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Metabolic Programming of Zebrafish, Danio rerio

Metabolic Programming of Zebrafish, Danio rerio. Physiological performance as explained by Dynamic Energy Budget Theory and life-cycle consequences of uranium induced perturbations. Starrlight Augustine. Promoteren: Prof.Dr.Sebastiaan A.L.M. Kooijman, Prof.Dr. Christelle Adam-Guillermin

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Metabolic Programming of Zebrafish, Danio rerio

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  1. MetabolicProgramming of Zebrafish, Danio rerio Physiological performance as explained by Dynamic Energy Budget Theory and life-cycle consequences of uranium induced perturbations Starrlight Augustine Promoteren: Prof.Dr.Sebastiaan A.L.M. Kooijman, Prof.Dr. Christelle Adam-Guillermin Copromotor: Dr. Béatrice Gagnaire

  2. Context : Environmental Risk Assessment Aquatic ecosystems are the recipient of byproducts of our industries: April 23, 2012 - Public Doctoral Defense – Vrije Universiteit U Extinction How much of each type of compound can be released into the environment without inducing deleterious side effects? U U U

  3. Context : Environmental Risk Assessment Aquatic ecosystems are the recipient of byproducts of our industries: April 23, 2012 - Public Doctoral Defense – Vrije Universiteit U Extinction How much of each type of compound can be released into the environment without inducing deleterious side effects? U U U

  4. Context: IRSN research programme ENVIRHOM-ECO April 23, 2012 - Public Doctoral Defense – Vrije Universiteit Mission statement: Evaluate long term effects of chronic exposure to radionuclides on the ecosystem. Major cities Mine tailing storage sites still in use in 1989 Uranium mines and mine tailings storage in France Mine tailing storage closed in in 1989

  5. Frommolecules to ecosystems … Space

  6. Frommolecules to ecosystems … Space

  7. Frommolecules to ecosystems … Growth + reproduction Space

  8. OECD test organism: zebrafish April 23, 2012 - Public Doctoral Defense – Vrije Universiteit Adult Embryo Juvenile PUBERTY BIRTH >90 days 2-3 cm 5 days 0.45 cm >90 jours 2-3 cm 5 jours 0.45 cm

  9. Effects Enough Too much Internal concen-tration NEC DynamicEnergy Budget (DEB) Theory General unified theory in biology created over 30 years ago (Kooijman 1993, 2000, 2010) which specifies the acquisition and use of energy for all organisms. The theory respects mass and energy balancing. April 23, 2012 - Public Doctoral Defense – Vrije Universiteit Too little 0 0 NEC = No effect Internal Concentration

  10. Phase I: characterizing the physiological performance of non-exposed zebrafish (the blank) Growth and Reproduction at different food levels Literature data + Temperature Zebrafish DEB model Starvation Ageing

  11. Results: metabolicprogramminguncovered Processes specified by the DEB model Augustine et al 2011a Augustine et al 2011b Mueller et al 2012

  12. Results: metabolicprogramminguncovered Feeding Processes specified by the DEB model Augustine et al 2011a Augustine et al 2011b Mueller et al 2012

  13. Results: metabolicprogramminguncovered Feeding Digestion Processes specified by the DEB model Augustine et al 2011a Augustine et al 2011b Mueller et al 2012

  14. Results: metabolicprogramminguncovered Feeding Digestion Storage Processes specified by the DEB model Augustine et al 2011a Augustine et al 2011b Mueller et al 2012

  15. Results: metabolicprogramminguncovered Feeding Digestion Storage Maintenance Processes specified by the DEB model Augustine et al 2011a Augustine et al 2011b Mueller et al 2012

  16. Results: metabolicprogramminguncovered Feeding Digestion Storage Maintenance Processes specified by the DEB model Growth Augustine et al 2011a Augustine et al 2011b Mueller et al 2012

  17. Results: metabolicprogramminguncovered Feeding Digestion Storage Maintenance Processes specified by the DEB model Growth Development Augustine et al 2011a Augustine et al 2011b Mueller et al 2012

