Enhancement, modulation and electrophysiological characterization of murine olfactory neurons to odorant stimulation in vitro

The objectives of this study are to electrophysiologically characterize cultured olfactory epithelium (OE) and to compare with that of rapidly isolated OE; to establish the Galphai involvement in the modulation of adenylyl cyclase (AC) and to demonstrate the participation of zinc in odorant stimulated enhancement of smell. The murine family has a well developed sense of smell and is widely used in the field of olfactory research. Producing a long lived, readily accessible, economically feasible and physiologically viable cultured OE model would be ideal, to meet with specific requirements and nature of the research.; The cultured OE of different ages were exposed to charcoal-filtered air, individual odorants, and an odorant mixture of (+)carvone, (-)carvone, eugenol, and ethyl butyrate at varying concentrations or with varying inter pulse periods to explore the system's ability to respond to and adapt to odorant presentation.; The study also involved with Galphai-Protein Inhibition of olfactory signal transduction. The whole-cell voltage clamp recordings of odorant-stimulated olfactory neurons indicate that endogenous Gi protein negatively regulates odorant-evoked intracellular signaling. Rat OE were subjected to whole cell recordings with holding membrane potential of +/- 57 Mv. The patch electrode (resistance of 8+/-3MO) was filled with intracellular fluid (ICF) in the presence or absence of antibodies directed against the Galphas/olf-subunits (Santa Cruz Biotech.), antibodies raised against Galphai-subunits (Calbiochem) or control antibodies.; In addition to G proteins involvement in the enhancement, modulation and inhibition of Odorant signals, there is evidence that metals like zinc enhances odorant induced response of Rat OE are exposed to crystalline metallic nanoparticles which are found in proteon nucleating centers (PNC) and metallic Zinc nanoparticles (artificial PNC). EOG recordings were made from dissected rat olfactory receptor neurons under control conditions (in the presence of extra cellular fluid---ECF) and experimental conditions (in the presence of PNC and Zn and Cu). The metal nanoparticles were integrated with the odorants of interest and were delivered onto the odor receptors.; Our results show that in cultured tissues the characteristics of the EOG responses are similar to that of standard EOG responses of dissected OE. Cultures one week old or less (n = 8) showed rapid rise times but prolonged decay times. Cultures aged 13 to 15 days in vitro (n = 8) showed both rapid rise and decay times. (Abstract shortened by UMI.)...