Characterization of Bitter Taste Receptor-Dependent Autophagy in Oral Epithelial Cells

Nisha Singh; Saeid Ghavami; Prashen Chelikani

doi:10.1007/7651_2024_531

Characterization of Bitter Taste Receptor-Dependent Autophagy in Oral Epithelial Cells

Methods Mol Biol. 2024 Apr 6. doi: 10.1007/7651_2024_531. Online ahead of print.

Authors

Nisha Singh^{1

2}, Saeid Ghavami^{3

4}, Prashen Chelikani^{5

6

7}

Affiliations

¹ Manitoba Chemosensory Biology (MCSB) Research Group, Winnipeg, MB, Canada.
² Department of Oral Biology, Dr. Gerald Niznick College of Dentistry, Rady Faculty of Health Sciences, University of Manitoba, Winnipeg, MB, Canada.
³ Research Institute of Oncology and Hematology, Cancer Care Manitoba-University of Manitoba, Winnipeg, MB, Canada. Saeid.ghavami@umanitoba.ca.
⁴ Department of Human Anatomy and Cell Science, University of Manitoba College of Medicine, Winnipeg, MB, Canada. Saeid.ghavami@umanitoba.ca.
⁵ Manitoba Chemosensory Biology (MCSB) Research Group, Winnipeg, MB, Canada. Prashen.Chelikani@umanitoba.ca.
⁶ Department of Oral Biology, Dr. Gerald Niznick College of Dentistry, Rady Faculty of Health Sciences, University of Manitoba, Winnipeg, MB, Canada. Prashen.Chelikani@umanitoba.ca.
⁷ Department of Biochemistry and Medical Genetics, Department of Physiology and Pathophysiology, Max Rady College of Medicine, Rady Faculty of Health Sciences, University of Manitoba, Winnipeg, MB, Canada. Prashen.Chelikani@umanitoba.ca.

PMID: 38578576
DOI: 10.1007/7651_2024_531

Abstract

Microbial dysbiosis is an important trigger in the development of oral diseases. Oral keratinocytes or gingival epithelial cells (GECs) offer protection against various microbial insults. Recent studies suggest that GECs expressed higher level of bitter taste receptor 14 (T2R14) compared to other taste receptors and toll-like receptors and act as innate immune sentinels. Macroautophagy or autophagy is a cellular conserved process involved in the regulation of host innate immune responses against microbial infection. Here, we describe a robust method for evaluation of T2R14-dependent autophagy flux in GECs. Autophagy flux was detected using Western blot analysis in GECs and further was confirmed using Acridine Orange-dependent flow cytometry analysis.

Keywords: Acridine Orange; Autophagosome; Autophagy flux inhibition; Bitter taste receptor; Flow cytometry; Oral epithelial cells; Serum starvation; Western blot.