Abstract
There is a growing interest in biological models to investigate the effect of neurotransmitter dysregulation on the structure and function of the central nervous system (CNS) at different stages of development. Zebrafish, a vertebrate model increasingly used in neurobiology and neurotoxicology, shares the common neurotransmitter systems with mammals, including glutamate, GABA, glycine, dopamine, norepinephrine, epinephrine, serotonin, acetylcholine, and histamine. In this study, we have evaluated the performance of liquid chromatography-tandem mass spectrometry (LC-MS/MS) for the multiresidue determination of neurotransmitters and related metabolites. In a first step, ionization conditions were tested in positive electrospray mode and optimum fragmentation patterns were determined to optimize two selected reaction monitoring (SRM) transitions. Chromatographic conditions were optimized considering the chemical structure and chromatographic behavior of the analyzed compounds. The best performance was obtained with a Synergy Polar-RP column, which allowed the separation of the 38 compounds in 30 min. In addition, the performance of LC-MS/MS was studied in terms of linearity, sensitivity, intra- and inter-day precision, and overall robustness. The developed analytical method was able to quantify 27 of these neurochemicals in zebrafish chemical models for mild (P1), moderate (P2), and severe (P3) acute organophosphorus poisoning (OPP). The results show a general depression of synaptic-related neurochemicals, including the excitatory and inhibitory amino acids, as well as altered phospholipid metabolism, with specific neurochemical profiles associated to the different grades of severity. These results confirmed that the developed analytical method is a new tool for neurotoxicology research using the zebrafish model.
Similar content being viewed by others
References
Horzmann KA, Freeman JL. Zebrafish get connected: investigating neurotransmission targets and alterations in chemical toxicity. Toxics. 2016;4(3):19.
Kurian MA, Gissen P, Smith M, Heales SJ, Clayton PT. The monoamine neurotransmitter disorders: an expanding range of neurological syndromes. Lancet Neurol. 2011;10(8):721–33.
Ng J, Papandreou A, Heales SJ, Kurian MA. Monoamine neurotransmitter disorders – clinical advances and future perspectives. Nat Rev Neurol. 2015;11(10):567–84.
Marecos C, Ng J, Kurian MA. What is new for monoamine neurotransmitter disorders? J Inherited Metab Dis. 2014;37(4):619–26.
Pearl PL, Hartka TR, Taylor J. Diagnosis and treatment of neurotransmitter disorders. Curr Treat Opt Neurol. 2006;8(6):441–50.
Pearl PL, Capp PK, Novotny EJ, Gibson KM. Inherited disorders of neurotransmitters in children and adults. Clin Biochem. 2005;38(12):1051–8.
Babin PJ, Goizet C, Raldúa D. Zebrafish models of human motor neuron diseases: advantages and limitations. Prog Neurobiol. 2014;118:36–58.
Faria M, Prats E, Padrós F, Soares AM, Raldúa D. Zebrafish is a predictive model for identifying compounds that protect against brain toxicity in severe acute organophosphorus intoxication. Arch Toxicol. 2016:1–11.
Raldúa D, Piña B. In vivo zebrafish assays for analyzing drug toxicity. Expert Opin Drug Metab Toxicol. 2014;10(5):685–97.
Tingaud-Sequeira A, Raldúa D, Lavie J, Mathieu G, Bordier M, Knoll-Gellida A, et al. Functional validation of ABHD12 mutations in the neurodegenerative disease PHARC. Neurobiol Dis. 2017;98:36–51.
Rico E, Rosemberg D, Seibt K, Capiotti K, Da Silva R, Bonan C. Zebrafish neurotransmitter systems as potential pharmacological and toxicological targets. Neurotoxicol Teratol. 2011;33(6):608–17.
Panula P, Sallinen V, Sundvik M, Kolehmainen J, Torkko V, Tiittula A, et al. Modulatory neurotransmitter systems and behavior: towards zebrafish models of neurodegenerative diseases. Zebrafish. 2006;3(2):235–47.
Sallinen V, Sundvik M, Reenilä I, Peitsaro N, Khrustalyov D, Anichtchik O, et al. Hyperserotonergic phenotype after monoamine oxidase inhibition in larval zebrafish. J Neurochem. 2009;109(2):403–15.
Tran S, Nowicki M, Muraleetharan A, Chatterjee D, Gerlai R. Neurochemical factors underlying individual differences in locomotor activity and anxiety-like behavioral responses in zebrafish. Prog Neuro-Psychopharmacol Biol Psychiat. 2016;65:25–33.
Tufi S, Leonards P, Lamoree M, De Boer J, Legler J, Legradi J. Changes in neurotransmitter profiles during early zebrafish (Danio rerio) development and after pesticide exposure. Environ Sci Technol. 2016;50(6):3222–30.
Højer-Pedersen J, Smedsgaard J, Nielsen J. The yeast metabolome addressed by electrospray ionization mass spectrometry: initiation of a mass spectral library and its applications for metabolic footprinting by direct infusion mass spectrometry. Metabolomics. 2008;4(4):393–405.
