Author(s)
Adam Kimple, MD, PhD
Vince Setola, PhD
Staci Cohen, PhD
David Siderovsky, PhD
Affiliation(s)
University of North Carolina Chapel Hill
Abstract:
"Introduction: Mucociliary clearance (MCC) maintains the respiratory epithelium by propelling debris laden mucus over a ciliated epithelial monolayer for removal. Impairment of MCC promotes disease states such as chronic rhinosinusitis (CRS). Mutations in taste receptors have been shown to impair MCC and serve as an independent predictor of outcomes after sinus surgery. Standard assays for gustatory signaling are highly artificial, involving chimeric G-proteins heterologously expressed in immortalized kidney cells. Identification of cell lines that endogenously express gustatory signaling components will provide a more physiologic system to study gustation and allow for the development of high-throughput screens to identify modulators of gustation and MCC.
Methods: Twenty immortalized cell lines from the aerodigestive tract were obtained and cultured according to standard protocols. Ribonucleic acid was extracted and quantitative real-time polymerase chain reaction was used to detect transcription of taste receptors and downstream components. Selected cell lines, which demonstrated endogenous expression of the gustatory signaling cascade, were assayed using tastant induced changes in intracellular calcium and cAMP.
Results: Gustatory components are variably expressed in the immortalized aerodigestive cell lines. Application of tastants elicited a calcium and cAMP response in cell lines identified to express components of the gustatory cascade.
Conclusions: We have identified multiple cell lines that endogenously express gustatory signaling components and respond with the anticipated intracellular calcium and cAMP changes upon stimulation with tastants. These cell lines can be readily used for studying the gustatory cascade or identifying small molecules that modulate MCC through the gustatory signaling cascade."