Analytical and Bioanalytical Chemistry

, Volume 410, Issue 30, pp 7979–7986 | Cite as

IR-MALDESI mass spectrometry imaging of underivatized neurotransmitters in brain tissue of rats exposed to tetrabromobisphenol A

  • M. Caleb Bagley
  • Måns Ekelöf
  • Kylie Rock
  • Heather Patisaul
  • David C. MuddimanEmail author
Research Paper


There is a pressing need to develop tools for assessing possible neurotoxicity, particularly for chemicals where the mode of action is poorly understood. Tetrabromobisphenol A (TBBPA), a highly abundant brominated flame retardant, has lately been targeted for neurotoxicity analysis by concerned public health entities in the EU and USA because it is a suspected thyroid disruptor and neurotoxicant. In this study, infrared matrix-assisted laser desorption electrospray ionization (IR-MALDESI) coupled to a Q Exactive Plus mass spectrometer was used for the analysis of neurotransmitters in the brains of rats exposed to TBBPA in gestation and lactation through their mothers. Three neurotransmitters of interest were studied in three selected regions of the brain: caudate putamen, substantia nigra (SN), and dorsal raphe. Stable isotope labeled (SIL) standards were used as internal standards and a means to achieve relative quantification. This study serves as a demonstration of a new application of IR-MALDESI, namely that neurotransmitter distributions can be confidently and rapidly imaged without derivatization.


IR-MALDESI Neurotransmitters Mass spectrometry imaging Exposomics Orbitrap 



All mass spectrometry measurements were made in the Molecular Education, Technology, and Research Innovation Center (METRIC) at NC State University. We are grateful for the NIEHS research team who designed and executed the TBBPA exposure portion of this project, especially Suzanne Fenton, Manushree Bharadwaj, Joshua Warmack, and Sagi Enicole Gillera.

Funding information

This study received financial assistance from the National Institutes of Health grants R01GM087964 (MCB, ME, DM) and P30ES025128 to North Carolina State University as well as a National Institutes of Health IPA Agreement with HP.

Compliance with ethical standards

All aspects of the rat studies were approved by the Institutional Animal Care and Use Committees of NIEHS and NCSU.

Conflict of interest

The authors declare that they have no conflicts of interest

Supplementary material

216_2018_1420_MOESM1_ESM.pdf (3.4 mb)
ESM 1 (PDF 3448 kb)


