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Biology

Bir Zebra balığı Larva Lusıferaz Muhabir Sistemi ile glukokortikoid Signaling'den için bir Kimyasal Tarama Prosedürü

doi: 10.3791/50439 Published: September 10, 2013

Materials

Name Company Catalog Number Comments
Buffer composition + reagents
Dimethyl sulphoxide (DMSO) Carl Roth GmbH Co KG A994.2
FDA approved drug library Enzo Life Sciences BML-2841-0100
Luciferin Biosynth L-8220
Dexamethasone Sigma-Aldrich D4902
Methylene Blue Sigma-Aldrich M9140
E3 N/A N/A 5 mM NaCl, 0.17 mM KCl, 0.33 mM CaCl2
Instruments
Multiprobe II PerkinElmer 8 channel, equipped with disposable tip adaptor
Liquidator 96 Steinbrenner Laborsysteme hand-operated 96-channel pipette
EnVision XCite Multilabel Plate Reader PerkinElmer Equipped with stacker automation, temperature control, barcode reader and enhanced luminescence detector
Plasticware + consumables
96-Well Storage Plate ABgene AB-0765 round well, 0.8 ml
Cover films, ratiolab 6018412 self adhesive, DMSO resistent
Pipette tips Steinbrenner Laborsysteme LRF-200L for liquidator 96, 200 μl, low retention
TopSeal-A PerkinElmer 6005185
OptiPlate-96 PerkinElmer 6005299 white opaque 96-well microplate
Barcode Labels PerkinElmer 1608182
filtered polypropylene IsoTip pipette tips Corning S058.4809
Animals
GRE:Luc fish N/A ZDB-TGCONSTRCT-120920-1 available at the European Zebrafish Resource Centre EZRC, Karlsruhe

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References

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  2. Kassel, O., Herrlich, P. Crosstalk between the glucocorticoid receptor and other transcription factors: molecular aspects. Mol Cell Endocrinol. 275, 13-29 (2007).
  3. Baschant, U., Tuckermann, J. The role of the glucocorticoid receptor in inflammation and immunity. J Steroid Biochem Mol Biol. 120, 69-75 (2010).
  4. Schacke, H., Docke, W. D., Asadullah, K. Mechanisms involved in the side effects of glucocorticoids. Pharmacol Ther. 96, 23-43 (2002).
  5. De Bosscher, K. Selective Glucocorticoid Receptor modulators. J Steroid Biochem Mol Biol. 120, 96-104 (2010).
  6. Kaufman, C. K., White, R. M., Zon, L. Chemical genetic screening in the zebrafish embryo. Nat Protoc. 4, 1422-1432 (2009).
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  8. Casals-Casas, C., Desvergne, B. Endocrine disruptors: from endocrine to metabolic disruption. Annu Rev Physiol. 73, 135-162 (2011).
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  10. Odermatt, A., Gumy, C., Atanasov, A. G., Dzyakanchuk, A. A. Disruption of glucocorticoid action by environmental chemicals: potential mechanisms and relevance. J Steroid Biochem Mol Biol. 102, 222-231 (2006).
  11. Odermatt, A., Gumy, C. Glucocorticoid and mineralocorticoid action: why should we consider influences by environmental chemicals. Biochem Pharmacol. 76, 1184-1193 (2008).
  12. Lohr, H., Hammerschmidt, M. Zebrafish in endocrine systems: recent advances and implications for human disease. Annu Rev Physiol. 73, 183-211 (2011).
  13. Schaaf, M. J. Discovery of a functional glucocorticoid receptor beta-isoform in zebrafish. Endocrinology. 149, 1591-1599 (2008).
  14. Schaaf, M. J., Chatzopoulou, A., Spaink, H. P. The zebrafish as a model system for glucocorticoid receptor research. Comp Biochem Physiol A Mol Integr Physiol. 153, 75-82 (2009).
  15. Alsop, D., Vijayan, M. The zebrafish stress axis: molecular fallout from the teleost-specific genome duplication event. Gen Comp Endocrinol. 161, 62-66 (2009).
  16. Alsop, D., Vijayan, M. M. Molecular programming of the corticosteroid stress axis during zebrafish development. Comp Biochem Physiol A Mol Integr Physiol. 153, 49-54 (2009).
  17. Schoonheim, P. J., Chatzopoulou, A., Schaaf, M. J. The zebrafish as an in vivo model system for glucocorticoid resistance. Steroids. 75, 918-925 (2010).
  18. Weger, B. D., Weger, M., Nusser, M., Brenner-Weiss, G., Dickmeis, T. A Chemical Screening System for Glucocorticoid Stress Hormone Signaling in an Intact Vertebrate. ACS Chem Biol. 7, (2012).
  19. Hirota, T. A chemical biology approach reveals period shortening of the mammalian circadian clock by specific inhibition of GSK-3beta. Proc Natl Acad Sci U S A. 105, 20746-20751 (2008).
  20. Birmingham, A. Statistical methods for analysis of high-throughput RNA interference screens. Nat Methods. 6, 569-575 (2009).
  21. Warnes, G. R., Bolker, B., Bonebakker, L., Gentleman, R., Liaw, W. H. A. gplots: Various R programming tools for plotting data. 2.11.0, (2012).
  22. Carey, V., Redestig, H. ROC: utilities for ROC, with uarray focus. Boston, MA, USA. (2013).
  23. Fan, F., Wood, K. V. Bioluminescent assays for high-throughput screening. Assay Drug Dev Technol. 5, 127-136 (2007).
  24. Graf, S. F., Hotzel, S., Liebel, U., Stemmer, A., Knapp, H. F. Image-based fluidic sorting system for automated Zebrafish egg sorting into multiwell plates. J Lab Autom. 16, 105-111 (2011).
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Bir Zebra balığı Larva Lusıferaz Muhabir Sistemi ile glukokortikoid Signaling'den için bir Kimyasal Tarama Prosedürü
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Cite this Article

Weger, B. D., Weger, M., Jung, N., Lederer, C., Bräse, S., Dickmeis, T. A Chemical Screening Procedure for Glucocorticoid Signaling with a Zebrafish Larva Luciferase Reporter System. J. Vis. Exp. (79), e50439, doi:10.3791/50439 (2013).More

Weger, B. D., Weger, M., Jung, N., Lederer, C., Bräse, S., Dickmeis, T. A Chemical Screening Procedure for Glucocorticoid Signaling with a Zebrafish Larva Luciferase Reporter System. J. Vis. Exp. (79), e50439, doi:10.3791/50439 (2013).

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