FLUORESCENCE MICROSCOPY ASSAY FOR REAL-TIME MONITORING OF NEURO-PHARMACOLOGICAL MODULATION OF TRANSMEMBRANE SIGNAL TRANSDUCTION PATHWAYS

ONE OF THE MAJOR DIFFICULTIES IN DETERMINING THE ACTION AND SPECIFICITY OF PHARMACEUTICAL AGENTS IS THE LIMITATIONS OF AVAILABLE BIOLOGICAL ANALYTICAL SYSTEMS. WE ARE DEVELOPING A FLUORESCENCE MICROSCOPY ASSAY SYSTEM WHICH WILL BE CAPABLEOF MONITORING CHANGES IN IMPORTANT PHYSIOLOGICAL TRANSDUCTION PROCESSES IN SINGLE CELLS. DURING PHASE I, A FLUORESCENT PHOSPHOLIPID SUBSTRATE FOR THE ENZYME, PHOSPHOLIPASE A2, IS TO BE INCORPORATED INTO CULTURED CELLS TO DETERMINE IF, UPON ITS DRUG-RELATED METABOLISM, THERE IS A CHANGE IN THE FLUORESCENT PROBES PROPERTIES. ULTIMATELY, THIS ASSAY IS TO BE APPLIED TO MORE COMPLEX NEURAL SYSTEMS (E.G. BRAIN HIPPOCAMPAL SLICES) TO PROVIDE REAL-TIME INFORMATION BOTH ON RECEPTOR SPECIFICITY AND CONSIDERABLE DETAIL ON THE NETWORKS INVOLVED AND AFFECTED IN THE TRANSMISSION PROCESS. THIS TYPE OF INFORMATION IS NOT READILY AVAILABLE FROM EXISTING ANALYTICAL TESTS. IMPLEMENTATION OF THIS ASSAY DEPENDS ON THE DEVELOPMENT OF AHIGH RESOLUTION MICROSCOPY SYSTEM WITH MORE EFFICIENT LIGHT TRANSMISSION THAN EXISTING COMMERCIAL MICROSCOPES. ALSO, DEVELOPMENT OF THIS SYSTEM WILL ALLOW US TO PROVIDE THIS SYSTEM AS A ROUTINE SERVICE OR TOOL FOR ANALYSES OF MECHANISM OF ACTION OF A MULTITUDE OF PSYCHOACTIVE DRUGS. ONE OF THE MAJOR DIFFICULTIES IN DETERMINING THE ACTION AND SPECIFICITY OF PHARMACEUTICAL AGENTS IS THE LIMITATIONS OF AVAILABLE BIOLOGICAL ANALYTICAL SYSTEMS. WE ARE DEVELOPING A FLUORESCENCE MICROSCOPY ASSAY SYSTEM WHICH WILL BE CAPABLEOF MONITORING CHANGES IN IMPORTANT PHYSIOLOGICAL TRANSDUCTION PROCESSES IN SINGLE CELLS. DURING PHASE I, A FLUORESCENT PHOSPHOLIPID SUBSTRATE FOR THE ENZYME, PHOSPHOLIPASE A2, IS TO BE INCORPORATED INTO CULTURED CELLS TO DETERMINE IF, UPON ITS DRUG-RELATED METABOLISM, THERE IS A CHANGE IN THE FLUORESCENT PROBES PROPERTIES. ULTIMATELY, THIS ASSAY IS TO BE APPLIED TO MORE COMPLEX NEURAL SYSTEMS (E. G. BRAIN HIPPOCAMPAL SLICES) TO PROVIDE REAL-TIME INFORMATION BOTH ON RECEPTOR SPECIFICITY AND CONSIDERABLE DETAIL ON THE NETWORKS INVOLVED AND AFFECTED IN THE TRANSMISSION PROCESS. THIS TYPE OF INFORMATION IS NOT READILY AVAILABLE FROM EXISTING ANALYTICAL TESTS. IMPLEMENTATION OF THIS ASSAY DEPENDS ON THE DEVELOPMENT OF AHIGH RESOLUTION MICROSCOPY SYSTEM WITH MORE EFFICIENT LIGHT TRANSMISSION THAN EXISTING COMMERCIAL MICROSCOPES. ALSO, DEVELOPMENT OF THIS SYSTEM WILL ALLOW US TO PROVIDE THIS SYSTEM AS A ROUTINE SERVICE OR TOOL FOR ANALYSES OF MECHANISM OF ACTION OF A MULTITUDE OF PSYCHOACTIVE DRUGS.