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Metabolic Programming of Zebrafish, Danio rerio
Physiological performance as explained by Dynamic Energy Budget Theory and life-cycle consequences of uranium induced perturbations
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Starrlight Augustine
1Promoteren: Prof.Dr.Sebastiaan A.L.M. Kooijman, Prof.Dr. Christelle Adam-GuillerminCopromotor: Dr. Béatrice Gagnaire
Extinction
Context : Environmental Risk Assessment
Aquatic ecosystems are the recipient of byproducts of our industries:
How much of each type of compound can be released into the environment without inducing deleterious side effects?
U
UU
U
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Extinction
Context : Environmental Risk Assessment
Aquatic ecosystems are the recipient of byproducts of our industries:
How much of each type of compound can be released into the environment without inducing deleterious side effects?
U
UU
U
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Context: IRSN research programme ENVIRHOM-ECO
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
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Mission statement: Evaluate long term effects of chronic exposure to radionuclides on the ecosystem.
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From molecules to ecosystems …
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SpaceSpace
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From molecules to ecosystems …
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SpaceSpace
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From molecules to ecosystems …
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SpaceSpace
Growth + reproduction
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OECD test organism: zebrafish
>90 jours2-3 cm
5 jours0.45 cm
Embryo
Juvenile
Adult
BIRTHPUBERTY
>90 days2-3 cm
5 days0.45 cm
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Dynamic Energy Budget (DEB) Theory
Internal concen-tration
Enough Too much
NEC
Effects
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.
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NEC = No effect Internal Concentration
0
0
Too little
Phase I: characterizing the physiological performance of non-exposed zebrafish (the blank)
Growth and Reproduction at different food levels
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Literature data
Zebrafish DEB model
Ageing
Temperature
Starvation8
Results: metabolic programming uncovered
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Processes specified by the DEB model
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Augustine et al 2011aAugustine et al 2011bMueller et al 2012
Results: metabolic programming uncovered
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Feeding
Processes specified by the DEB model
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Augustine et al 2011aAugustine et al 2011bMueller et al 2012
Results: metabolic programming uncovered
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Feeding
Digestion
Processes specified by the DEB model
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Augustine et al 2011aAugustine et al 2011bMueller et al 2012
Results: metabolic programming uncovered
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Feeding
Digestion
Storage
Processes specified by the DEB model
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Augustine et al 2011aAugustine et al 2011bMueller et al 2012
Results: metabolic programming uncovered
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Feeding
Digestion
Storage
MaintenanceProcesses specified by the DEB model
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Augustine et al 2011aAugustine et al 2011bMueller et al 2012
Results: metabolic programming uncovered
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Feeding
Digestion
Storage
Maintenance
Growth
Processes specified by the DEB model
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Augustine et al 2011aAugustine et al 2011bMueller et al 2012
Results: metabolic programming uncovered
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Feeding
Digestion
Storage
Maintenance
Growth
Development
Processes specified by the DEB model
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Augustine et al 2011aAugustine et al 2011bMueller et al 2012
Results: metabolic programming uncovered
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Feeding
Digestion
Storage
Maintenance
Growth
Development
Processes specified by the DEB model
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Augustine et al 2011aAugustine et al 2011bMueller et al 2012
Reproduction
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Feeding
Digestion
Storage
Growth
Development
Processes specified by the DEB model
Reproduction
Ageing
Maintenance
Results: metabolic programming uncovered
Augustine et al 2011aAugustine et al 2011bMueller et al 2012
Phase II: characterizing uranium induced deviations from the physiological performance of the blank
+
Literature data
Zebrafish DEB model
0 nM U 84 nM U 420 nM U
Growth and Reproduction at three levels of uranium
Uptake and elimination
How effects depend on internal concentration
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Results: effects of uranium
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Uranium induces toxic effects almost as soon as it is internalized
Effects on -growth (early life stages) -maintenance (adults)
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Feeding
Digestion
Storage
Growth
Development
Processes specified by the DEB model
Reproduction
Ageing
Maintenance
Results: effects of uranium
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Dry mass (µg)
Age, d
Data: Stéphanie Bourrachot 2009Lines: DEB model predictions
1054 nM
0 nM
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Feeding
Digestion
Storage
Growth
Development
Processes specified by the DEB model
Reproduction
Ageing
Maintenance
Results: effects of uranium
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Allocation to reproduction is a continuous process
Feeding
Digestion
Storage
Growth
Development
Processes specified by the DEB model
Reproduction
Ageing
Maintenance
Results: effects of uranium
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Allocation to reproduction is a continuous process
Spawning is discontinuous, but when the organism does not spawn the mass invested in reproduction remains inside the individual
Feeding
Digestion
Storage
Growth
Development
Processes specified by the DEB model
Reproduction
Ageing
Maintenance
Results: effects of uranium
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Allocation to reproduction is a continuous process
Spawning is discontinuous, but when the organism does not spawn the mass invested in reproduction remains inside the individual
When energy assimilated from food no longer covers maintenance then missing energy is taken from that invested in reproduction
Feeding
Digestion
Storage
Growth
Development
Processes specified by the DEB model
Reproduction
Ageing
Maintenance
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140 10 20 30 400
500
1000
Experiment time, days
Cumulated number of eggs
Augustine et al 2012
Spawning
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140 10 20 30 400
500
1000
Experiment time, days
866 eggs spawned in total is about 0.36 g (fresh weight)
Cumulated number of eggs
Augustine et al 2012
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0.4
0.6
0.8
1
1.2
0 10 20 30 400
500
1000
Experiment time, days
Wet mass, g
Final observed mass
DEB model prediction for the blank
Cumulated number of eggs
Augustine et al 2012
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0.4
0.6
0.8
1
1.2About half the final mass of the fish was spawned !!
0 10 20 30 400
500
1000
Experiment time, days
Wet mass, g
Cumulated number of eggs
Final observed mass
DEB model prediction for the blank
Augustine et al 2012
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0.4
0.6
0.8
1
1.2
0 10 20 30 400
500
1000
Experiment time, days
Cumulated number of eggs
Wet mass, g
Final observed mass
DEB model prediction for the blank
DEB model prediction for increasing maintenance costs
Augustine et al 2012
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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
From individual to ecosystems …
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SpaceSpace
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From individual to molecules …
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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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