Abstract
Introduction
In-situ detection and in particular comprehensive analysis of small molecule metabolites (SMMs, m/z < 500) using matrix-assisted laser desorption ionization mass spectrometry imaging (MALDI MSI) remain a challenge, mainly due to ion suppression effects from more abundant molecules in tissue section like lipids.
Objective
A strategy based on organic washes to remove most ionization-suppressing lipids from tissue section was firstly explored for improved analysis of SMMs by MALDI MSI.
Methods
The tissue sections after rinse with different organic solvents were analyzed by MALDI MSI, and the results were compared for the optimized washing conditions.
Results
The rinse with chloroform for 15 s at − 20 °C significantly removed most glycerophospholipids and glycerolipids from tissue section. Consequentially, ATP-related energy metabolites, amino acids and derivatives, glucose derivatives, glycolysis pathway metabolites and other SMMs were able to be well-visualized with enhanced ion intensity and good reproducibility. The organic washes-based MALDI MSI was applied to the metabolic pathway analysis in rat brain following status epilepticus (SE) model, which was, as far as we know, the first report about in-situ detection of a broad range of metabolites in the model of SE by MALDI MSI technique. The alterations of cyclic adenosine monophosphate (cyclic AMP), inosine, glutamine, glutathione, taurine and spermine during SE were observed.
Conclusion
A simple organic washing protocol enables comprehensive analysis of tissue SMMs in MALDI MSI by removing ionization-suppressing lipids. The application in the SE model indicates that MALDI MSI analysis potentially provides new insight for understanding the disease mechanism.
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References
Alvestad, S., Hammer, J., Qu, H., Haberg, A., Ottersen, O. P., & Sonnewald, U. (2011). Reduced astrocytic contribution to the turnover of glutamate, glutamine, and GABA characterizes the latent phase in the kainate model of temporal lobe epilepsy. Journal of Cerebral Blood Flow and Metabolism, 31(8), 1675–1686. https://doi.org/10.1038/jcbfm.2011.36.
Angel, P. M., Spraggins, J. M., Baldwin, H. S., & Caprioli, R. (2012). Enhanced sensitivity for high spatial resolution lipid analysis by negative ion mode matrix assisted laser desorption ionization imaging mass spectrometry. Analytical Chemistry, 84(3), 1557–1564. https://doi.org/10.1021/ac202383m.
Calligaris, D., Longuespee, R., Debois, D., Asakawa, D., Turtoi, A., Castronovo, V., et al. (2013). Selected protein monitoring in histological sections by targeted MALDI-FTICR in-source decay imaging. Analytical Chemistry, 85(4), 2117–2126. https://doi.org/10.1021/ac302746t.
Dona, F., Conceicao, I. M., Ulrich, H., Ribeiro, E. B., Freitas, T. A., Abrahao Nencioni, A. L., et al. (2016). Variations of ATP and its metabolites in the hippocampus of rats subjected to pilocarpine-induced temporal lobe epilepsy. Purinergic Signalling, 12(2), 295–302. https://doi.org/10.1007/s11302-016-9504-9.
Eid, T., Thomas, M. J., Spencer, D. D., Runden-Pran, E., Lai, J. C. K., Malthankar, G. V., et al. (2004). Loss of glutamine synthetase in the human epileptogenic hippocampus: Possible mechanism for raised extracellular glutamate in mesial temporal lobe epilepsy. Lancet, 363(9402), 28–37. https://doi.org/10.1016/s0140-6736(03)15166-5.
Esteve, C., Tolner, E. A., Shyti, R., van den Maagdenberg, A. M. J. M., & McDonnell, L. A. (2016). Mass spectrometry imaging of amino neurotransmitters: A comparison of derivatization methods and application in mouse brain tissue. Metabolomics https://doi.org/10.1007/s11306-015-0926-0.
Filibian, M., Frasca, A., Maggioni, D., Micotti, E., Vezzani, A., & Ravizza, T. (2012). In vivo imaging of glia activation using H-1-magnetic resonance spectroscopy to detect putative biomarkers of tissue epileptogenicity. Epilepsia, 53(11), 1907–1916. https://doi.org/10.1111/j.1528-1167.2012.03685.x.
Folch, J., Lees, M., & SLOANE STANLEY, G. H. (1957). A simple method for the isolation and purification of total lipids from animal tissue. Journal of Biological Chemistry, 226(1), 497–509.
Gatenby, R. A., & Gillies, R. J. (2004). Why do cancers have high aerobic glycolysis? Nature Reviews Cancer, 4(11), 891–899. https://doi.org/10.1038/nrc1478.
