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

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Abstract

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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Acknowledgements

This work was supported by a Discovery grant from the Natural Sciences and Engineering Research Council of Canada (RGPIN-2014-04099) and a Manitoba Medical Service Foundation Allen Rouse Career Award to PC, and a catalyst grant from the Biology of Breathing theme, Children’s Hospital Research Institute (CHRIM), to KD and PC. AJ and JDU were supported by graduate studentships from the University of Manitoba and Research Manitoba.

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Authors and Affiliations

  1. Manitoba Chemosensory Biology (MCSB) Research Group, University of Manitoba, Winnipeg, MB, R3E 0W4, Canada

    Appalaraju Jaggupilli, Nisha Singh, Jasbir Upadhyaya, Anurag S. Sikarwar, Makoto Arakawa, Shyamala Dakshinamurti, Rajinder P. Bhullar, Kangmin Duan & Prashen Chelikani

  2. Department of Oral Biology, University of Manitoba, Winnipeg, MB, R3E 0W4, Canada

    Appalaraju Jaggupilli, Nisha Singh, Jasbir Upadhyaya, Anurag S. Sikarwar, Makoto Arakawa, Rajinder P. Bhullar, Kangmin Duan & Prashen Chelikani

  3. Department of Dental Hygiene, Chiba Prefectural University of Health Sciences, Winnipeg, Canada

    Makoto Arakawa

  4. Departments of Pediatrics, Physiology, University of Manitoba, Winnipeg, Canada

    Shyamala Dakshinamurti

  5. Biology of Breathing, Children’s Hospital Research Institute of Manitoba (CHRIM), Winnipeg, Canada

    Shyamala Dakshinamurti, Kangmin Duan & Prashen Chelikani

  6. Department of Pharmacology and Therapeutics, University of Manitoba, Winnipeg, MB, R3E 0W4, Canada

    Prashen Chelikani

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  1. Appalaraju Jaggupilli
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Correspondence to Prashen Chelikani.

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Jaggupilli, A., Singh, N., Upadhyaya, J. et al. Analysis of the expression of human bitter taste receptors in extraoral tissues. Mol Cell Biochem 426, 137–147 (2017). https://doi.org/10.1007/s11010-016-2902-z

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