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Bioenergetic Analyses of In Vitro and In Vivo Samples to Guide Toxicological Endpoints

  • Jonathan W. BoydEmail author
  • Julia A. Penatzer
  • Nicole Prince
  • Julie V. Miller
  • Alice A. Han
  • Holly N. Currie
Protocol
  • 181 Downloads
Part of the Methods in Molecular Biology book series (MIMB, volume 2102)

Abstract

Toxicology is a broad field that requires the translation of biochemical responses to xenobiotic exposures into useable information to ensure the safety of the public. Modern techniques are improving rapidly, both quantitatively and qualitatively, to provide the tools necessary to expand available toxicological datasets and refine our ability to translate that data into relevant information via bioinformatics. These new techniques can, and do, impact many of the current critical roles in toxicology, including the environmental, forensic, preclinical/clinical, and regulatory realms. One area of rapid expansion is our understanding of bioenergetics, or the study of the transformation of energy in living organisms, and new mathematical approaches are needed to interpret these large datasets. As bioenergetics are intimately involved in the regulation of how and when a cell responds to xenobiotics, monitoring these changes (i.e., metabolic fluctuations) in cells/tissues post-exposure provides an approach to define the temporal scale of pharmacodynamic responses, which can be used to guide additional toxicological techniques (e.g., “omics”). This chapter will summarize important in vitro assays and in vivo imaging techniques to take real-time measurements. Using this information, our laboratory has utilized bioenergetics to identify significant time points of pharmacodynamic relevance as well as forecast the cell’s eventual fate.

