Whole-Organism Screening for Modulators of Fasting Metabolism Using Transgenic Zebrafish

Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1263)

Abstract

Organismal energy homeostasis is maintained by complex interorgan communications making the discovery of novel drugs against metabolic diseases challenging using traditional high-throughput approaches in vitro. Here, we describe a method that rapidly identifies small molecules with an impact on organismal energy balance in vivo. Specifically, we developed a whole-organism screen for modulators of fasting metabolism using transgenic bioluminescence-reporter zebrafish for the gluconeogenic gene phosphoenolpyruvate-carboxykinase 1 (pck1).

Key words

Zebrafish Gluconeogenesis Diabetes Metabolic disease Fasting metabolism Small-molecule screening Bioluminescence Phosphoenolpyruvate-carboxykinase pck1 Transgenesis 

Notes

Acknowledgements

This study was supported in part by a postdoctoral fellowship DFG GU 1082/101 from the German Research Foundation to P.G., NIH grant RO1 DK60322, a pilot and feasibility award from the University of California—San Francisco diabetes center funded by NIH U01 DK089541 and the Packard Foundation to D.Y.R.S.

References

  1. 1.
    Baur JA et al (2006) Resveratrol improves health and survival of mice on a high-calorie diet. Nature 444:337–342PubMedCrossRefGoogle Scholar
  2. 2.
    Le Couteur DG et al (2012) Aging biology and novel targets for drug discovery. J Gerontol A Biol Sci Med Sci 67:168–174PubMedCrossRefGoogle Scholar
  3. 3.
    Guarente LL (2006) Sirtuins as potential targets for metabolic syndrome. Nature 444:868–874PubMedCrossRefGoogle Scholar
  4. 4.
    Anson RM et al (2003) Intermittent fasting dissociates beneficial effects of dietary restriction on glucose metabolism and neuronal resistance to injury from calorie intake. Proc Natl Acad Sci U S A 100:6216–6220PubMedCentralPubMedCrossRefGoogle Scholar
  5. 5.
    Moller DE (2012) Metabolic disease drug discovery- “hitting the target” is easier said than done. Cell Metab 15:19–24PubMedCrossRefGoogle Scholar
  6. 6.
    Lemieux GA et al (2011) A whole-organism screen identifies new regulators of fat storage. Nat Chem Biol 7:206–213PubMedCentralPubMedCrossRefGoogle Scholar
  7. 7.
    Gut P et al (2013) Whole-organism screening for gluconeogenesis identifies activators of fasting metabolism. Nat Chem Biol 9:97–104PubMedCentralPubMedCrossRefGoogle Scholar
  8. 8.
    Andersson O et al (2012) Adenosine signaling promotes regeneration of pancreatic β cells in vivo. Cell Metab 15:885–894PubMedCentralPubMedCrossRefGoogle Scholar
  9. 9.
    Zon LI, Peterson RT (2005) In vivo drug discovery in the zebrafish. Nat Rev Drug Discov 4:35–44PubMedCrossRefGoogle Scholar
  10. 10.
    Lin HV, Accili D (2011) Hormonal regulation of hepatic glucose production in health and disease. Cell Metab 14:9–19PubMedCentralPubMedCrossRefGoogle Scholar
  11. 11.
    Rena G, Pearson ER, Sakamoto K (2013) Molecular mechanism of action of metformin: old or new insights? Diabetologia 56:1898–1906PubMedCentralPubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  1. 1.Gladstone Institutes and University of CaliforniaSan FranciscoUSA
  2. 2.Nestlé Institute of Health Sciences S.A.LausanneSwitzerland
  3. 3.Max-Planck Institute for Heart and Lung ResearchBad NauheimGermany

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