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Double-labelling methods used to diagnose apoptotic and necrotic cell degradations in copepod nauplii

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Abstract

Two double-labelling methods, Tunel+propidium iodide and Annexin V-FITC+propidium iodide, have been tested to diagnose cell degradation processes in N1–N2 nauplius stages produced by spawning females of the copepod Calanus helgolandicus fed either non-toxic (PRO: control) or toxic (TR1) diets under laboratory conditions. Observation of labelled samples with a confocal laser-scanning microscope revealed that the maternal-food effect, following absorption of the toxic diet, induced cell apoptosis and necrosis in 80–100% of the offspring, which sooner or later died. Similar cell damages were also observed in 18–50% of the nauplii produced by females incubated in filtered seawater. Such cell degradation processes and high mortality were not observed in nauplii produced by females fed the non-toxic diet. Our protocols allowed detection of cell degradations before death of unhealthy larvae, preferentially with a confocal microscope or a conventional visible-fluorescent-light microscope.

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Acknowledgements

We are grateful to the sailors at the Roscoff Marine Station for collecting plankton at sea. Comments from anonymous referees, greatly help to improve this paper. This work was supported by CNRS and by the French PNEC-ART 2 programme.

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Correspondence to S. A. Poulet.

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Communicated by O. Kinne, Oldendorf/Luhe

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Poulet, S.A., Richer de Forges, M., Cueff, A. et al. Double-labelling methods used to diagnose apoptotic and necrotic cell degradations in copepod nauplii. Marine Biology 143, 889–895 (2003). https://doi.org/10.1007/s00227-003-1142-4

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