Determination of free d-aspartic acid,d-serine and d-alanine in the brain of mutant mice lacking d-amino-acid oxidase activity |
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| Affiliation: | 1. Translational Neuroscience Unit, IRCCS Casa Sollievo della Sofferenza, 71013, San Giovanni Rotondo, Italy;2. Laboratory of Neuronal Cell Signaling, EBRI Rita Levi-Montalcini Foundation, Rome, Italy;3. Neurobiology in Translational Medicine, Department of Neurorehabilitation Sciences, Casa Cura Policlinico, Milan, Italy;4. Institute of Biomolecular Chemistry (I.C.B.), National Research Council (C.N.R.), 80078, Pozzuoli, Naples, Italy;5. Institute of Cell Biology and Neurobiology, 00015, Monterotondo Scalo, Rome, Italy;6. Department of Sciences and Technologies, University of Sannio, 82100, Benevento, Italy;7. Department of Veterinary Medicine and Animal Productions, University of Naples Federico II, Naples, Italy;8. Stazione Zoologica Anton Dohrn, Naples, Italy;9. Institute of Cell Biology and Neurobiology/CNR, Department of Sense Organs, Sapienza University of Rome, Italy;10. Department of Biotechnology and Life Sciences, University of Insubria, 21100, Varese, Italy;11. Department of Environmental, Biological and Pharmaceutical Sciences and Technologies, University of Campania Luigi Vanvitelli, 81100, Caserta, Italy;12. Laboratory of Behavioural Neuroscience, Ceinge Biotecnologie Avanzate, 80145, Naples, Italy;13. Department of Agricultural Sciences, University of Naples “Federico II”, 80055, Portici, Italy;14. Department of System Medicine, University of Rome “Tor Vergata”, 00137, Rome, Italy;1. Dipartimento di Biotecnologie e Scienze della Vita, Università degli studi dell''Insubria, via J. H. Dunant 3, 21100 Varese, Italy;2. The Protein Factory, Politecnico di Milano and Università degli studi dell''Insubria, via Mancinelli 7, 20131 Milan, Italy |
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| Abstract: | A simple and precise method for the simultaneous determination of free d-aspartic acid, d-serine and d-alanine in mouse brain tissues was established, using a reversed-phase HPLC system with widely used pre-column derivatizing reagents, o-phthaldialdehyde and N-t-butyloxycarbonyl-l-cysteine. With the present method, the contents of these three d-amino acids in hippocampus, hypothalamus, pituitary gland, pineal gland and medulla oblongata as well as cerebrum and cerebellum of mutant mice lacking d-amino-acid oxidase activity were determined and compared with those obtained for control mice. In both mice, extremely high contents of d-serine were observed in forebrain (100–400 nmol/g wet tissue), and the contents were small in pituitary and pineal glands. While, d-serine contents in cerebellum and medulla oblongata of mutant mice were about ten times higher than those in control mice. In contrast, d-alanine contents in mutant mice were higher than those in control mice in all brain regions and serum. |
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