[Home ] [Archive]   [ فارسی ]  
:: Main :: About :: Current Issue :: Archive :: Search :: Submit :: Contact ::
Main Menu
Home::
Journal Information::
Articles archive::
Publication Ethics::
Peer Review Process::
Indexing Databases::
For Authors::
For Reviewers::
Subscription::
Contact us::
Site Facilities::
::
Google Scholar Metrics

Citation Indices from GS

AllSince 2019
Citations64463744
h-index2720
i10-index18983

..
Search in website

Advanced Search
..
Receive site information
Enter your Email in the following box to receive the site news and information.
..
Registered in

AWT IMAGE

AWT IMAGE

..
:: Volume 31, Issue 2 (6-2023) ::
Journal of Ilam University of Medical Sciences 2023, 31(2): 65-75 Back to browse issues page
Measurement of Glutamate Neurotransmitter in the Brain of Male Rat Using Glutamate Oxidase-Based Electrochemical Biosensor
Faezeh Faraji1 , Hassan Tavakoli * 2, Mahvash Jafari3 , Akram Eidi1 , Adeleh Divsalar4
1- Dept of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran
2- Radiation Injuries Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran , Tavakoli@bmsu.ac.ir
3- Dept of Biochemistry, Faculty of Medicine, Baqiyatallah University of Medical Sciences, Tehran, Iran
4- Dept of Cell and Molecular Sciences, Faculty of Biological Sciences, Kharazmi University, Tehran, Iran
Abstract:   (966 Views)
Introduction: Glutamate oxidase (GluOx; EC 1.4.3.11), as an oxidoreductase enzyme, catalyzes the oxidation of glutamate to α-ketoglutarate, ammonia, and hydrogen peroxide (H2O2). In this reaction, the amount of H2O2 is proportional to the concentration of glutamate and its concentration can be measured by using an electrochemical biosensor. The same as other enzyme-based biosensors, glutamate oxidase is one of the key elements in the construction of glutamate biosensors. Such biosensors are fully capable of identifying biological analytes, such as glutamine, ammonia, and creatinine. In addition, glutamate oxidase-based biosensors have many applications in quantitative and qualitative measurements in analytical chemistry, determining the quality of food products, as well as early detection of heart and liver disorders in clinical biochemistry. Considering the importance of the glutamate neurotransmitter in various brain functions, this study investigated its measurement in the brain of male Wistar rats by using a glutamate oxidase-based biosensor.
Material & Methods: In order to measure glutamate, initially, a glutamate oxidase-based biosensor was made by simultaneously immobilizing the enzyme and chitosan on the platinum electrode surface. Then, the animals were sacrificed, and their brains were removed and placed in a phosphate buffer. Afterward, the contents of the brain were centrifuged to create a completely uniform mixture. Finally, the concentration of glutamate in the prepared brain samples was measured using the fabricated biosensor by the cyclic voltammetry technique.
Findings: The results of cyclic voltammetry experiments showed that the cathodic peak current of the fabricated biosensor was 0.812 µA. Moreover, the calibration curve indicated that the biosensor response was linear up to 1 mM and glutamate concentration in brain samples was also equal to 63.5 µM.
Discussion & Conclusion: The cyclic voltammetry experiments showed that the concentration of H2O2, which is produced during the catalytic activity of glutamate oxidase, is completely proportional to the concentration of glutamate oxidase. Furthermore, this study showed that, despite the very low concentration of glutamate neurotransmitters, the glutamate oxidase-based electrochemical biosensor can precisely identify it qualitatively and quantitatively in biological samples, such as brain samples.
Keywords: Chitosan, Cyclic voltammetry, Electrochemical biosensor, Glutamate oxidase, Glutamate
Full-Text [PDF 1115 kb]   (425 Downloads)    
Type of Study: Research | Subject: clinical biochemistry
Received: 2022/12/4 | Accepted: 2023/03/5 | Published: 2023/06/5
References
1. Gasparini C, Smith R, Griffiths L. Biochemical studies of the neurotransmitter glutamate: A key player in migraine. Austin J Clin Neurol 2015; 2: 1079 1-8.
2. Nguyen TN, Nolan JK, Park H, Lam s, Fattah M, Page JC, et al. Facile fabrication of flexible glutamate biosensor using direct writing of platinum nanoparticle-based nanocomposite ink. Biosens Bioeletron X 2019; 131: 257-66. doi:10.1016/j.bios.2019.01.051.
3. Meng L,Wu G, Chen G, Cai C, Sun Y , Yuan Z. Low potential detection of glutamate based on the electrocatalytic oxidation of NADH at thionine/single-walled carbon nanotubes composite modified electrode. Biosens Bioelectron X 2009; 24: 1751-56. doi:10.1016/j.bios.2008.09.001.
4. Butterfield DA, Pocernich CB. The glutamatergic system and Alzheimer’s disease. CNS drugs 2003; 17: 641-52. doi:10.2165/00023210-200317090-00004.
5. Lewerenz J, Maher P. Chronic glutamate toxicity in neurodegenerative diseases-what is the evidence? Front Neurosci 2015; 9: 469. doi:10.3389/fnins.2015.00469.
6. Paulose C, Amee K, Anu J. Neurotransmitters functional balance in neurodegenerative disease management: Recent Advances. Sci Soc 2006; 5: 23-30.
