Analysis of the expression of human bitter taste receptors in extraoral tissues

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作者
Appalaraju Jaggupilli
Nisha Singh
Jasbir Upadhyaya
Anurag S. Sikarwar
Makoto Arakawa
Shyamala Dakshinamurti
Rajinder P. Bhullar
Kangmin Duan
Prashen Chelikani
机构
[1] University of Manitoba,Manitoba Chemosensory Biology (MCSB) Research Group
[2] University of Manitoba,Department of Oral Biology
[3] Chiba Prefectural University of Health Sciences,Department of Dental Hygiene
[4] University of Manitoba,Departments of Pediatrics, Physiology
[5] Children’s Hospital Research Institute of Manitoba (CHRIM),Biology of Breathing
[6] University of Manitoba,Department of Pharmacology and Therapeutics
来源
关键词
Bitter taste receptor gene (TAS2R); Bitter taste receptor protein (T2R); G protein-coupled receptor (GPCR); nCounter; gene sequencing; Cystic fibrosis bronchial epithelial cells (CuFi-1); Normal bronchial epithelial cells (NuLi-1); Airway smooth muscle cells (ASMCs); Breast cancer cells (MDA-MB-231);
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摘要
The 25 bitter taste receptors (T2Rs) in humans perform a chemosensory function. However, very little is known about the level of expression of these receptors in different tissues. In this study, using nCounter gene expression we analyzed the expression patterns of human TAS2R transcripts in cystic fibrosis bronchial epithelial (CuFi-1), normal bronchial epithelial (NuLi-1), airway smooth muscle (ASM), pulmonary artery smooth muscle (PASM), mammary epithelial, and breast cancer cells. Our results suggest a specific pattern of TAS2R expression with TAS2R3, 4, 5, 10, 13, 19, and 50 transcripts expressed at moderate levels and TAS2R14 and TAS2R20 (or TASR49) at high levels in the various tissues analyzed. This pattern of expression is mostly independent of tissue origin and the pathological state, except in cancer cells. To elucidate the expression at the protein level, we pursued flow cytometry analysis of select T2Rs from CuFi-1 and NuLi-1 cells. The expression levels observed at the gene level by nCounter analysis correlate with the protein levels for the T2Rs analyzed. Next, to assess the functionality of the expressed T2Rs in these cells, we pursued functional assays measuring intracellular calcium mobilization after stimulation with the bitter compound quinine. Using PLC inhibitor, U-73122, we show that the calcium mobilized in these cells predominantly takes place through the Quinine–T2R–Gαβγ–PLC pathway. This report will accelerate studies aimed at analyzing the pathophysiological function of T2Rs in different extraoral tissues.
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页码:137 / 147
页数:10
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