Simultaneous quantification of monoamine neurotransmitters and their biogenic metabolites intracellularly and extracellularly in primary neuronal cell cultures and in sub-regions of guinea pig brain

Anne Marie Voigt Schou-Pedersen; Stine Normann Hansen; Pernille Tveden-Nyborg; Jens Lykkesfeldt

doi:10.1016/j.jchromb.2016.05.048

Simultaneous quantification of monoamine neurotransmitters and their biogenic metabolites intracellularly and extracellularly in primary neuronal cell cultures and in sub-regions of guinea pig brain

Anne Marie Voigt Schou-Pedersen, Stine Normann Hansen, Pernille Tveden-Nyborg, Jens Lykkesfeldt

Lifepharm

9 Citations (Scopus)

Abstract

In the present paper, we describe a validated chromatographic method for the simultaneous quantification of monoamine neurotransmitters and their biogenic metabolites intracellularly and extracellularly in primary neuronal cell culture and in sub-regions of the guinea pig brain. Electrochemical detection provided limits of quantifications (LOQs) between 3.6 and 12 nM. Within the linear range, obtained recoveries were from 90.9 ± 9.9 to 120 ± 14% and intra-day and inter-day precisions found to be less than 5.5% and 12%, respectively. The analytical method was applicable for quantification of intracellular and extracellular amounts of monoamine neurotransmitters and their metabolites in guinea pig frontal cortex and hippocampal primary neuronal cell cultures. Noradrenaline, dopamine and serotonin were found to be in a range from 0.31 to 1.7 pmol per 2 million cells intracellularly, but only the biogenic metabolites could be detected extracellularly. Distinct differences in monoamine concentrations were observed when comparing concentrations in guinea pig frontal cortex and cerebellum tissue with higher amounts of dopamine and its metabolites, 3,4-dihydroxyphenylacetic acid and homovanillic acid in frontal cortex, as compared to cerebellum. The chemical turnover in frontal cortex tissue of guinea pig was for serotonin successfully predicted from the turnover observed in the frontal cortex cell culture. In conclusion, the present analytical method shows high precision, accuracy and sensitivity and is broadly applicable to monoamine measurements in cell cultures as well as brain biopsies from animal models used in preclinical neurochemistry.

Original language	English
Journal	Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
Volume	1028
Pages (from-to)	222-230
Number of pages	9
ISSN	1570-0232
DOIs	https://doi.org/10.1016/j.jchromb.2016.05.048
Publication status	Published - 15 Aug 2016

Keywords

Faculty of Health and Medical Sciences

Access to Document

10.1016/j.jchromb.2016.05.048

Cite this

Schou-Pedersen, A. M. V., Hansen, S. N., Tveden-Nyborg, P., & Lykkesfeldt, J. (2016). Simultaneous quantification of monoamine neurotransmitters and their biogenic metabolites intracellularly and extracellularly in primary neuronal cell cultures and in sub-regions of guinea pig brain. Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences, 1028, 222-230. https://doi.org/10.1016/j.jchromb.2016.05.048

Simultaneous quantification of monoamine neurotransmitters and their biogenic metabolites intracellularly and extracellularly in primary neuronal cell cultures and in sub-regions of guinea pig brain. / Schou-Pedersen, Anne Marie Voigt; Hansen, Stine Normann; Tveden-Nyborg, Pernille et al.
In: Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences, Vol. 1028, 15.08.2016, p. 222-230.

Research output: Contribution to journal › Journal article › Research › peer-review

Schou-Pedersen, AMV, Hansen, SN, Tveden-Nyborg, P & Lykkesfeldt, J 2016, 'Simultaneous quantification of monoamine neurotransmitters and their biogenic metabolites intracellularly and extracellularly in primary neuronal cell cultures and in sub-regions of guinea pig brain', Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences, vol. 1028, pp. 222-230. https://doi.org/10.1016/j.jchromb.2016.05.048

Schou-Pedersen AMV, Hansen SN, Tveden-Nyborg P , Lykkesfeldt J. Simultaneous quantification of monoamine neurotransmitters and their biogenic metabolites intracellularly and extracellularly in primary neuronal cell cultures and in sub-regions of guinea pig brain. Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences. 2016 Aug 15;1028:222-230. doi: 10.1016/j.jchromb.2016.05.048

Schou-Pedersen, Anne Marie Voigt ; Hansen, Stine Normann ; Tveden-Nyborg, Pernille et al. / Simultaneous quantification of monoamine neurotransmitters and their biogenic metabolites intracellularly and extracellularly in primary neuronal cell cultures and in sub-regions of guinea pig brain. In: Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences. 2016 ; Vol. 1028. pp. 222-230.

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abstract = "In the present paper, we describe a validated chromatographic method for the simultaneous quantification of monoamine neurotransmitters and their biogenic metabolites intracellularly and extracellularly in primary neuronal cell culture and in sub-regions of the guinea pig brain. Electrochemical detection provided limits of quantifications (LOQs) between 3.6 and 12 nM. Within the linear range, obtained recoveries were from 90.9 ± 9.9 to 120 ± 14% and intra-day and inter-day precisions found to be less than 5.5% and 12%, respectively. The analytical method was applicable for quantification of intracellular and extracellular amounts of monoamine neurotransmitters and their metabolites in guinea pig frontal cortex and hippocampal primary neuronal cell cultures. Noradrenaline, dopamine and serotonin were found to be in a range from 0.31 to 1.7 pmol per 2 million cells intracellularly, but only the biogenic metabolites could be detected extracellularly. Distinct differences in monoamine concentrations were observed when comparing concentrations in guinea pig frontal cortex and cerebellum tissue with higher amounts of dopamine and its metabolites, 3,4-dihydroxyphenylacetic acid and homovanillic acid in frontal cortex, as compared to cerebellum. The chemical turnover in frontal cortex tissue of guinea pig was for serotonin successfully predicted from the turnover observed in the frontal cortex cell culture. In conclusion, the present analytical method shows high precision, accuracy and sensitivity and is broadly applicable to monoamine measurements in cell cultures as well as brain biopsies from animal models used in preclinical neurochemistry.",

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