Action potential-dependent ATP release from taste receptor cells : development of real-time micro determination for neurotransmitter release at single ceII level

味觉受体细胞的动作电位依赖性 ATP 释放：单细胞水平神经递质释放实时微量测定的开发

• 批准号: 20791355
• 负责人: MURATA Yoshihiro
• 金额: $ 2.58万
• 依托单位: Kyushu University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Young Scientists (B)
• 财政年份: 2008
• 资助国家: 日本
• 起止时间: 2008 至 2009
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-20791355/
• 关键词: 味細胞; 活動電位; シナプス; 神経伝達物質; ATP; ヘミチャネル

Subsets of taste bud cells generate action potentials in response to sapid stimuli. Type II taste cells have many taste transduction molecules but lack well elaborated synapses, which leads to the question in the functional significance of action potentials in these cells. We examined the dependence of adenosine triphosphate (ATP) transmitter release from taste cells upon action potentials. To identify Type TT taste cells we used mice expressing a green fluorescence protein (GFP) transgene from the gustducin promoter. Action potentials were recorded by an electrode basolaterally attached to a. single GFP-positive taste cell. ATP release was monitored from gustducin-expressing taste cells by collecting the electrode solution immediately after tastant-stimulated action potentials and using a. luciferase assay to quantify ATP. Stimulation of gustducin-expressing taste cells with a. sweet, bitter, or umami compound on the apical membrane increased ATP levels in the electrode solution; the amount of ATP depended on the firing rate. Increased spontaneous firing rates also induced ATP release from gustducin-expressing taste cells. ATP release from gustducin-expressing taste cells was depressed by tetrodotoxin and inhibited below the detection limit by carbenoxolone. Our data support the hypothesis that action potentials in Type II taste cells enhance ATP release through hemichannels.

味蕾细胞的亚群对味觉刺激产生动作电位。II型味觉细胞具有许多味觉传导分子，但缺乏精细的突触，这导致了这些细胞中动作电位的功能意义的问题。我们研究了三磷酸腺苷（ATP）递质释放的依赖性味觉细胞动作电位。为了鉴定TT型味觉细胞，我们使用了表达来自gustducin启动子的绿色荧光蛋白（GFP）转基因的小鼠。动作电位记录的电极基底侧连接到一个。单个GFP阳性味觉细胞。通过在味素刺激的动作电位后立即收集电极溶液并使用a.荧光素酶测定以定量ATP。用a.顶端膜上的甜味、苦味或鲜味化合物增加了电极溶液中的ATP水平; ATP的量取决于放电速率。增加自发放电率也诱导ATP从表达味蛋白的味觉细胞释放。河豚毒素抑制表达味细胞的ATP释放，甘珀酸抑制低于检测限。我们的数据支持这一假设，即在II型味觉细胞的动作电位增强ATP释放通过半通道。

项目成果

活動電位を発生する味細胞からのATP放出におけるヘミチャネルの関与

半通道参与味觉细胞释放 ATP 并产生动作电位

• 发表时间: 2008
• 作者: 村田芳博;吉田竜介;安尾敏明;柳川右千夫;小幡邦彦;植野洋志;Margolskee RF;二ノ宮裕三
• 通讯作者: 二ノ宮裕三

Multiple receptor systems for umami taste in mice

小鼠鲜味的多重受体系统

• 发表时间: 2008
• 作者: Yoshida R;Yasumatsu K;Shirosaki S;Kawato Y;Murata Y;Shigemura N;Nakashima K;Margolskee RF;Ninomiya Y (Invited speaker)
• 通讯作者: Ninomiya Y (Invited speaker)

甘味の受容・伝達・調節機構とATP

甜味接收、传递和调节机制与 ATP

• 发表时间: 2008
• 作者: 吉田竜介、村田芳博、安松啓子;重村憲徳;二ノ宮裕三
• 通讯作者: 二ノ宮裕三

NaCl RESPONSIVE TASTE CELLS IN THE MOUSE FUNGIFORM TASTE BUDS

• DOI: 10.1016/j.neuroscience.2008.12.052
• 发表时间: 2009-03-17
• 期刊: NEUROSCIENCE
• 影响因子: 3.3
• 作者: Yoshida, R.;Horio, N.;Ninomiya, Y.
• 通讯作者: Ninomiya, Y.

Action potential-dependent ATP release from Type II taste cells through hemichannels in mice

小鼠 II 型味觉细胞通过半通道释放动作电位依赖性 ATP

• 发表时间: 2009
• 作者: Murata Y;Yasuo T;Yoshida R;Obata K;Yanagawa Y;Margolskee RF;Ninomiya Y
• 通讯作者: Ninomiya Y