7. Lau A, Tymianski M. Glutamate receptors, neurotoxicity and neurodegeneration. Pflugers Arch 2010; 460: 525-42. doi:10.1007/s00424-010-0809-1.
8. Black DW, Andreasen NC. Introductory textbook of psychiatry. American Psychiatric Pub 2011.
9. Batra B, Yadav M, Pundir CS. l-Glutamate biosensor based on l-glutamate oxidase immobilized onto ZnO nanorods/polypyrrole modified pencil graphite electrode. Biochem Eng J 2016; 105: 428-36. doi:10.1016/j.bej.2015.10.012.
10. Dorozhko E, Korotkova EI, Shabaeva AA, Mosolkov AY. Electrochemical determination of L-glutamate on a carbon-containing electrode modified with gold by voltammetry. Procedia Chem 2015; 15: 365-70. doi:10.1016/j.proche.2015.10.058.
11. Kucherenko DY, Kucherenko IS, Soldatkin OO, Soldatkin AP. Application of glutamate-sensitive biosensor for analysis of foodstuff. Biotechnol Acta 2018; 11: 57-67.doi: 10.15407/biotech11.04.057.
12. Dalkıran B, Erden PE, Kılıç E. Graphene and tricobalt tetraoxide nanoparticles based biosensor for electrochemical glutamate sensing. Artif Cells Nanomed Biotechnol 2017; 45: 340-48. doi:10.3109/21691401.2016.1153482.
13. Monge-Acuña AA, Fornaguera-Trías J A. high performance liquid chromatography method with electrochemical detection of gamma-aminobutyric acid, glutamate and glutamine in rat brain homogenates. J Neurosci Methods 2009; 183: 176-81. doi:10.1016/j.jneumeth.2009.06.042.
14. Buck K, Voehringer P, Ferger B. Rapid analysis of GABA and glutamate in microdialysis samples using high performance liquid chromatography and tandem mass spectrometry. J Neurosci Methods 2009; 182: 78-84. doi:10.1016/j.jneumeth.2009.05.018.
15. Ganesana M, Trikantzopoulos E, Maniar Y, Lee ST, Venton BJ. Development of a novel micro biosensor for in vivo monitoring of glutamate release in the brain. Biosens Bioelectron X 2019; 130: 103-9. doi:10.1016/j.bios.2019.01.049.
16. Rocchitta G, Bacciu A, Arrigo P, Migheli R, Bazzu G, Serra PA. Propylene glycol stabilizes the linear response of glutamate biosensor: potential implications for in-vivo neurochemical monitoring. Chemosensors 2018; 6: 58. doi:10.20944/preprints201810.0630.v1.
17. Borisova T, Kucherenko D, Soldatkin O, Kucherenko I, Pastukhov A, Nazarova A, et al. An amperometric glutamate biosensor for monitoring glutamate release from brain nerve terminals and in blood plasma. Anal Chim Acta 2018; 1022: 113-23. doi:10.1016/j.aca.2018.03.015.
18. Soldatkina OV, Soldatkin OO, Kasap BO, Kucherenko DY, Kucherenko IS, Kurc BA, et al. A novel amperometric glutamate biosensor based on glutamate oxidase adsorbed on silicalite. Nanoscale Res Lett 2017; 12: 1-8. doi:10.1186/s11671-017-2026-8.
19. Zhang M, Mullens C, Gorski W. Amperometric glutamate biosensor based on chitosan enzyme film. Electrochim Acta 2006; 51: 4528-32. doi:10.1016/j.electacta.2006.01.010
20. Zhang M, Mullens C, Gorski W. Chitosan‐glutamate oxidase gels: synthesis, characterization, and glutamate determination. Electroanalysis 2005; 17: 2114-20.doi:10.1002/elan.200503348.
21. Tseng TTC, Chang CF, Chan WC. Fabrication of implantable, enzyme-immobilized glutamate sensors for the monitoring of glutamate concentration changes in vitro and in vivo. Molecules 2014; 19: 7341-55. [DOI:10.3390/molecules19067341.]
22. Kusakabe H, Midorikawa Y, Fujishima T, Kuninaka A, Yoshino H. Purification and properties of a new enzyme, l-glutamate oxidase, from Streptomyces sp. X-119-6 grown on wheat bran. Agric Biol Chem 1983; 47: 1323-28. doi:10.1080/00021369.1983.10866079.
23. Utsumi T, Arima J, Sakaguchi C, Tamura T, Sasaki C, Kusakabe H, et al. Arg305 of Streptomyces L-glutamate oxidase plays a crucial role for substrate recognition. Biochem Biophys Res Commun 2012; 417: 951-55. doi:10.1016/j.bbrc.2011.12.033.
24. Tao-Cheng JH, Gallant PE, Brightman MW, Dosemeci A, Reese TS. Structural changes at synapses after delayed perfusion fixation in different regions of the mouse brain. J Comp Neurol 2007; 501: 731-40. doi:10.1002/cne.21276.
25. Şimşek Ş, Aynacı E, Arslan F. An amperometric biosensor for L-glutamate determination prepared from L-glutamate oxidase immobilized in polypyrrole-polyvinylsulphonate film. Artif Cells Nanomed Biotechnol 2016; 44: 731-40. doi:10.1002/cne.21276.
Send email to the article author

Add your comments about this article
Your username or Email:

CAPTCHA

Ethics code: IR.BMSU.BAQ.REC.1401.015


XML   Persian Abstract   Print


Download citation:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:

Faraji F, Tavakoli H, Jafari M, Eidi A, Divsalar A. Measurement of Glutamate Neurotransmitter in the Brain of Male Rat Using Glutamate Oxidase-Based Electrochemical Biosensor. J. Ilam Uni. Med. Sci. 2023; 31 (2) :65-75
URL: http://sjimu.medilam.ac.ir/article-1-7804-en.html


Rights and permissions
Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
Volume 31, Issue 2 (6-2023) Back to browse issues page
مجله دانشگاه علوم پزشکی ایلام Journal of Ilam University of Medical Sciences
Persian site map - English site map - Created in 0.15 seconds with 41 queries by YEKTAWEB 4654