title: Monitoring of the velocity of high-affinity glutamate uptake by isolated brain nerve terminals using amperometric glutamate biosensor.
abstract: Glutamate is the major excitatory neurotransmitter in the central nervous system, which is involved in the main aspects of normal brain functioning. High-affinity Na(+)-dependent glutamate transporters is key proteins, which transport extracellular glutamate to the cytoplasm of nerve cells, thereby preventing continuous activation of glutamate receptors, and thus the development of neurotoxicity. Disturbance in glutamate uptake is involved in the pathogenesis of major neurological disorders. Amperometric biosensors are the most promising and successful among electrochemical biosensors. In this study, we developed (1) amperometric glutamate biosensor, (2) methodological approach for the analysis of glutamate uptake in liquid samples of isolated rat brain nerve terminals (synaptosomes). The basal level of glutamate, the initial velocity of glutamate uptake and time-dependent accumulation of glutamate by synaptosomes were determined using developed glutamate biosensor. Comparative analysis of the data with those obtained by radioactive analysis, spectrofluorimetry and ion exchange chromatography was performed. Therefore, the methodological approach for monitoring of the velocity of glutamate uptake, which takes into consideration the definite level of endogenous glutamate in nerve terminals, was developed using glutamate biosensor.
authors: Laboratory of Biomolecular Electronics, Department of Translation Mechanisms of Genetic Information, Institute of Molecular Biology and Genetics, NAS of Ukraine, 150 Zabolotnogo Str., Kyiv 03680, Ukraine. Electronic address: alex_sold@yahoo.com.
authors: O
authors: O
authors: Soldatkin
authors: The Department of Neurochemistry, Palladin Institute of Biochemistry, NAS of Ukraine, 9 Leontovicha Street, Kyiv 01601, Ukraine.
authors: A
authors: A
authors: Nazarova
authors: The Department of Neurochemistry, Palladin Institute of Biochemistry, NAS of Ukraine, 9 Leontovicha Street, Kyiv 01601, Ukraine.
authors: N
authors: N
authors: Krisanova
authors: The Department of Neurochemistry, Palladin Institute of Biochemistry, NAS of Ukraine, 9 Leontovicha Street, Kyiv 01601, Ukraine.
authors: A
authors: A
authors: Borуsov
authors: Institute of High Technologies, Taras Shevchenko National University of Kyiv, 64, Volodymyrska Str., Kyiv 01003, Ukraine.
authors: D
authors: D
authors: Kucherenko
authors: Laboratory of Biomolecular Electronics, Department of Translation Mechanisms of Genetic Information, Institute of Molecular Biology and Genetics, NAS of Ukraine, 150 Zabolotnogo Str., Kyiv 03680, Ukraine.
authors: I
authors: I
authors: Kucherenko
authors: The Department of Neurochemistry, Palladin Institute of Biochemistry, NAS of Ukraine, 9 Leontovicha Street, Kyiv 01601, Ukraine.
authors: N
authors: N
authors: Pozdnyakova
authors: Laboratory of Biomolecular Electronics, Department of Translation Mechanisms of Genetic Information, Institute of Molecular Biology and Genetics, NAS of Ukraine, 150 Zabolotnogo Str., Kyiv 03680, Ukraine; Institute of High Technologies, Taras Shevchenko National University of Kyiv, 64, Volodymyrska Str., Kyiv 01003, Ukraine.
authors: A
authors: A
authors: Soldatkin
authors: The Department of Neurochemistry, Palladin Institute of Biochemistry, NAS of Ukraine, 9 Leontovicha Street, Kyiv 01601, Ukraine.
authors: T
authors: T
authors: Borisova
mesh_terms: D000818:Animals; D015374:Biosensing Techniques; D001921:Brain / Q000166:cytology; D004566:Electrodes; D005969:Glutamate Dehydrogenase / Q000378:metabolism; D018698:Glutamic Acid / Q000032:analysis* / Q000378:metabolism; D008297:Male; D009243:NAD / Q000378:metabolism; D010088:Oxidoreductases; D010984:Platinum; D051381:Rats; D017208:Rats, Wistar; D013574:Synaptosomes / Q000378:metabolism*
chemical_list: D009243:NAD; D018698:Glutamic Acid; D010984:Platinum; D010088:Oxidoreductases; D005969:Glutamate Dehydrogenase
keywords: Amperometry; Biosensor; Glutamate; Nerve terminals; Synaptosomes