The long-term objective of this work is to develop enzyme microbiosensor-based instruments for rapid (<1 min) in vivo, intracellular detection of non-easily oxidized neurotransmitters. Specifically, in Phase II amperometric microbiosensors (< 5micro m O.D.) will be fabricated for detection of neurotransmitters glutamate, aspartate, gamma-aminobutyric acid (GABA), choline, acetylcholine, glycine and the amino acids glutamine, histidine, alanine, asparagine, valine, isoleucine, leucine, arginine, methionine, tryptophan and phenylalanine, which are possible neurotransmitters. These sensors will be used to study the rapid changes in analyte concentration that occur within neurons and in the extracellular fluid. Thus, for the first time, it will be possible to study the dynamics of processes such as neurotransmitter release and uptake within single neurons. The biosensor response in resting cells will be compared to that obtained by single cell LCEC analysis of the same cells. This technology will enable a vast increase in the fundamental knowledge of neuronal cellular processes, so aiding research into neurodegenerative diseases and facilitating the recovery of patients who have suffered brain damage.
