title: The potential of metabolomic analysis techniques for the characterisation of α1-adrenergic receptors in cultured N1E-115 mouse neuroblastoma cells.
abstract: Several studies of neuropathic pain have linked abnormal adrenergic signalling to the development and maintenance of pain, although the mechanisms underlying this are not yet fully understood. Metabolomic analysis is a technique that can be used to give a snapshot of biochemical status, and can aid in the identification of the mechanisms behind pathological changes identified in cells, tissues and biological fluids. This study aimed to use gas chromatography-mass spectrometry-based metabolomic profiling in combination with reverse transcriptase-polymerase chain reaction and immunocytochemistry to identify functional α1-adrenergic receptors on cultured N1E-115 mouse neuroblastoma cells. The study was able to confirm the presence of mRNA for the α1D subtype, as well as protein expression of the α1-adrenergic receptor. Furthermore, metabolomic data revealed changes to the metabolite profile of cells when exposed to adrenergic pharmacological intervention. Agonist treatment with phenylephrine hydrochloride (10 µM) resulted in altered levels of several metabolites including myo-inositol, glucose, fructose, alanine, leucine, phenylalanine, valine, and n-acetylglutamic acid. Many of the changes observed in N1E-115 cells by agonist treatment were modulated by additional antagonist treatment (prazosin hydrochloride, 100 µM). A number of these changes reflected what is known about the biochemistry of α1-adrenergic receptor activation. This preliminary study therefore demonstrates the potential of metabolomic profiling to confirm the presence of functional receptors on cultured cells.
authors.affiliation: School of Veterinary and Life Sciences, Murdoch University, Perth, WA, Australia.
authors.forename: Maria I
authors.initials: MI
authors.lastname: Wenner
authors.affiliation: School of Veterinary and Life Sciences, Murdoch University, Perth, WA, Australia. G.Maker@murdoch.edu.au.|Metabolomics Australia, Murdoch University, Perth, WA, Australia. G.Maker@murdoch.edu.au.|Separation Science and Metabolomics Laboratory, Murdoch University, Perth, WA, Australia. G.Maker@murdoch.edu.au.
authors.forename: Garth L
authors.initials: GL
authors.lastname: Maker
authors.affiliation: School of Veterinary and Life Sciences, Murdoch University, Perth, WA, Australia.|School of Psychology and Exercise Science, Murdoch University, Perth, WA, Australia.
authors.forename: Linda F
authors.initials: LF
authors.lastname: Dawson
authors.affiliation: School of Psychology and Exercise Science, Murdoch University, Perth, WA, Australia.
authors.forename: Peter D
authors.initials: PD
authors.lastname: Drummond
authors.affiliation: School of Veterinary and Life Sciences, Murdoch University, Perth, WA, Australia.
authors.forename: Ian
authors.initials: I
authors.lastname: Mullaney
keywords: Functional receptor; Metabolomics; N1E-115; Neuroblastoma; α1-Adrenergic receptor