Green, D. R., Galluzzi, L., & Kroemer, G. (2014). Metabolic control of cell death. Science, 345(6203), 1466–1490. https://doi.org/10.1126/science.1250256.
Guo, S., Wang, Y., Zhou, D., & Li, Z. (2015). Electric field-assisted matrix coating method enhances the detection of small molecule metabolites for mass spectrometry imaging. Analytical Chemistry, 87(12), 5860–5865. https://doi.org/10.1021/ac504761t.
Guo, Z., Zhang, Q. C., Zou, H. F., Guo, B. C., & Ni, J. Y. (2002). A method for the analysis of low-mass molecules by MALDI-TOF mass spectrometry. Analytical Chemistry, 74(7), 1637–1641. https://doi.org/10.1021/ac010979m.
Harmsen, E., Detombe, P. P., Dejong, J. W., & Achterberg, P. W. (1984). Enhanced ATP and GTP synthesis from hypoxanthine or inosine after myocardial ischemia. American Journal of Physiology, 246(1), H37–H43.
Hillert, M. H., Imran, I., Zimmermann, M., Lau, H., Weinfurter, S., & Klein, J. (2014). Dynamics of hippocampal acetylcholine release during lithium-pilocarpine-induced status epilepticus in rats. Journal of Neurochemistry, 131(1), 42–52. https://doi.org/10.1111/jnc.12787.
Hua, Y. M., Dagan, S., Wickramasekara, S., Boday, D. J., & Wysocki, V. H. (2010). Analysis of deprotonated acids with silicon nanoparticle-assisted laser desorption/ionization mass spectrometry. Journal of Mass Spectrometry, 45(12), 1394–1401. https://doi.org/10.1002/jms.1852.
Karlsson, O., & Hanrieder, J. (2016). Imaging mass spectrometry in drug development and toxicology. Archives of Toxicology, 91(6), 2283–2294.
Kovacs, Z., Kekesi, K. A., Juhasz, G., Barna, J., Heja, L., Lakatos, R., et al. (2015). Non-adenosine nucleoside inosine, guanosine and uridine as promising antiepileptic drugs: a summary of current literature. [Research Support, Non-U.S. Gov’t; Review]. Mini Reviews in Medicinal Chemistry, 14(13), 1033–1042.
Kovacs, Z., Kekesi, K. A., Juhasz, G., & Dobolyi, A. (2014). The antiepileptic potential of nucleosides. Current Medicinal Chemistry, 21(6), 788–821.
Lemaire, R., Wisztorski, M., Desmons, A., Tabet, J. C., Day, R., Salzet, M., et al. (2006). MALDI-MS direct tissue analysis of proteins: Improving signal sensitivity using organic treatments. Analytical Chemistry, 78(20), 7145–7153. https://doi.org/10.1021/ac060565z.
Li, S., Zhang, Y., Liu, J. a., Han, J., Guan, M., Yang, H., et al. (2016). Electrospray deposition device used to precisely control the matrix crystal to improve the performance of MALDI MSI. Scientific Reports, 6, 37903. https://doi.org/10.1038/srep37903.
Lietsche, J., Imran, I., & Klein, J. (2016). Extracellular levels of ATP and acetylcholine during lithium-pilocarpine induced status epilepticus in rats. Neuroscience Letters, 611, 69–73. https://doi.org/10.1016/j.neulet.2015.11.028.
Liu, H., Chen, R., Wang, J., Chen, S., Xiong, C., Wang, J., et al. (2014). 1,5-Diaminonaphthalene hydrochloride assisted laser desorption/ionization mass spectrometry imaging of small molecules in tissues following focal cerebral ischemia. Analytical Chemistry, 86(20), 10114–10121. https://doi.org/10.1021/ac5034566.
Meng, J., Shi, C., & Deng, C. (2011). Facile synthesis of water-soluble multi-wall carbon nanotubes and polyaniline composites and their application in detection of small metabolites by matrix assisted laser desorption/ionization mass spectrometry. Chemical Communications, 47(39), 11017–11019. https://doi.org/10.1039/c1cc14319k.
Miura, D., Fujimura, Y., Tachibana, H., & Wariishi, H. (2010a). Highly sensitive matrix-assisted laser desorption ionization-mass spectrometry for high-throughput metabolic profiling. Analytical Chemistry, 82(2), 498–504. https://doi.org/10.1021/ac901083a.
