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 Citationer (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.

Originalsprog	Engelsk
Tidsskrift	Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
Vol/bind	1028
Sider (fra-til)	222-230
Antal sider	9
ISSN	1570-0232
DOI	https://doi.org/10.1016/j.jchromb.2016.05.048
Status	Udgivet - 15 aug. 2016

Emneord

Det Sundhedsvidenskabelige Fakultet
Monoamines
HPLC
Electrochemical detection
Cell culture
Guinea pig

Adgang til dokumentet

10.1016/j.jchromb.2016.05.048

Citationsformater

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.

I: Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences, Bind 1028, 15.08.2016, s. 222-230.

Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › 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, bind 1028, s. 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. I: Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences. 2016 ; Bind 1028. s. 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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