Bajad SU, Lu W, Kimball EH, Yuan J, Peterson C, Rabinowitz JD. Separation and quantitation of water soluble cellular metabolites by hydrophilic interaction chromatography-tandem mass spectrometry. J Chromatogr A. 2006;1125(1):76–88.
Gómez-Canela C, Miller TH, Bury NR, Tauler R, Barron LP. Targeted metabolomics of Gammarus pulex following controlled exposures to selected pharmaceuticals in water. Sci Total Environ. 2016;562:777–88.
Gómez-Canela C, Prats E, Piña B, Tauler R. Assessment of chlorpyrifos toxic effects in zebrafish (Danio rerio) metabolism. Environ Pollut. 2017;220:1231–43.
Lu H, Liang Y, Dunn WB, Shen H, Kell DB. Comparative evaluation of software for deconvolution of metabolomics data based on GC-TOF-MS. TrAC Trends Anal Chem. 2008;27(3):215–27.
Asiago VM, Alvarado LZ, Shanaiah N, Gowda GAN, Owusu-Sarfo K, Ballas RA, et al. Early detection of recurrent breast cancer using metabolite profiling. Cancer Res. 2010;70(21):8309–18.
Santos-Fandila A, Vázquez E, Barranco A, Zafra-Gómez A, Navalón A, Rueda R, et al. Analysis of 17 neurotransmitters, metabolites, and precursors in zebrafish through the life cycle using ultrahigh performance liquid chromatography-tandem mass spectrometry. J Chromatogr B: Anal Technol Biomed Life Sci. 2015;1001:191–201.
Aragon A, Legradi J, Ballesteros-Gómez A, Legler J, van Velzen M, de Boer J, et al. Determination of monoamine neurotransmitters in zebrafish (Danio rerio) by gas chromatography coupled to mass spectrometry with a two-step derivatization. Anal Bioanal Chem. 2017;409(11):2931–9.
Nusslein-Volhard C, Dahm R. Zebrafish: a practical approach. New York: Oxford University Press; 2002. p. 303.
Hildebrand DGC, Cicconet M, Torres RM, Choi W, Quan TM, Moon J, et al. Whole-brain serial-section electron microscopy in larval zebrafish. Nature. 2017;545(7654):345–9.
Faria M, Garcia-Reyero N, Padrós F, Babin PJ, Sebastián D, Cachot J, et al. Zebrafish Models for human acute organophosphorus poisoning. Sci Rep. 2015;5 https://doi.org/10.1038/srep15591.
Core Team R. A language and environment for statistical computing. Vienna: R Foundation for Statistical Computing; 2015.
Official Journal of the European Communities O (2002) Implementing Council Directive 96/23/EC concerning the performance of analytical methods and the interpretation of results. Directive 2002/657/CE.
Alpert AJ. Hydrophilic-interaction chromatography for the separation of peptides, nucleic acids and other polar compounds. J Chromatogr. 1990;499:177–96.
Tufi S, Lamoree M, de Boer J, Leonards P. Simultaneous analysis of multiple neurotransmitters by hydrophilic interaction liquid chromatography coupled to tandem mass spectrometry. J Chromatogr A. 2015;1395:79–87.
Stahnke H, Reemtsma T, Alder L. Compensation of matrix effects by postcolumn infusion of a monitor substance in multiresidue analysis with LC−MS/MS. Anal Chem. 2009;81(6):2185–92.
Shih T-M, McDonough JH. Neurochemical mechanisms in soman-induced seizures. J Appl Toxicol. 1997;17:255–64.
Rezaei M, Sadeghian A, Roohi N, Shojaei A, Mirnajafi-Zadeh J. Epilepsy and dopaminergic system. Physiol Pharmacol. 2017;21(1):1–14.
Lallement G, Denoyer M, Collet A, Pernot-Marino I, Baubichon D, Monmaur P, et al. Changes in hippocampal acetylcholine and glutamate extracellular levels during soman-induced seizures: influence of septal cholinoceptive cells. Neurosci Lett. 1992;139(1):104–7.
Acknowledgments
The research leading to these results has received funding from the European Research Council under European Union's Seven Framework Programme (FP/2007e2013)/ERC Grant Agreement n.320737, the NATO SfP project MD.SFPP 984777, and the grant CTM 2014-51985 from the Spanish Ministry of Economy, Industry and Competitiveness. Moreover, Rita Bausano is acknowledged for extraction samples support during her Erasmus Traineeship stage in the Environmental Chemistry Department at IDEA-CSIC (Barcelona, Spain).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare no financial conflict of interest.
Electronic supplementary material
ESM 1
(PDF 1.78 mb)
Rights and permissions
About this article
Cite this article
Gómez-Canela, C., Tornero-Cañadas, D., Prats, E. et al. Comprehensive characterization of neurochemicals in three zebrafish chemical models of human acute organophosphorus poisoning using liquid chromatography-tandem mass spectrometry. Anal Bioanal Chem 410, 1735–1748 (2018). https://doi.org/10.1007/s00216-017-0827-3
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00216-017-0827-3