Key words

Bioenergetics ATP MitoXpress Live/Dead NADH 2-NBDG ICG 18FDG Forecasting 

References

  1. 1.
    Bioenergetics (n.d.). In Merriam-Webster online. https://www.merriam-webster.com/dictionary/bioenergetics. Accessed 10 Dec 2018
  2. 2.
    Miller J, Prince N, Mouch J, Boyd J (2018) A toxicological application of signal transduction: early cellular changes can be indicative of toxicity. In: Boyd J, Neubig R (eds) In cellular signal transduction in toxicology and pharmacology: data collection, analysis, and interpretation. John Wiley & Sons, Ltd, ChichesterGoogle Scholar
  3. 3.
    Hynes J, Carey C, Will Y (2016) Fluorescence-based microplate assays for in vitro assessment of mitochondrial toxicity, metabolic perturbation, and cellular oxygenation. Curr Protoc Toxicol 70:1–30CrossRefGoogle Scholar
  4. 4.
    Anusree SS, Nisha VM, Priyanka A, Raghu KG (2015) Insulin resistance by TNF-α is associated with mitochondrial dysfunction in 3T3-L1 adipocytes and is ameliorated by punicic acid, a PPARγ agonist. Mol Cell Endocrinol 413:120–128CrossRefGoogle Scholar
  5. 5.
    Morten KJ, Badder L, Knowles HJ (2013) Differential regulation of HIF-mediated pathways increases mitochondrial metabolism and ATP production in hypoxic osteoclasts. J Pathol 229:755–764CrossRefGoogle Scholar
  6. 6.
    MitoXpress Xtra Oxygen Consumption Assay (2018). https://www.agilent.com/cs/library/usermanuals/public/MitoXpress_Xtra_Oxygen_Consumption_Assay.pdf. Accessed 10 Dec 2018
  7. 7.
    Calcein AM (2018). https://www.thermofisher.com/order/catalog/product/C3099. Accessed 11 Dec 2018
  8. 8.
    Cell Staining (2018). https://www.dojindo.com/Protocol/CellStaining_Protocol.pdf. Accessed 17 Dec 2018
  9. 9.
  10. 10.
    Bratosin D, Mitrofan L, Palii C, Estaquier J, Montreuil J (2005) Novel fluorescence assay using calcein-AM for the determination of human erythrocyte viability and aging. Cytometry A 66A:78–84CrossRefGoogle Scholar
  11. 11.
    Ohno Y, Fujita K, Toyofuku T, Nakamaura T (2016) Cytological observations of the large symbiotic foraminifer amphisorus kudakajimensis using calcein acetoxymethyl ester. PLoS One 11:e0165844CrossRefGoogle Scholar
  12. 12.
    Live/Dead Viability/Cytotoxicity Kit (2005). https://assets.thermofisher.com/TFS-Assets/LSG/manuals/mp03224.pdf. Accessed 10 Dec 2018
  13. 13.
    Martinez-Madrid B, Dolmans MM, Van Langendonckt A, Defrere S, Van Eyck AS, Donnez J (2004) Ficoll density gradient method for recovery of isolated human ovarian primordial follicles. Fertil Steril 82:1648–1653CrossRefGoogle Scholar
  14. 14.
    Sanfilippo S, Canis M, Ouchchane L, Botchorishvili R, Artonne C, Janny L, Brugnon F (2011) Viability assessment of fresh and frozen/thawed isolated human follicles: reliability of two methods (Trypan blue and Calcein AM/ethidium homodimer-1). J Assist Reprod Genet 28:1151–1156CrossRefGoogle Scholar
  15. 15.
    Glazer AN, Peck K, Mathies RA (1990) A stable double-stranded DNA-ethidium homodimer complex: application to picogram fluorescence detection of DNA in agarose gels. Proc Natl Acad Sci U S A 87:3851–3855CrossRefGoogle Scholar
  16. 16.
    Stocks SM (2004) Mechanism and use of the commercially available viability stain, BacLight. Cytometry A 61A:189–195CrossRefGoogle Scholar
  17. 17.
    Osellame LD, Blacker TS, Duchen MR (2012) Cellular and molecular mechanisms of mitochondrial function. Best Pract Res Clin Endocrinol Metab 26:711–723CrossRefGoogle Scholar
  18. 18.
    Yang Y, Sauve AA (2016) NAD+ metabolism: bioenergetics, signaling and manipulation for therapy. Biochim Biophys Acta 1864:1787–1800CrossRefGoogle Scholar
  19. 19.
    TeSlaa T, Teitell MA (2014) Techniques to monitor glycolysis. Methods Enzymol 542:91–114CrossRefGoogle Scholar
  20. 20.
    Zou C, Wang Y, Shen Z (2005) 2-NBDG as a fluorescent indicator for direct glucose uptake measurement. J Biochem Biophys Methods 64:207–215CrossRefGoogle Scholar
  21. 21.
    O’Neil RG, Wu L, Mullani N (2005) Uptake of a fluorescent deoxyglucose analog (2-NBDG) in tumor cells. Mol Imaging Biol 7:388–392CrossRefGoogle Scholar
  22. 22.
    Taylor K, Lemon JA, Boreham DR (2014) Radiation-induced DNA damage and the relative biological effectiveness of 18F-FDG in wild-type mice. Mutagenesis 29:279–287CrossRefGoogle Scholar
  23. 23.
    Guo Y, Gao F, Wang S, Ding Y, Zhang H, Wang J, Ding MP (2009) In vivo mapping of temporospatial changes in glucose utilization in rat brain during epileptogenesis: an 18F-fluorodeoxyglucose–small animal positron emission tomography study. Neuroscience 162:972–979CrossRefGoogle Scholar
  24. 24.
    Mateo J, Bilbao I, Vaquero JJ, Ruiz-Cabello J, Espana S (2015) In vivo 18F-FDG-PET imaging in mouse atherosclerosis. Methods Mol Biol 1339:377–386CrossRefGoogle Scholar
  25. 25.
    Boni L, David G, Mangano A, Dionigi G, Rausei S, Spampatti S et al (2015) Clinical applications of indocyanine green (ICG) enhanced fluorescence in laparoscopic surgery. Surg Endosc 29:2046–2055CrossRefGoogle Scholar
  26. 26.
    Okumura K, Yoshida K, Yoshioka K, Aki S, Yoneda N, Inoue D et al (2018) Photoacoustic imaging of tumour vascular permeability with indocyanine green in a mouse model. Eur Radiol Exp 2:5CrossRefGoogle Scholar
  27. 27.
    Jiang JX, Keating JJ, De Jesus EM, Judy RP, Madajewski B, Venegas O, Okusanya OT, Singhal S (2015) Optimization of the enhanced permeability and retention effect for near-infrared imaging of solid tumors with indocyanine green. Am J Nucl Med Mol Imaging 5:390–400PubMedPubMedCentralGoogle Scholar
  28. 28.
    Portnoy E, Vakruk N, Bishara A, Shmuel M, Magdassi S, Golenser J, Eyal S (2016) Indocyanine green liposomes for diagnosis and therapeutic monitoring of cerebral malaria. Theranostics 6:167–176CrossRefGoogle Scholar
  29. 29.
    Dietz MJ, Hare JT, Ueno C, Prud’homme BJ, Boyd JW (2018) Laser-assisted fluorescent angiography to assess tissue perfusion in the setting of traumatic elbow dislocation. Wounds 30:E93–E97PubMedGoogle Scholar
  30. 30.
    Yoneya S, Saito T, Komatsu Y, Koyama I, Takahashi K, Duvoll-Young J (1998) Binding properties of indocyanine green in human blood. IOVS 39:1286–1290Google Scholar
  31. 31.
    Alander JT, Kaartinen I, Laakso A, Patila T, Spillmann T, Tuchin VV, Venermo M, Valisuo P (2012) A review of indocyanine green fluorescent imaging in surgery. Int J Biomed Imaging 2012:940585CrossRefGoogle Scholar
  32. 32.
    Vrana JA, Currie HN, Han AA, Boyd J (2014) Forecasting cell death dose-response from early signal transduction responses in vitro. Toxicol Sci 140:338–351CrossRefGoogle Scholar
  33. 33.
    Kadenbach B (1986) Regulation of respiration and ATP synthesis in higher organisms: hypothesis. J Bioenerg Biomembr 18:39–54CrossRefGoogle Scholar
  34. 34.
    Nelson DL, Cox MM (2008) Lehninger principles of biochemistry. W. H. Freeman, New YorkGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Jonathan W. Boyd
    • 1
    • 2
    • 3
    Email author
  • Julia A. Penatzer
    • 1
  • Nicole Prince
    • 1
  • Julie V. Miller
    • 4
  • Alice A. Han
    • 5
  • Holly N. Currie
    • 6
  1. 1.Department of OrthopaedicsWest Virginia University School of MedicineMorgantownUSA
  2. 2.Department of Physiology and PharmacologyWest Virginia University School of MedicineMorgantownUSA
  3. 3.Department of Occupational and Environmental Health SciencesWest Virginia University School of Public HealthMorgantownUSA
  4. 4.Cardno ChemRiskPittsburghUSA
  5. 5.Department of Chemistry, West Virginia UniversityMorgantownUSA
  6. 6.Frostburg State UniversityFrostburgUSA

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