Regulation of the benzamil-insensitive salt taste receptor by intracellular Ca2+, protein kinase C, and calcineurin.
The regulation of the benzamil (Bz)-insensitive salt taste receptor was investigated by intracellular Ca2+ ([Ca2+]i), protein kinase C (PKC), and the Ca2+-dependent serine-threonine phosphatase, calcineurin (PP2B), by monitoring chorda tympani taste nerve responses to 0.1 M NaCl solutions containing Bz (5x10(-6) M) and resiniferatoxin (RTX; 0-10x10(-6) M) in Sprague-Dawley rats and in wild-type (WT) and transient receptor potential vanilloid-1 knockout (TRPV1 KO) mice. In rats and WT mice, RTX increased the NaCl+Bz chorda tympani responses between 0.25x10(-6) and 1x10(-6) M and inhibited the responses above 1x10(-6) M. Decreasing taste receptor cell (TRC) [Ca2+]i with BAPTA loading, activation of PKC with 4alpha-phorbol-12,13-didecanoate (PMA), or inhibition of PP2B by cyclosporin A or FK-506, enhanced the magnitude of the Bz-insensitive NaCl chorda tympani responses in the presence of RTX and either minimized or completely eliminated the decrease in the chorda tympani response>1x10(-6) M RTX. In contrast, increasing TRC [Ca2+]i with ionomycin inhibited Bz-insensitive NaCl chorda tympani responses in the presence of RTX. No effect of the cited modulators was observed on the chorda tympani responses in WT mice and rats in the presence of TRPV1 blocker SB-366791 (1x10(-6) M) or in TRPV1 KO mice. 32P-labeling demonstrated direct phosphorylation of TRPV1 or TRPV1t in anterior lingual epithelium by PMA, cyclosporin A, or FK-506. PMA also enhanced the RTX-sensitive unilateral apical Na+ flux in polarized fungiform TRC in vitro. We conclude that TRPV1 or its variant TRPV1t is phosphorylated and dephosphorylated by PKC and PP2B, respectively, and either sensitizes or desensitizes the Bz-insensitive NaCl chorda tympani responses to RTX stimulation.
Lyall
Vijay
V
Department of Physiology and Biophysics, Virginia Commonwealth University, Sanger Hall, Room 3-010, 1101 E. Marshall St., Richmond, VA 23298-0551, USA. vlyall@vcu.edu
Phan
Tam-Hao T
TH
Mummalaneni
Shobha
S
Melone
Pamela
P
Mahavadi
Sunila
S
Murthy
Karnam S
KS
DeSimone
John A
JA
D000584:Amiloride / Q000031:analogs & derivatives; D000818:Animals; D001549:Benzamides / Q000494:pharmacology; D019703:Calcineurin / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism*; D002614:Chelating Agents / Q000494:pharmacology; D002814:Chorda Tympani Nerve / Q000166:cytology / Q000502:physiology; D004224:Diterpenes / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004533:Egtazic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D045604:Extracellular Fluid / Q000187:drug effects / Q000378:metabolism*; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects / Q000235:genetics; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D010704:Phorbols / Q000494:pharmacology; D011493:Protein Kinase C / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012965:Sodium Chloride / Q000494:pharmacology; D050916:TRPV Cation Channels / Q000037:antagonists & inhibitors / Q000172:deficiency; D013649:Taste / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D013650:Taste Buds / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D013652:Taste Threshold / Q000187:drug effects / Q000235:genetics
D001549:Benzamides; D002614:Chelating Agents; D004224:Diterpenes; D004791:Enzyme Inhibitors; D010704:Phorbols; D050916:TRPV Cation Channels; C493921:TRPV1 protein, mouse; C013407:benzamil; D012965:Sodium Chloride; D004533:Egtazic Acid; C037689:benzamide; D000584:Amiloride; C024353:resiniferatoxin; D011493:Protein Kinase C; D019703:Calcineurin; C025603:1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid; D002118:Calcium; C033085:phorbol