  1. 1.
    Galanopoulou AS. Sexually dimorphic expression of KCC2 and GABA function. Epilepsy Res. 2008. Scholar
  2. 2.
    How many chemicals are in use today? C&EN Global Enterp. 2017.
  3. 3.
    Heyer DB, Meredith RM. Environmental toxicology: sensitive periods of development and neurodevelopmental disorders. Neurotoxicology. 2017.Google Scholar
  4. 4.
    Malliari E, Kalantzi O. Children’s exposure to brominated flame retardants in indoor environments - a review. Environ Int. 2017;108:146–69.CrossRefGoogle Scholar
  5. 5.
    Sugeng EJ, de Cock M, Schoonmade LJ, van de Bor M. Toddler exposure to flame retardant chemicals: magnitude, health concern and potential risk- or protective factors of exposure: observational studies summarized in a systematic review. Chemosphere. 2017;184:820–31.CrossRefGoogle Scholar
  6. 6.
    Dingemans MML, van dB, Westerink RHS. Neurotoxicity of brominated flame retardants: (in)direct effects of parent and hydroxylated polybrominated diphenyl ethers on the (developing) nervous system. Environ Health Perspect. 2011. Scholar
  7. 7.
    Stapleton HM, Sharma S, Getzinger G, Ferguson PL, Gabriel M, Webster TF, et al. Novel and high volume use flame retardants in US couches reflective of the 2005 PentaBDE phase out. Environ Sci Technol. 2012. Scholar
  8. 8.
    Knudsen GA, Hughes MF, McIntosh KL, Sanders JM, Birnbaum LS. Estimation of tetrabromobisphenol A (TBBPA) percutaneous uptake in humans using the parallelogram method. Toxicol Appl Pharmacol. 2015. Scholar
  9. 9.
    Antignac JP, Cariou R, Zalko D, Berrebi A, Cravedi JP, Maume D, et al. Exposure assessment of French women and their newborn to brominated flame retardants: determination of tri- to deca- polybromodiphenylethers (PBDE) in maternal adipose tissue, serum, breast milk and cord serum. Environ Pollut. 2009. Scholar
  10. 10.
    Cariou R, Antignac JP, Zalko D, Berrebi A, Cravedi JP, Maume D, et al. Exposure assessment of French women and their newborns to tetrabromobisphenol-A: occurrence measurements in maternal adipose tissue, serum, breast milk and cord serum. Chemosphere. 2008. Scholar
  11. 11.
    Lai DY, Kacew S, Dekant W. Tetrabromobisphenol A (TBBPA): possible modes of action of toxicity and carcinogenicity in rodents. Food Chem Toxicol. 2015. Scholar
  12. 12.
    National TP. NTP technical report on the toxicology studies of tetrabromobisphenol A (CASRN 79-94-7) in F344/NTac rats and B6C3F1/N mice and toxicology and carcinogenesis studies of tetrabromobisphenol A in Wistar Han Crl:WI(Han)] rats and B6C3F1/N mice (gavage studies). U.S. Department of Health and Human Services, Public Health Service, National Institutes of Health, National Toxicology Program. 2014.Google Scholar
  13. 13.
    Nakajima A, Saigusa D, Tetsu N, Yamakuni T, Tomioka Y, Hishinuma T. Neurobehavioral effects of tetrabromobisphenol A, a brominated flame retardant, in mice. Toxicol Lett. 2009. Scholar
  14. 14.
    Chughtai K, Heeren RMA. Mass spectrometric imaging for biomedical tissue analysis. Chem Rev. 2010. Scholar
  15. 15.
    Tanaka K, Waki H, Ido Y, Akita S, Yoshida Y, Yoshida T, et al. Protein and polymer analyses up to m/z 100 000 by laser ionization time-of-flight mass spectrometry. Rapid Commun Mass Spectrom. 1988. Scholar
  16. 16.
    Karas M, Hillenkamp F. Laser desorption ionization of proteins with molecular masses exceeding 10,000 daltons. Anal Chem. 1988. Scholar
  17. 17.
    Fenn JB, Mann M, Meng CK, Wong SF, Whitehouse CM. Electrospray ionization for mass spectrometry of large biomolecules. Science. 1989. Scholar
  18. 18.
    Norris JL, Caprioli RM. Analysis of tissue specimens by matrix-assisted laser desorption/ionization imaging mass spectrometry in biological and clinical research. Chem Rev. 2013. Scholar
  19. 19.
    Shariatgorji M, Nilsson A, Goodwin RJ, Kallback P, Schintu N, Zhang X, et al. Direct targeted quantitative molecular imaging of neurotransmitters in brain tissue sections. Neuron. 2014. Scholar
  20. 20.
    Nemes P, Woods AS, Vertes A. Simultaneous imaging of small metabolites and lipids in rat brain tissues at atmospheric pressure by laser ablation electrospray ionization mass spectrometry. Anal Chem. 2010. Scholar
  21. 21.
    Bergman HM, Lundin E, Andersson M, Lanekoff I. Quantitative mass spectrometry imaging of small-molecule neurotransmitters in rat brain tissue sections using nanospray desorption electrospray ionization. Analyst. 2016. Scholar
  22. 22.
    Fernandes AMAP, Vendramini PH, Galaverna R, Schwab NV, Alberici LC, Augusti R, et al. Direct visualization of neurotransmitters in rat brain slices by desorption electrospray ionization mass spectrometry imaging (DESI - MS). J Am Soc Mass Spectrom. 2016. Scholar
