Insects have been used extensively used as a model organism in genetics research and has significantly contributed to understanding molecular, cellular and evolutionary aspects of human behavior. However, sampling and chemical measurements of these mass-limited samples are still challenging. Research has focused on developing methods to obtain highly sensitive chemical quantification along with spatiotemporal information. The goal of chemical analysis is to relate neurochemical changes to either genetic mutations or to treatments that alter the state of the organism. Here, we develop methodology to rapidly quantify neurotransmitter and metabolite molecules. Chemical derivitization of biological molecules coupled with mass spectrometric detection will be used to measure neurotransmitter concentrations from individual cultured neurons. The methods presented are broadly applicable and can be used with a wide range of analytes. Additionally, this scheme simplifies detection and makes rapid relative quantification possible, allowing comprehensive and rapid analysis of neurotransmitters. This enables the analysis of attomole amounts of neurotransmitters and metabolites. In whole animals, fast changes in the extracellular concentration of neurotransmitters can arise from phasic neuronal firing, whereas long-lasting changes are associated with tonic firing.  We introduce a technique; to make measurements of the absolute concentration of electroactive neurotransmitters in vivo with 10-second time resolution.  This technique is used to measure slow (tonic) changes in real-time and coupled with traditional fast-scan cyclic voltammetry to measure rapid (phasic) changes in neurotransmission.  The pharmacology measurements validate the method and allow phasic and tonic concentrations to be tracked simultaneously at the same site in vivo.

Quantitative analysis of neurotransmitters in primary neuronal cultures and behaving animals

248th National Meeting (2014)

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