This subproject is one of many research subprojects utilizing theresources provided by a Center grant funded by NIH/NCRR. The subproject andinvestigator (PI) may have received primary funding from another NIH source,and thus could be represented in other CRISP entries. The institution listed isfor the Center, which is not necessarily the institution for the investigator.The fruit fly Drosophila melanogaster has been utilized as a model in biology due to its amenability to genetic manipulation. Fruit flies can form memories after appropriate associative olfactory learning protocols. These processes can be dissected into different temporal phases using mutants with specific spatial expression in central brain structures. With the ability to perform genetic manipulation and established behavioral protocols, examining the physiology of the brain areas involved is the next step in the development of this important model. The Mushroom Bodies, (MB's), are paired groups of neurons on the posterior dorsal surface of the fly brain. These receive chemosensory information from the antennal lobes. Any disruption of the MB's causes significant effects to associative olfactory learning and memory. To complete this model system I developed a preparation to allow the monitoring of MB function during application of physiological stimuli, odors. However, this did not allow direct pharmacological analysis due to technical limitations. The BRIN/INBRE programs have allowed me to equip my laboratory with an imaging system and develop a new variation of this technique that does allow direct manipulation. We have now been able to record quantitave responses to iontophoretically applied acetylcholine, (ACh), using the endogenous fluorescent calcium reporter camgaroo, expressed exclusively in MB neurons, (ACh is thought to be the transmitter mediating the odor response in the MBs from the antennal lobe projections). Increases in calcium can be reversibly blocked by tubocurare and eliminated in zero calcium, (Wright, 2006). The synaptic physiology of these neurons, and the underlying signal transduction mechanisms, can now be investigated for the first time in a living fly. Currently, we are co-applying ACh and dopamine, (DA), to investigate the mechanisms underlying associative learning; DA is thought to mediate the conditioning, noxious stimulus. This will add to our knowledge of chemosensory perception and learning and memory.
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