  23. 23.
    Passarelli MK, Winograd N. Lipid imaging with time-of-flight secondary ion mass spectrometry (ToF-SIMS). Biochim Biophys Acta. 2011. Scholar
  24. 24.
    Bokhart MT, Muddiman DC. Infrared matrix-assisted laser desorption electrospray ionization mass spectrometry imaging analysis of biospecimens. Analyst. 2016. Scholar
  25. 25.
    Robichaud G, Barry J, Muddiman D. IR-MALDESI mass spectrometry imaging of biological tissue sections using ice as a matrix. J Am Soc Mass Spectrom. 2014. Scholar
  26. 26.
    Esteve C, Tolner EA, Shyti R, van dM, McDonnell LA. Mass spectrometry imaging of amino neurotransmitters: a comparison of derivatization methods and application in mouse brain tissue. Metabolomics. 2016.
  27. 27.
    Sakino T, Yuki S, Akiko K, Mitsuyo O, Sachise K, Toshimi M, et al. Microscopic imaging mass spectrometry assisted by on-tissue chemical derivatization for visualizing multiple amino acids in human colon cancer xenografts. Proteomics. 2014. Scholar
  28. 28.
    Shariatgorji M, Nilsson A, Goodwin RA, Källback P, Schintu N, Zhang X, et al. Direct targeted quantitative molecular imaging of neurotransmitters in brain tissue sections. Neuron. 2014;84:697–707.CrossRefGoogle Scholar
  29. 29.
    Bokhart MT, Rosen E, Thompson C, Sykes C, Kashuba ADM, Muddiman DC. Quantitative mass spectrometry imaging of emtricitabine in cervical tissue model using infrared matrix-assisted laser desorption electrospray ionization. Anal Bioanal Chem. 2015. Scholar
  30. 30.
    Paxinos G, Watson C. The rat brain in stereotaxic coordinates. London: Academic Press; 2007.Google Scholar
  31. 31.
    Arambula SE, Fuchs J, Cao J, Patisaul HB. Effects of perinatal bisphenol A exposure on the volume of sexually-dimorphic nuclei of juvenile rats: a CLARITY-BPA consortium study. NeuroToxicology. 2017;63:33–42.CrossRefGoogle Scholar
  32. 32.
    Cao J, Joyner L, Mickens JA, Leyrer SM, Patisaul HB. Sex-specific Esr2 mRNA expression in the rat hypothalamus and amygdala is altered by neonatal bisphenol A exposure. Reproduction. 2014. Scholar
  33. 33.
    Ekelöf M, Manni J, Nazari M, Bokhart M, Muddiman DC. Characterization of a novel miniaturized burst-mode infrared laser system for IR-MALDESI mass spectrometry imaging. Anal Bioanal Chem. 2018. Scholar
  34. 34.
    Robichaud G, Garrard KP, Barry JA, Muddiman DC. MSiReader: an open-source interface to view and analyze high resolving power MS imaging files on Matlab platform. J Am Soc Mass Spectrom. 2013. Scholar
  35. 35.
    Bokhart MT, Nazari M, Garrard KP, Muddiman DC. MSiReader v1.0: evolving open-source mass spectrometry imaging software for targeted and untargeted analyses. J Am Soc Mass Spectrom. 2017. Scholar
  36. 36.
    Chambers MC, Maclean B, Burke R, Amodei D, Ruderman DL, Neumann S, et al. A cross-platform toolkit for mass spectrometry and proteomics. Nat Biotechnol. 2012. Scholar
  37. 37.
    Race AM, Styles IB, Bunch J. Inclusive sharing of mass spectrometry imaging data requires a converter for all. J Proteomics. 2012. Scholar
  38. 38.
    Hines M, Allen LS, Gorski RA. Sex differences in subregions of the medial nucleus of the amygdala and the bed nucleus of the stria terminalis of the rat. Brain Res. 1992;579:321–6.CrossRefGoogle Scholar
  39. 39.
    Frazer A, Hensler JG. Anonymous basic neurochemistry molecular, cellular and medical aspects. 6th ed. Philadelphia: Lippincott-Raven; 1999.Google Scholar
  40. 40.
    Yu X, Ye Z, Houston C, Zecharia A, Ma Y, Zhang Z, et al. Wakefulness is governed by GABA and histamine cotransmission. Neuron. 2015. Scholar
  41. 41.
    Ponvert C, Galoppin L, Scheinmann P, Canu P, Burtin C. Tissue histamine levels in male and female mast cell deficient mice (W/Wv) and in their littermates (Wv/+, W/+ and +/+). Agents Actions. 1985. Scholar
  42. 42.
    Netter KJ, Cohn VH, Shore PA. Sex difference in histamine metabolism in the rat. Am J Physiol. 1961. Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • M. Caleb Bagley
    • 1
  • Måns Ekelöf
    • 1
  • Kylie Rock
    • 2
    • 3
  • Heather Patisaul
    • 2
    • 3
  • David C. Muddiman
    • 1
    • 3
    • 4
    Email author
  1. 1.FTMS Laboratory for Human Health Research, Department of ChemistryNorth Carolina State UniversityRaleighUSA
  2. 2.Department of Biological SciencesNorth Carolina State UniversityRaleighUSA
  3. 3.Center for Human Health and the EnvironmentNorth Carolina State UniversityRaleighUSA
  4. 4.Molecular Education, Technology and Research Innovation Center (METRIC)North Carolina State UniversityRaleighUSA

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