title: Functional expression of the extracellular-Ca2+-sensing receptor in mouse taste cells.
abstract: Three types of morphologically and functionally distinct taste cells operate in the mammalian taste bud. We demonstrate here the expression of two G-protein-coupled receptors from the family C, CASR and GPRC6A, in the taste tissue and identify transcripts for both receptors in type I cells, no transcripts in type II cells and only CASR transcripts in type III cells, by using the SMART-PCR RNA amplification method at the level of individual taste cells. Type I taste cells responded to calcimimetic NPS R-568, a stereoselective CASR probe, with Ca(2+) transients, whereas type I and type II cells were not specifically responsive. Consistent with these findings, certain amino acids stimulated PLC-dependent Ca(2+) signaling in type III cells, but not in type I and type II cells, showing the following order of efficacies: Phe~Glu>Arg. Thus, CASR is coupled to Ca(2+) mobilization solely in type III cells. CASR was cloned from the circumvallate papilla into a pIRES2-EGFP plasmid and heterologously expressed in HEK-293 cells. The transfection with CASR enabled HEK-293 cells to generate Ca(2+) transients in response to the amino acids, of which, Phe was most potent. This observation and some other facts favor CASR as the predominant receptor subtype endowing type III cells with the ability to detect amino acids. Altogether, our results indicate that type III cells can serve a novel chemosensory function by expressing the polymodal receptor CASR. A role for CASR and GPRC6A in physiology of taste cells of the type I remains to be unveiled.
authors.affiliation: Institute of Cell Biophysics, Russian Academy of Sciences, Institutional Street 3, Pushchino, Moscow Region, 142290, Russia.
authors.forename: Marina F
authors.initials: MF
authors.lastname: Bystrova
authors.forename: Roman A
authors.initials: RA
authors.lastname: Romanov
authors.forename: Olga A
authors.initials: OA
authors.lastname: Rogachevskaja
authors.forename: Gleb D
authors.initials: GD
authors.lastname: Churbanov
authors.forename: Stanislav S
authors.initials: SS
authors.lastname: Kolesnikov
mesh_terms: D000255:Adenosine Triphosphate / Q000494:pharmacology; D000596:Amino Acids / Q000494:pharmacology; D000814:Aniline Compounds / Q000494:pharmacology; D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002460:Cell Line; D005110:Extracellular Space / Q000187:drug effects / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D015640:Ion Channel Gating / Q000187:drug effects; D051379:Mice; D010627:Phenethylamines; D011189:Potassium Chloride / Q000494:pharmacology; D011437:Propylamines; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D044169:Receptors, Calcium-Sensing / Q000235:genetics / Q000378:metabolism*; D043562:Receptors, G-Protein-Coupled / Q000235:genetics / Q000378:metabolism; D013650:Taste Buds / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D014162:Transfection; D010738:Type C Phospholipases / Q000378:metabolism
chemical_list: D000596:Amino Acids; D000814:Aniline Compounds; C515750:GPRC6A protein, mouse; C107873:N-(2-chlorophenylpropyl)-1-(3-methoxyphenyl)ethylamine; D010627:Phenethylamines; D011437:Propylamines; D012333:RNA, Messenger; D044169:Receptors, Calcium-Sensing; D043562:Receptors, G-Protein-Coupled; D011189:Potassium Chloride; D000255:Adenosine Triphosphate; D010738:Type C Phospholipases; D002118:Calcium