Development of an in vitro model system to study primary sensory transduction mechanisms.
AuthorStengl, Monika Anna-Helga.
AdvisorHildebrand, John G.
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PublisherThe University of Arizona.
RightsCopyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction or presentation (such as public display or performance) of protected items is prohibited except with permission of the author.
AbstractSex-pheromone components released by Manduca sexta females are detected solely by male-specific olfactory receptor neurons that innervate long sensilla trichodea on the male antennae. To facilitate studies of the development and physiology of these receptor cells, I have produced primary in vitro cultures of cells dissociated from pupal male antennae. These cultures comprise several morphological types of cells, two of these cell types could be characterized immunocytochemically with a pair of monoclonal antibodies that were shown previously to recognize certain antigens in olfactory receptor neurons at defined stages of development. The good correlation between in vivo and in vitro expression of these antigens suggest that the immunocytochemically recognized cells are olfactory receptor neurons that follow at least partially their normal course of development in vitro. Patch-clamp studies revealed that the immunocytochemically recognized olfactory receptor neurons express three different kinds of Cs⁺-blockable K⁺ channels and at least one kind of Tetrodotoxin-blockable Na⁺ channel after three weeks in vitro. At least one channel, an unspecific cation channel, responds with higher frequency openings after stimulation with female pheromone-gland extracts in vitro. Thus it could be demonstrated for the first time that identifiable cultured insect olfactory receptor neurons differentiate in morphological and physiological terms and are able to respond to pheromones in vitro, providing an ideal model system for studies of primary sensory transduction in vitro.
Degree ProgramMolecular and Cellular Biology