Miura, D., Fujimura, Y., Yamato, M., Hyodo, F., Utsumi, H., Tachibana, H., et al. (2010b). Ultrahighly sensitive in situ metabolomic imaging for visualizing spatiotemporal metabolic behaviors. Analytical Chemistry, 82(23), 9789–9796. https://doi.org/10.1021/ac101998z.
Norris, J. L., & Caprioli, R. M. (2013). Analysis of tissue specimens by matrix-assisted laser desorption/ionization imaging mass spectrometry in biological and clinical research. Chemical Reviews, 113(4), 2309–2342. https://doi.org/10.1021/cr3004295.
Nugent, A. C., Martinez, A., D’Alfonso, A., Zarate, C. A., & Theodore, W. H. (2015). The relationship between glucose metabolism, resting-state fMRI BOLD signal, and GABA(A)-binding potential: A preliminary study in healthy subjects and those with temporal lobe epilepsy. Journal of Cerebral Blood Flow and Metabolism, 35(4), 583–591. https://doi.org/10.1038/jcbfm.2014.228.
Padma, M. V., Simkins, R., White, P., Satter, M., Christian, B. T., Dunigan, K., et al. (2004). Clinical utility of 11C-flumazenil positron emission tomography in intractable temporal lobe epilepsy. Neurology India, 52(4), 457–462.
Patti, G. J., Yanes, O., & Siuzdak, G. (2012). Metabolomics: The apogee of the omics trilogy. Nature Reviews Molecular Cell Biology, 13(4), 263–269. https://doi.org/10.1038/nrm3314.
Paxinos, G., & Watson, C. (1986). The rat brain in stereotaxic coordinates. San Diego: Academic Press.
Racine, R., Okujava, V., & Chipashvili, S. (1972). Modification of seizure activity by electrical stimulation: III. Mechanisms. Electroencephalography & Clinical Neurophysiology, 32(3), 295–299.
Rice, A. C., & DeLorenzo, R. J. (1998). NMDA receptor activation during status epilepticus is required for the development of epilepsy. Brain Research, 782(1–2), 240–247. https://doi.org/10.1016/s0006-8993(97)01285-7.
Shi, C. Y., & Deng, C. H. (2016). Recent advances in inorganic materials for LDI-MS analysis of small molecules. Analyst, 141(10), 2816–2826. https://doi.org/10.1039/c6an00220j.
Smeland, O. B., Hadera, M. G., McDonald, T. S., Sonnewald, U., & Borges, K. (2013). Brain mitochondrial metabolic dysfunction and glutamate level reduction in the pilocarpine model of temporal lobe epilepsy in mice. Journal of Cerebral Blood Flow and Metabolism, 33(7), 1090–1097. https://doi.org/10.1038/jcbfm.2013.54.
Sumner, L. W., Amberg, A., Barrett, D., Beale, M. H., Beger, R., Daykin, C. A., et al. (2007). Proposed minimum reporting standards for chemical analysis. Metabolomics, 3(3), 211–221. https://doi.org/10.1007/s11306-007-0082-2.
Thomas, A., Lenglet, S., Chaurand, P., Deglon, J., Mangin, P., Mach, F., et al. (2011). Mass spectrometry for the evaluation of cardiovascular diseases based on proteomics and lipidomics. Thrombosis and Haemostasis, 106(1), 20–33. https://doi.org/10.1160/th10-12-0812.
Thomas, A., Patterson, N. H., Charbonneau, J. L., & Chaurand, P. (2013). Orthogonal organic and aqueous-based washes of tissue sections to enhance protein sensitivity by MALDI imaging mass spectrometry. Journal of Mass Spectrometry, 48(1), 42–48. https://doi.org/10.1002/jms.3114.
van der Hel, W. S., Notenboom, R. G. E., Bos, I. W. M., van Rijen, P. C., van Veelen, C. W. M., & de Graan, P. N. E (2005). Reduced glutamine synthetase in hippocampal areas with neuron loss in temporal lobe epilepsy. Neurology, 64(2), 326–333.
Vens-Cappell, S., Kouzel, I. U., Kettling, H., Soltwisch, J., Bauwens, A., Porubsky, S., et al. (2016). On-tissue phospholipase C digestion for enhanced MALDI-MS imaging of neutral glycosphingolipids. Analytical Chemistry, 88(11), 5595–5599. https://doi.org/10.1021/acs.analchem.6b01084.
Wang, C., Xie, N., Wang, Y., Li, Y., Ge, X., & Wang, M. (2015). Role of the mitochondrial calcium uniporter in rat hippocampal neuronal death after pilocarpine-induced status epilepticus. Neurochemical Research, 40(8), 1739–1746. https://doi.org/10.1007/s11064-015-1657-3.
