HPLC-ESI-MS determination of neurotransmitters L-serine/D-serine in rat brain tissues

Abstract

Abstract: A highly sensitive and selective high performance liquid chromatography-electrospray ionization-mass spectrometry (HPLC-ESI-MS) assay was developed for the determination of neurotransmitters L-serine/D-serine in rat brain tissues. The rat brain was removed, homogenized and centrifuged.L-serine/D-serine in the supernatant was derivatized with the Marfey’s reagent. The derivatives were separated by HPLC under gradient elution of mobile phase consisting of ammonium formate and acetonitrile and detected by ESI-MS in the negative ion mode. Reverse phase chromatographic resolution of the enantiomers was obtained within 22 min. Good linear correlations of the concentration to the peak area were obtained in the ranges of 2.5~500 ng/mg for L-serine and 1.2~250 ng/mg for D-serine. The extraction recoveries for both analysts were in the range of 81% to 89%. The method was then applied to determine the levels of the neurotransmitters in cerebral cortex and hippocampus of the adult and aged rats. The results showed that L-serine levels kept unchanged,while D-serine levels decreased significantly, in cortex and hippocampus of the aged rats compared with those of the adult rats, suggesting an involvement of D-serine in the normal aging process.

Key words: HPLC-ESI-MS, brain tissue, neurotransmitter, D-serine, L-serine

CLC Number:

O657.63

WANG Wei-li, DUAN Ya-le, LIU Sha, ZHANG Yao, ZHAO Zheng. HPLC-ESI-MS determination of neurotransmitters L-serine/D-serine in rat brain tissues[J]. Journal of East China Normal University(Natural Sc, 2013, 2013(6): 165-170.

References

［1］ CAVALHEIRO E A, FERNANDES M J, TURSKI L, et al. Spontaneous recurrent seizures in rats: amino acid and monoamine determination in the hippocampus［J］. Epilepsia, 1994, 35(1): 1-11.

［2］ MULDNER A, HOYER S. Delayed decrease of excitatory amino acid neurotransmitters in parietotemporal cortex and hippocampus after complete cerebral ischemia in aged rats［J］. Archives of Gerontology and Geriatrics, 1997, 24(1): 23-33.

［3］ IMAI K, FUKUSHIMA T, SANTA T, et al. Distribution of free D-amino acids in tissues and body fluids of vertebrates［J］. Enantiomer, 1997, 2(3-4): 143-5.

［4］ SCHELL M J. The N-methyl D-aspartate receptor glycine site and D-serine metabolism: an evolutionary perspective［J］. Philosophical Transactions of the Royal Society of London Series B-Biological Sciences, 2004, 359(1446): 943-964.

［5］ BILLARD J M. D-serine signalling as a prominent determinant of neuronal-glial dialogue in the healthy and diseased brain［J］. J Cell Mol Med, 2008, 12(5B): 1872-84.

［6］ NAGAI T, YU J, KITAHARA Y, et al. D-serine ameliorates neonatal polyI: C treatment-induced emotional and cognitive impairments in adult mice［J］. Journal of Pharmacological Sciences, 2012, 120(3): 213-27.

［7］ FOSSAT P, TURPIN F R, SACCHI S, et al. Glial D-serine gates NMDA receptors at excitatory synapses in prefrontal cortex［J］. Cerebral Cortex, 2012, 22(3): 595-606.

［8］ BILLARD J M. D-serine signalling as a prominent determinant of neuronal-glial dialogue in the healthy and diseased brain［J］. Journal of Cellular and Molecular Medicine, 2008, 12(5B): 1872-1884.

［9］ HASHIMOTO K, FUKUSHIMA T, SHIMIZU E, et al. Possible role of d-serine in the pathophysiology of Alzheimer's disease［J］. Progress in Neuro-Psychopharmacology and Biological Psychiatry, 2004, 28(2): 385-388.

［10］ ABOU EL-MAGD R M, PARK H K, KAWAZOE T, et al. The effect of risperidone on D-amino acid oxidase activity as a hypothesis for a novel mechanism of action in the treatment of schizophrenia［J］. Journal of Psychopharmacology, 2010, 24(7): 1055-67.

［11］ SETHURAMAN R, KRISHNAMOORTHY M G, LEE T L, et al. Simultaneous analysis of D- and L-serine in cerebrospinal fluid by use of HPLC［J］. Clinical Chemistry, 2007, 53(8): 1489-94.

［12］ PAPPA-LOUISI A, AGRAFIOTOU P, SOTIROPOULOS S. Optimal conditions for the direct RP-HPLC determination of underivatized amino acids with online multiple detection［J］. Methods Mol Biol, 2012, 828: 101-14.

［13］ GU J, LI X J, SUN F Y. Melatonin decreases the release of amino acid transmitters in hypoxic rat cortical slice［J］. Acta Physiologica Sinica, 1999, 51(4): 445-8.

［14］ HASHIMOTO A, KUMASHIRO S, NISHIKAWA T, et al. Embryonic development and postnatal changes in free D-aspartate and D-serine in the human prefrontal cortex［J］. J Neurochem, 1993, 61(1): 348-51.

［15］ TURPIN F R, POTIER B, DULONG J R, et al. Reduced serine racemase expression contributes to age-related deficits in hippocampal cognitive function［J］. Neurobiol Aging, 2011, 32(8): 1495-504.

［16］ HAXAIRE C, TURPIN F R, POTIER B, et al. Reversal of age-related oxidative stress prevents hippocampal synaptic plasticity deficits by protecting D-serine-dependent NMDA receptor activation［J］. Aging Cell, 2012, 11(2): 336-44.

［17］ YANG Y, GE W, CHEN Y, et al. Contribution of astrocytes to hippocampal long-term potentiation through release of D-serine［J］. Proc Natl Acad Sci USA, 2003, 100(25):15194-5199.

［18］ YANG S, QIAO H, WEN L, et al. D-serine enhances impaired long-term potentiation in CA1 subfield of hippocampal slices from aged senescence-accelerated mouse prone/8 ［J］. Neurosci Lett, 2005, 379(1):7-12.