TY - JOUR
T1 - Nicotinic acetylcholine receptor (CHRN) expression and function in cultured human adult fungiform (HBO) taste cells
AU - Qian, Jie
AU - Mummalaneni, Shobha
AU - Larsen, James
AU - Grider, John R.
AU - Spielman, Andrew I.
AU - Özdener, Mehmet Hakan
AU - Lyall, Vijay
N1 - Funding Information:
Supported by National Institute on Deafness and other Communication Disorders (NIDCD) grant DC-011569 (VL), Deans Bridge Funds from the VCU School of Medicine (VL), and DK34153 (JRG). Microscopy was performed at the VCU Department of Anatomy and Neurobiology Microscopy Facility, supported, in part, with funding from NIH-NINDS Center core grant (5P30NS047463). Some of the imaging experiments were performed at the Monell Chemical Senses Center “The Histology and Cellular Localization Core”, which is supported, in part, by funding from the NIH-NIDCD Core Grant P30DC011735.
Funding Information:
SupportedbyNationalInstituteonDeafnessandotherCommunicationDisorders(NIDCD) grantDC-011569(VL),DeansBridgeFundsfromtheVCUSchoolofMedicine(VL),and DK34153(JRG).MicroscopywasperformedattheVCUDepartmentofAnatomyandNeuro-biologyMicroscopyFacility,supported,inpart,withfundingfromNIH-NINDSCentercore grant(5P30NS047463).SomeoftheimagingexperimentswereperformedattheMonell ChemicalSensesCenter“TheHistologyandCellularLocalizationCore”,whichissupported, inpart,byfundingfromtheNIH-NIDCDCoreGrantP30DC011735.
Publisher Copyright:
© 2018 Qian et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
PY - 2018/3
Y1 - 2018/3
N2 - In rodents, CHRNs are involved in bitter taste transduction of nicotine and ethanol. Currently, it is not clear if CHRNs are expressed in human taste cells and if they play a role in transducing the bitter taste of nicotine and ethanol or in the synthesis and release of neuro-humoral peptides. Accordingly, we investigated the expression and functional role of CHRNs in HBO cells. Using molecular techniques, we demonstrate that a subset of HBO cells express CHRNs that also co-express TRPM5, T1R3 or T2R38. Exposing HBO cells to nicotine or ethanol acutely or to nicotine chronically induced a differential increase in the expression of CHRN mRNA and protein in a dose- and time-dependent manner. Acutely exposing HBO cells to a mixture containing nicotine plus ethanol induced a smaller increase in CHRN mRNAs relative to nicotine or ethanol treatment alone. A subset of HBO cells responded to nicotine, acetylcholine and ATP with a transient increase in [Ca2+]i. Nicotine effects on [Ca2+]i were mecamylamine sensitive. Brain-derived neurotrophic factor (BDNF) protein was detected in HBO cells using ELISA. Acute nicotine exposure decreased BDNF in HBO cells and increased BDNF release in the medium. CHRNs were also detected in HEK293 cells by RT-PCR. Unlike HBO cells, CHRNs were localized in most of HEK293 cells and majority of HEK293 cells responded to nicotine and ethanol stimulation with a transient increase in [Ca2+]i. BDNF levels in HEK293 cells were significantly higher than in HBO cells but the nicotine induced release of BDNF in the media was a fraction of the BDNF cellular content. We conclude that CHRNs are expressed in TRPM5 positive HBO cells. CHRN mRNA expression is modulated by exposure to nicotine and ethanol in a dose- and time-dependent manner. Nicotine induces the synthesis and release of BDNF in HBO cells.
AB - In rodents, CHRNs are involved in bitter taste transduction of nicotine and ethanol. Currently, it is not clear if CHRNs are expressed in human taste cells and if they play a role in transducing the bitter taste of nicotine and ethanol or in the synthesis and release of neuro-humoral peptides. Accordingly, we investigated the expression and functional role of CHRNs in HBO cells. Using molecular techniques, we demonstrate that a subset of HBO cells express CHRNs that also co-express TRPM5, T1R3 or T2R38. Exposing HBO cells to nicotine or ethanol acutely or to nicotine chronically induced a differential increase in the expression of CHRN mRNA and protein in a dose- and time-dependent manner. Acutely exposing HBO cells to a mixture containing nicotine plus ethanol induced a smaller increase in CHRN mRNAs relative to nicotine or ethanol treatment alone. A subset of HBO cells responded to nicotine, acetylcholine and ATP with a transient increase in [Ca2+]i. Nicotine effects on [Ca2+]i were mecamylamine sensitive. Brain-derived neurotrophic factor (BDNF) protein was detected in HBO cells using ELISA. Acute nicotine exposure decreased BDNF in HBO cells and increased BDNF release in the medium. CHRNs were also detected in HEK293 cells by RT-PCR. Unlike HBO cells, CHRNs were localized in most of HEK293 cells and majority of HEK293 cells responded to nicotine and ethanol stimulation with a transient increase in [Ca2+]i. BDNF levels in HEK293 cells were significantly higher than in HBO cells but the nicotine induced release of BDNF in the media was a fraction of the BDNF cellular content. We conclude that CHRNs are expressed in TRPM5 positive HBO cells. CHRN mRNA expression is modulated by exposure to nicotine and ethanol in a dose- and time-dependent manner. Nicotine induces the synthesis and release of BDNF in HBO cells.
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U2 - 10.1371/journal.pone.0194089
DO - 10.1371/journal.pone.0194089
M3 - Article
C2 - 29513745
AN - SCOPUS:85042905045
SN - 1932-6203
VL - 13
JO - PloS one
JF - PloS one
IS - 3
M1 - e0194089
ER -