Date of Award

12-2023

Document Type

Open Access Thesis

Department

Biological Science

First Advisor

Theresa Curtis

Abstract

Non-olfactory cells have excellent biosensor potential because they express functional olfactory receptors (ORs) and are non-neuronal cells that are easy to culture. ORs are G-protein coupled receptors (GPCRs), and there is a well-established link between different classes of G-proteins and cytoskeletal structure changes affecting cellular morphology that has been unexplored for odorant sensing. Thus, the present study was conducted to determine if odorant binding in non-olfactory cells causes cytoskeletal changes that will lead to cell changes detectable by electric cell-substrate impedance sensing (ECIS). To this end, we used the human umbilical vein endothelial cells (HUVECs), which express OR10J5, and the human keratinocyte (HaCaT) cells, which express OR2AT4. Using these two different cell barriers, we showed that odorant addition, lyral and Sandalore, respectively, caused an increase in cAMP, changes in the organization of the cytoskeleton, and a decrease in the integrity of the junctions between the cells, causing a decrease in cellular electrical resistance. In addition, the random cellular movement of the monolayers (micromotion) was significantly decreased after odorant exposure. Collectively, these data demonstrate a new physiological role of olfactory receptor signaling in endothelial and epithelial cell barriers and represent a new label-free method to detect odorant binding.

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