  18. Results: metabolicprogramminguncovered Feeding Digestion Storage Maintenance Processes specified by the DEB model Growth Development Reproduction Augustine et al 2011a Augustine et al 2011b Mueller et al 2012

  19. Results: metabolicprogramminguncovered Feeding Digestion Storage Maintenance Processes specified by the DEB model Growth Development Reproduction Augustine et al 2011a Augustine et al 2011b Mueller et al 2012 Ageing

  20. Phase II: characterizing uranium induced deviations from the physiological performance of the blank Growth and Reproduction at three levels of uranium Literature data + 0 nM U 84 nM U 420 nM U Zebrafish DEB model Uptake and elimination How effects depend on internal concentration

  21. Results: effects of uranium Feeding Uranium induces toxic effects almost as soon as it is internalized Digestion Storage • Effects on • -growth (early life stages) • maintenance (adults) Maintenance Processes specified by the DEB model Growth Development Reproduction Ageing

  22. Results: effects of uranium Feeding 0 nM Dry mass (µg) Digestion 1054 nM Storage Maintenance Processes specified by the DEB model Growth Development Age, d Reproduction Data: StéphanieBourrachot 2009 Lines: DEB model predictions Ageing

  23. Results: effects of uranium Feeding Digestion Allocation to reproduction is a continuous process Storage Maintenance Processes specified by the DEB model Growth Development Reproduction Ageing 2000 1.2 1500 1 0.8 1000 0.6 500 0.4 0 0.2 0 10 20 30 40

  24. Results: effects of uranium Feeding Digestion Allocation to reproduction is a continuous process Storage Spawning is discontinuous, but when the organism does not spawn the mass invested in reproduction remains inside the individual Maintenance Processes specified by the DEB model Growth Development Reproduction Ageing 2000 1.2 1500 1 0.8 1000 0.6 500 0.4 0 0.2 0 10 20 30 40

  25. Results: effects of uranium Feeding Digestion Allocation to reproduction is a continuous process Storage Spawning is discontinuous, but when the organism does not spawn the mass invested in reproduction remains inside the individual Maintenance Processes specified by the DEB model Growth Development When energy assimilated from food no longer covers maintenance then missing energy is taken from that invested in reproduction Reproduction Ageing 2000 1.2 1500 1 0.8 1000 0.6 500 0.4 0 0.2 0 10 20 30 40

  26. Spawning Cumulated number of eggs 1000 500 0 0 10 20 30 40 Experiment time, days Augustine et al 2012

  27. 866 eggs spawned in total is about 0.36 g (fresh weight) Cumulated number of eggs 1000 500 0 0 10 20 30 40 Experiment time, days Augustine et al 2012

  28. Wet mass, g 1.2 Cumulated number of eggs Final observed mass 1 DEB model prediction for the blank 0.8 1000 0.6 500 0.4 0 0 10 20 30 40 Experiment time, days Augustine et al 2012

  29. Wet mass, g 1.2 Cumulated number of eggs Final observed mass 1 About half the final mass of the fish was spawned !! DEB model prediction for the blank 0.8 1000 0.6 500 0.4 0 0 10 20 30 40 Experiment time, days Augustine et al 2012

  30. Wet mass, g Cumulated number of eggs 1.2 Final observed mass 1 DEB model prediction for the blank DEB model prediction for increasing maintenance costs 0.8 1000 0.6 500 0.4 0 0 10 20 30 40 Experiment time, days Augustine et al 2012

  31. How do these results link back to the initial problem ? • Uranium turns out to be toxic and effects show up as soon as it is internalized. • The zebrafish DEB model is a very sensitive tool for detecting effects • The model is also useful for interpreting data and comparing studies • Effects on populations and ecosystems follow from effects on individuals

  32. Fromindividual to ecosystems … Fromindividual to molecules … Space

  33. Thank you for your attention Thanks are due to: Magalie Floriani (TEM) Virginie Cavalie (ICP-AES, TEM) Sylive Pierrisnard (ILC) Sandrine Pereira (immuno-histochemistry) Alban Carlat (data collection) « LRE » Laboratory and many more …

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