Chromatographic assay to study the activity of multiple enzymes involved in the synthesis and metabolism of dopamine and serotonin

Morgan, Lindsay D., Baker, Hannah, Yeoman, Mark S. and Patel, Bhavik Anil (2012) Chromatographic assay to study the activity of multiple enzymes involved in the synthesis and metabolism of dopamine and serotonin. Analyst, 137 (6). pp. 1409-1415. ISSN 0003-2654

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Abstract

Serotonin and dopamine are crucial regulators of signalling in the peripheral and central nervous systems. We present an ex-vivo, isocratic chromatographic method that allows for the measurement of tyrosine, L-3,4-dihydroxyphenylalanine (L-DOPA), dopamine, 3,4-dihydroxyphenylacetic acid (DOPAC), tryptophan, 5-hydroxytryptophan (5-HTP), serotonin and 5-hydroxy-3-indoleacetic acid (5-HIAA) in a model central nervous (CNS) system, to study the role of key enzymes involved in the synthesis and metabolism of serotonin and dopamine. By utilising a sample splitting technique, we could test a single CNS sample at multiple time points under various pharmacological treatments. In, addition, we were able to conduct this assay by utilising the endogenous biochemical components of the CNS to study the synthesis and metabolism of serotonin and dopamine, negating the requirement of additional enzyme activators or stabilisers in the biological matrix. Finally we utilised NSD-1015, an aromatic amino acid decarboxylase enzyme inhibitor used to study the synthesis of dopamine and serotonin to monitor alterations in levels of key neurochemicals. 3-hydroxybenzylhydrazine dihydrochloride (NSD-1015) was able to reduce levels of serotonin and dopamine, whilst elevating precursors L-DOPA and 5-HTP.

Item Type:	Article
Additional Information:	Funding information: BAP and MSY would like acknowledge support from BBRSC grant (BB/G015147/1).
Faculty \ School:	Faculty of Science > School of Chemistry, Pharmacy and Pharmacology
Depositing User:	LivePure Connector
Date Deposited:	06 Dec 2024 01:43
Last Modified:	06 Dec 2024 01:43
URI:	https://ueaeprints.uea.ac.uk/id/eprint/97929
DOI:	10.1039/C2AN16227J

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