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Quantitation of neurotransmitters and metabolites in microdialysate fluid and brain using advanced gas chromatographymass spectrometry (GC-MS)

Conference Paper


Abstract


  • Purpose: Sensitive and reliable measurement of neurotransmitters and

    other metabolites in brain is important for examining changes in brain

    biochemistry and metabolism. Our aim was to develop reliable analytical

    protocols for accurate measurement of different neurotransmitters and

    metabolites in cerebral spinal fluid (CSF) collected via microdialysis,

    focusing initially on several amino acids relevant for neurotransmission.

    The effect of potassium stimulation as a physiological control on

    extracellular levels of these compounds was also investigated. Method:

    CSF samples from the hippocampus and prefrontal cortex of male

    C57BL/6 mice (n=6 per region) were collected via microdialysis (flow

    rate 1 μl/min), before and after an infusion of artificial CSF containing

    50 nM K+. Following heavy isotopic dilution, metabolite extraction and

    derivatisation, samples were analysed using a triple quadrupole GC-MS

    and quantitated using calibration standards. Results: 10 μl CSF dialysate

    was sufficient for accurate measurement of several neurotransmitters and

    several metabolites including GABA (mean 40 nM ±6 SEM), glutamate

    (45 nM ±4), alanine, dopamine, glycine and serine. Stimulation with 50 nM

    K+ induced a significant 2-3-fold increase of alanine, GABA and glycine

    (p<0.05). Conclusions: GC-MS targeted analysis provides a sensitive

    tool for measuring a large range of neurotransmitters and metabolites

    during a single analysis in small aliquots of CSF. Small amounts of

    brain, plasma and other tissues can also be analysed. We are currently

    monitoring metabolic changes in a transgenic R6/1 mouse model of

    Huntingtons disease to examine mechanisms of neurodegeneration and

    identify potential biomarkers.

Authors


  •   Engel, Martin
  •   Frank, Elisabeth T. (external author)
  •   Jenner, Andrew M. (external author)

Publication Date


  • 2013

Citation


  • Engel, M., Frank, E. & Jenner, A. M. (2013). Quantitation of neurotransmitters and metabolites in microdialysate fluid and brain using advanced gas chromatographymass spectrometry (GC-MS). 33rd Meeting of the Australian Neuroscience Society: Program, Abstracts & List of Registrants (pp. 128-128). Australia: ANS.

Ro Metadata Url


  • http://ro.uow.edu.au/smhpapers/906

Start Page


  • 128

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  • 128

Abstract


  • Purpose: Sensitive and reliable measurement of neurotransmitters and

    other metabolites in brain is important for examining changes in brain

    biochemistry and metabolism. Our aim was to develop reliable analytical

    protocols for accurate measurement of different neurotransmitters and

    metabolites in cerebral spinal fluid (CSF) collected via microdialysis,

    focusing initially on several amino acids relevant for neurotransmission.

    The effect of potassium stimulation as a physiological control on

    extracellular levels of these compounds was also investigated. Method:

    CSF samples from the hippocampus and prefrontal cortex of male

    C57BL/6 mice (n=6 per region) were collected via microdialysis (flow

    rate 1 μl/min), before and after an infusion of artificial CSF containing

    50 nM K+. Following heavy isotopic dilution, metabolite extraction and

    derivatisation, samples were analysed using a triple quadrupole GC-MS

    and quantitated using calibration standards. Results: 10 μl CSF dialysate

    was sufficient for accurate measurement of several neurotransmitters and

    several metabolites including GABA (mean 40 nM ±6 SEM), glutamate

    (45 nM ±4), alanine, dopamine, glycine and serine. Stimulation with 50 nM

    K+ induced a significant 2-3-fold increase of alanine, GABA and glycine

    (p<0.05). Conclusions: GC-MS targeted analysis provides a sensitive

    tool for measuring a large range of neurotransmitters and metabolites

    during a single analysis in small aliquots of CSF. Small amounts of

    brain, plasma and other tissues can also be analysed. We are currently

    monitoring metabolic changes in a transgenic R6/1 mouse model of

    Huntingtons disease to examine mechanisms of neurodegeneration and

    identify potential biomarkers.

Authors


  •   Engel, Martin
  •   Frank, Elisabeth T. (external author)
  •   Jenner, Andrew M. (external author)

Publication Date


  • 2013

Citation


  • Engel, M., Frank, E. & Jenner, A. M. (2013). Quantitation of neurotransmitters and metabolites in microdialysate fluid and brain using advanced gas chromatographymass spectrometry (GC-MS). 33rd Meeting of the Australian Neuroscience Society: Program, Abstracts & List of Registrants (pp. 128-128). Australia: ANS.

Ro Metadata Url


  • http://ro.uow.edu.au/smhpapers/906

Start Page


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