealed by 8% native PAGE and NBT staining. 150 µg of protein was loaded in each well. Arrows indicate pharmacological identification of bands: MnSOD (black arrows), CuZnSOD (open arrows) and FeSOD (grey arrows). These patterns were reproducible in four independent experiments for each species.

View larger version (79K): [in a new window]   Fig. 7. Interactions of symbiotic partners in SOD expression. SOD isoforms of the whole symbiotic association (T), the cultured zooxanthellae (CZ) and the freshly isolated zooxanthellae (FIZ) of two symbiotic organism A. viridis (Mediterrannean sea anemone) and Stylophora pistillata (tropical scleractinian coral) have been revealed by 8% native PAGE and NBT staining. Each lane was loaded with 150 µg of protein. Arrows indicate pharmacological identification of bands: MnSOD (black arrows), CuZnSOD (open arrows) and FeSOD (grey arrows). These patterns were reproducible in four independent experiments on each species.