Dept. of Physiology and Biophysics, Virginia Commonwealth Univ., Molecular Medical Research Bldg. 5052, 1220 East Broad St., Richmond, VA 23298, USA.
J Neurophysiol. 2010 Mar;103(3):1337-49. doi: 10.1152/jn.00883.2009. Epub 2009 Dec 23.
Regulation of the putative amiloride and benzamil (Bz)-insensitive TRPV1t salt taste receptor by phosphatidylinositol 4,5-bisphosphate (PIP(2)) was studied by monitoring chorda tympani (CT) taste nerve responses to 0.1 M NaCl solutions containing Bz (5 x 10(-6) M; a specific ENaC blocker) and resiniferatoxin (RTX; 0-10 x 10(-6) M; a specific TRPV1 agonist) in Sprague-Dawley rats and in wildtype (WT) and TRPV1 knockout (KO) mice. In rats and WT mice, RTX elicited a biphasic effect on the NaCl + Bz CT response, increasing the CT response between 0.25 x 10(-6) and 1 x 10(-6) M. At concentrations >1 x 10(-6) M, RTX inhibited the CT response. An increase in PIP(2) by topical lingual application of U73122 (a phospholipase C blocker) or diC8-PIP(2) (a short chain synthetic PIP(2)) inhibited the control NaCl + Bz CT response and decreased its sensitivity to RTX. A decrease in PIP(2) by topical lingual application of phenylarsine oxide (a phosphoinositide 4 kinase blocker) enhanced the control NaCl + Bz CT response, increased its sensitivity to RTX stimulation, and inhibited the desensitization of the CT response at RTX concentrations >1 x 10(-6) M. The ENaC-dependent NaCl CT responses were not altered by changes in PIP(2). An increase in PIP(2) enhanced CT responses to sweet (0.3 M sucrose) and bitter (0.01 M quinine) stimuli. RTX produced the same increase in the Bz-insensitive Na(+) response when present in salt solutions containing 0.1 M NaCl + Bz, 0.1 M monosodium glutamate + Bz, 0.1 M NaCl + Bz + 0.005 M SC45647, or 0.1 M NaCl + Bz + 0.01 M quinine. No effect of RTX was observed on CT responses in WT mice and rats in the presence of the TRPV1 blocker N-(3-methoxyphenyl)-4-chlorocinnamide (1 x 10(-6) M) or in TRPV1 KO mice. We conclude that PIP(2) is a common intracellular effector for sweet, bitter, umami, and TRPV1t-dependent salt taste, although in the last case, PIP(2) seems to directly regulate the taste receptor protein itself, i.e., the TRPV1 ion channel or its taste receptor variant, TRPV1t.
研究了二磷酸酰肌醇 4,5-二磷酸(PIP(2))对假定的阿米洛利和苯甲脒(Bz)不敏感 TRPV1t 盐味觉受体的调节作用,方法是监测氯化物听觉(CT)味觉神经对含有 Bz(5×10(-6)M;一种特定的 ENaC 阻断剂)和树脂毒素(RTX;0-10×10(-6)M;一种特定的 TRPV1 激动剂)的 0.1M NaCl 溶液的反应,在 Sprague-Dawley 大鼠以及野生型(WT)和 TRPV1 敲除(KO)小鼠中。在大鼠和 WT 小鼠中,RTX 对 NaCl+Bz CT 反应产生双相作用,在 0.25×10(-6)和 1×10(-6)M 之间增加 CT 反应。在浓度>1×10(-6)M 时,RTX 抑制 CT 反应。通过局部舌应用 U73122(一种磷脂酶 C 阻断剂)或二 C8-PIP(2)(一种短链合成 PIP(2))增加 PIP(2)抑制对照 NaCl+Bz CT 反应,并降低其对 RTX 的敏感性。通过局部舌应用苯砷氧化物(一种磷酸肌醇 4 激酶阻断剂)降低 PIP(2)减少了对照 NaCl+Bz CT 反应,增加了其对 RTX 刺激的敏感性,并抑制了 RTX 浓度>1×10(-6)M 时 CT 反应的脱敏作用。ENaC 依赖性 NaCl CT 反应不受 PIP(2)变化的影响。PIP(2)的增加增强了对甜味(0.3M 蔗糖)和苦味(0.01M 奎宁)刺激的 CT 反应。当 RTX 存在于含有 0.1M NaCl+Bz、0.1M 单谷氨酸钠+Bz、0.1M NaCl+Bz+0.005M SC45647 或 0.1M NaCl+Bz+0.01M 奎宁的盐溶液中时,它对 Bz 不敏感的 Na(+)反应产生相同的增加。在 WT 小鼠和大鼠存在 TRPV1 阻断剂 N-(3-甲氧基苯基)-4-氯肉桂酰胺(1×10(-6)M)或 TRPV1 KO 小鼠的情况下,未观察到 RTX 对 CT 反应的影响。我们得出结论,PIP(2)是甜味、苦味、鲜味和 TRPV1t 依赖性盐味觉的共同细胞内效应物,尽管在后一种情况下,PIP(2)似乎直接调节味觉受体蛋白本身,即 TRPV1 离子通道或其味觉受体变体 TRPV1t。