Wishart, D. S., Jewison, T., Guo, A. C., Wilson, M., Knox, C., Liu, Y., et al. (2013). HMDB 3.0-The human metabolome database in 2013. Nucleic Acids Research, 41(D1), D801–D807. https://doi.org/10.1093/nar/gks1065.
Wishart, D. S., Knox, C., Guo, A. C., Eisner, R., Young, N., Gautam, B., et al. (2009). HMDB: A knowledgebase for the human metabolome. Nucleic Acids Research, 37, D603–D610. https://doi.org/10.1093/nar/gkn810.
Wishart, D. S., Tzur, D., Knox, C., Eisner, R., Guo, A. C., Young, N., et al. (2007). HMDB: The human metabolome database. Nucleic Acids Research, 35, D521–D526. https://doi.org/10.1093/nar/gkl923.
Wu, Q., Comi, T. J., Li, B., Rubakhin, S. S., & Sweedler, J. V. (2016). On-tissue derivatization via electrospray deposition for matrix assisted laser desorption/ionization mass spectrometry imaging of endogenous fatty acids in rat brain tissues. Analytical Chemistry, 88(11), 5988–5995. https://doi.org/10.1021/acs.analchem.6b01021.
Wu, Y., Pearce, P. S., Rapuano, A., Hitchens, T. K., de Lanerolle, N. C., & Pan, J. W. (2015). Metabolic changes in early poststatus epilepticus measured by MR spectroscopy in rats. Journal of Cerebral Blood Flow and Metabolism, 35(11), 1862–1870. https://doi.org/10.1038/jcbfm.2015.145.
Xie, X. Y., Jiang, Y. C., Yuan, Y., Wang, P. Q., Li, X. Y., Chen, F. M., et al. (2016). MALDI imaging reveals NCOA7 as a potential biomarker in oral squamous cell carcinoma arising from oral submucous fibrosis. Oncotarget, 7(37), 59987–60004. https://doi.org/10.18632/oncotarget.11046.
Xu, G., Liu, S., Peng, J., Lv, W., & Wu, R. a. (2015). Facile synthesis of gold@graphitized mesoporous silica nanocomposite and its surface-assisted laser desorption/lonization for time-of-flight mass spectroscopy. ACS Applied Materials & Interfaces, 7(3), 2032–2038. https://doi.org/10.1021/am507894y.
Yaari, Y., Yue, C., Su, H.. Yaari, Y., Yue, C., & Su, H. (2007). Recruitment of apical dendritic T-type Ca2+ channels by backpropagating spikes underlies de novo intrinsic bursting in hippocampal epileptogenesis. Journal of Physiology, 580(Pt. 2), 435–450.
Yagnik, G. B., Hansen, R. L., Korte, A. R., Reichert, M. D., Vela, J., & Lee, Y. J. (2016). Large scale nanoparticle screening for small molecule analysis in laser desorption ionization mass spectrometry. Analytical Chemistry, 88(18), 8926–8930. https://doi.org/10.1021/acs.analchem.6b02732.
Yang, X., Lin, Z., Yan, X., & Cai, Z. (2016). Zeolitic imidazolate framework nanocrystals for enrichment and direct detection of environmental pollutants by negative ion surface-assisted laser desorption/ionization time-of-flight mass spectrometry. RSC Advances, 6(28), 23790–23793. https://doi.org/10.1039/c6ra00877a.
Yuan, Y., Zhou, H., & Wei, Y. (2005). Effects of melatonin on contents of hippocampal cyclic AMP in rats witH L-glutamate-induced seizures. Chinese Journal of Histochemistry and Cytochemistry, 14(3), 331–334.
Acknowledgements
This work was supported by National Natural Science Foundation of China (Grant Nos. 21575146, 21635008 and 21621062).
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The animal experiments were performed according to “the Guide for the Care and Use of Laboratory Animals” from the Association for Assessment and Accreditation of Laboratory Animal Care and ethical approval of the present study was obtained from the Animal Care and Use Committee at National Center for Nanoscience and Technology of China.
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Yang, H., Ji, W., Guan, M. et al. Organic washes of tissue sections for comprehensive analysis of small molecule metabolites by MALDI MS imaging of rat brain following status epilepticus. Metabolomics 14, 50 (2018). https://doi.org/10.1007/s11306-018-1348-6
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DOI: https://doi.org/10.1007/s11306-018-1348-6