title: Chromatographic assay to study the activity of multiple enzymes involved in the synthesis and metabolism of dopamine and serotonin.
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.
authors.affiliation: Centre for Biomedical and Health Sciences Research, University of Brighton, School of Pharmacy and Biomolecular Sciences, Brighton, BN2 4GJ.
authors.forename: Lindsay D
authors.initials: LD
authors.lastname: Morgan
authors.forename: Hannah
authors.initials: H
authors.lastname: Baker
authors.forename: Mark S
authors.initials: MS
authors.lastname: Yeoman
authors.forename: Bhavik Anil
authors.initials: BA
authors.lastname: Patel
mesh_terms: D015102:3,4-Dihydroxyphenylacetic Acid / Q000737:chemistry / Q000378:metabolism; D006916:5-Hydroxytryptophan / Q000737:chemistry / Q000378:metabolism; D000818:Animals; D001681:Biological Assay / Q000379:methods*; D002490:Central Nervous System / Q000033:anatomy & histology / Q000737:chemistry / Q000378:metabolism; D002845:Chromatography / Q000379:methods*; D004298:Dopamine / Q000737:chemistry / Q000378:metabolism*; D015259:Dopamine Agents / Q000737:chemistry / Q000378:metabolism; D006897:Hydroxyindoleacetic Acid / Q000737:chemistry / Q000378:metabolism; D007980:Levodopa / Q000737:chemistry / Q000378:metabolism; D008195:Lymnaea / Q000033:anatomy & histology / Q000378:metabolism; D008954:Models, Biological; D015394:Molecular Structure; D012701:Serotonin / Q000737:chemistry / Q000378:metabolism*; D014364:Tryptophan / Q000737:chemistry / Q000378:metabolism; D014443:Tyrosine / Q000737:chemistry / Q000378:metabolism
chemical_list: D015259:Dopamine Agents; D015102:3,4-Dihydroxyphenylacetic Acid; D012701:Serotonin; D014443:Tyrosine; D007980:Levodopa; D006897:Hydroxyindoleacetic Acid; D014364:Tryptophan; D006916:5-Hydroxytryptophan; D004298:Dopamine