Vandenbeuch Aurelie, Clapp Tod R, Kinnamon Sue C
Department of Biomedical Science, Colorado State University, Fort Collins, USA.
BMC Neurosci. 2008 Jan 2;9:1. doi: 10.1186/1471-2202-9-1.
Taste buds are the sensory organs of taste perception. Three types of taste cells have been described. Type I cells have voltage-gated outward currents, but lack voltage-gated inward currents. These cells have been presumed to play only a support role in the taste bud. Type II cells have voltage-gated Na+ and K+ current, and the receptors and transduction machinery for bitter, sweet, and umami taste stimuli. Type III cells have voltage-gated Na+, K+, and Ca2+ currents, and make prominent synapses with afferent nerve fibers. Na+ salt transduction in part involves amiloride-sensitive epithelial sodium channels (ENaCs). In rodents, these channels are located in taste cells of fungiform papillae on the anterior part of the tongue innervated by the chorda tympani nerve. However, the taste cell type that expresses ENaCs is not known. This study used whole cell recordings of single fungiform taste cells of transgenic mice expressing GFP in Type II taste cells to identify the taste cells responding to amiloride. We also used immunocytochemistry to further define and compare cell types in fungiform and circumvallate taste buds of these mice.
Taste cell types were identified by their response to depolarizing voltage steps and their presence or absence of GFP fluorescence. TRPM5-GFP taste cells expressed large voltage-gated Na+ and K+ currents, but lacked voltage-gated Ca2+ currents, as expected from previous studies. Approximately half of the unlabeled cells had similar membrane properties, suggesting they comprise a separate population of Type II cells. The other half expressed voltage-gated outward currents only, typical of Type I cells. A single taste cell had voltage-gated Ca2+ current characteristic of Type III cells. Responses to amiloride occurred only in cells that lacked voltage-gated inward currents. Immunocytochemistry showed that fungiform taste buds have significantly fewer Type II cells expressing PLC signalling components, and significantly fewer Type III cells than circumvallate taste buds.
The principal finding is that amiloride-sensitive Na+ channels appear to be expressed in cells that lack voltage-gated inward currents, likely the Type I taste cells. These cells were previously assumed to provide only a support function in the taste bud.
味蕾是味觉感知的感觉器官。已描述了三种类型的味觉细胞。I 型细胞具有电压门控外向电流,但缺乏电压门控内向电流。这些细胞被认为在味蕾中仅起支持作用。II 型细胞具有电压门控的 Na⁺和 K⁺电流,以及苦味、甜味和鲜味味觉刺激的受体和转导机制。III 型细胞具有电压门控的 Na⁺、K⁺和 Ca²⁺电流,并与传入神经纤维形成明显的突触。Na⁺盐转导部分涉及amiloride 敏感的上皮钠通道(ENaC)。在啮齿动物中,这些通道位于由鼓索神经支配的舌前部菌状乳头的味觉细胞中。然而,表达 ENaC 的味觉细胞类型尚不清楚。本研究使用在 II 型味觉细胞中表达 GFP 的转基因小鼠的单个菌状味觉细胞的全细胞记录来鉴定对 amiloride 有反应的味觉细胞。我们还使用免疫细胞化学来进一步定义和比较这些小鼠的菌状和轮廓乳头味蕾中的细胞类型。
通过对去极化电压阶跃的反应以及 GFP 荧光的有无来鉴定味觉细胞类型。TRPM5-GFP 味觉细胞表达大的电压门控 Na⁺和 K⁺电流,但缺乏电压门控 Ca²⁺电流,正如先前研究所预期的那样。大约一半未标记的细胞具有相似的膜特性,表明它们构成了一个单独的 II 型细胞群体。另一半仅表达电压门控外向电流,这是 I 型细胞的典型特征。单个味觉细胞具有 III 型细胞特有的电压门控 Ca²⁺电流。对 amiloride 的反应仅发生在缺乏电压门控内向电流的细胞中。免疫细胞化学显示,菌状味蕾中表达 PLC 信号成分的 II 型细胞明显少于轮廓乳头味蕾,且 III 型细胞也明显少于轮廓乳头味蕾。
主要发现是 amiloride 敏感的 Na⁺通道似乎在缺乏电压门控内向电流的细胞中表达,可能是 I 型味觉细胞。这些细胞以前被认为在味蕾中仅提供支持功能。