Role of α_2-adrenoreceptor on the excitability of trigeminal root ganglion neurons : Patch-clamp and Single-cell RT-PCR analysis

α_2-肾上腺素受体对三叉神经根神经节神经元兴奋性的作用：膜片钳和单细胞 RT-PCR 分析

• 批准号: 13671953
• 负责人: TAKEDA Mamoru
• 金额: $ 1.92万
• 依托单位: Nippon Dental University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13671953/
• 关键词: trigeminal root ganglion; perforated patch-clamp; Reverse transcription-Polymerase chain reaction; α_2-adrenoreceptors; I_h; Whole-cell patch-clamp; nociception; Single-cell; 穿孔パッチ法; I_h電流; クロニジン

The aim of the study was to mauiine the effects of α_2-adrerioreceptar agonists on the excitability of teigeminalroot ganglion (TRG) neurons using the perforated-patch damp technique, and to deternine wither these neurons express mRNA for α_2-adrenoreceptors. In cunent-clamp mode, the testing membrane potential was -57.4 ± 1.2 mV(n=26). Most neurons (71%) were hyperpolarized by clonidine (5-50μM) in a concentration-dependent manner. The response was associated with an increase of cell input resistance. In addition, clonidine reduced the repetitive firing evoked by depolarizing current pulses. An α_2-adrenergic agonist UK14,304 (10-20μM) also hyperpolarized TRG neurons. The donidine- and UK14, 304- induced hyperpdarizatian was blocked by idazoxan (α_2-adrenoreoeptor antagonist). In voltage-damp, donidine (1-50μM) reversibly reduced the hyperpolarization- and -time dependent cationic current. The effect was mimicked by UK13,304 (10-20μM), and antagonized by idazoxan. Hyperpoiarization-activated cationic current was blocked by extracelluar Cs^+ (2mM) or a specific blocker, ZD7288 (20μM). Analysis of taii-cinrcnts revealed that a reversal potential of the clonidine sensitive component of hyperpolarization-activated cationic current was -46mV. Single cell reverse transcription-polymerase chain reaction analysis demonstrated the expression of mRNA for α_<2A> and α_<2C> adrenoreceptors.These results demonstrate that activation of α_2-adrenoreceptors can hyperpolarize TRG neurons, and that the in hibitory effect is associated with inhibition of hyperpolarization-activated cationic current. Our results suggest that activation of α_2-adrenoreceptors in the absence of nerve injury may have an inhibitory effect on nociceptive transmission in the trigeminal system at the level of both TRG neuronal cell bodies and primary afferent terminals.

该研究的目的是使用穿孔贴片潮湿技术来研究α_2-肾上腺素受体激动剂对四叉根神经节（TRG）神经元兴奋性的影响，并确定这些神经元是否表达α_2-肾上腺素受体的mRNA。电流钳模式下，测试膜电位为-57.4±1.2 mV(n=26)。大多数神经元（71%）被可乐定（5-50μM）以浓度依赖性方式超极化。该反应与细胞输入电阻的增加有关。此外，可乐定还减少了去极化电流脉冲引起的重复放电。 α_2-肾上腺素激动剂 UK14,304 (10-20μM) 也使 TRG 神经元超极化。多尼定和 UK14, 304 诱导的 hyperpdarizatian 被咪唑克生（α_2-肾上腺素受体拮抗剂）阻断。在电压潮湿的情况下，多尼丁 (1-50μM) 可逆地降低超极化和时间依赖性阳离子电流。 UK13,304 (10-20μM) 模仿了该作用，并被咪唑克生拮抗。超极化激活的阳离子电流被细胞外 Cs^+ (2mM) 或特定阻断剂 ZD7288 (20μM) 阻断。尾电流分析表明，超极化激活的阳离子电流可乐定敏感成分的反转电位为-46mV。单细胞逆转录聚合酶链反应分析表明α_2A和α_2C>肾上腺素受体mRNA的表达。这些结果表明α_2-肾上腺素受体的激活可以使TRG神经元超极化，并且抑制效应与超极化激活的阳离子电流的抑制有关。我们的结果表明，在没有神经损伤的情况下，α_2-肾上腺素受体的激活可能对三叉神经系统中 TRG 神经元细胞体和初级传入末梢水平的伤害性传递具有抑制作用。

项目成果

M. Takeda, M. Ikeda, T. Tanimoto, J. Lipski, S.Matsumoto: "Changes of excitability of rat trigeminal ganglion neurons evoked by α_2-adrenoreceptors"Neuroscience. 115. 731-741 (2002)

M. Takeda、M. Ikeda、T. Tanimoto、J. Lipski、S. Matsumoto：“α_2-肾上腺素受体诱发的大鼠三叉神经节神经元的兴奋性变化”神经科学。 115. 731-741 (2002)

M.Takeda, M.Ikeda, T.Tanimoto, J.Lipski, S.Matsumoto: "Changes of excitability of rat trigeminal ganglion neurons evoked by α_2-adrenoreceptors"Neuroscience. 115. 731-741 (2002)

M.Takeda、M.Ikeda、T.Tanimoto、J.Lipski、S.Matsumoto：“α_2-肾上腺素受体诱发的大鼠三叉神经节神经元的兴奋性变化”神经科学 115. 731-741 (2002)。

M.Takeda, M.Ikeda, T.Tanimoto, J.Lipski, S.Matsumoto: "Changes of excitability of rat trigeminal ganglion neurons evoked by α2-adrenoreceptors"Neuroscience. 115. 731-741 (2002)

M.Takeda、M.Ikeda、T.Tanimoto、J.Lipski、S.Matsumoto：“α2-肾上腺素受体诱发的大鼠三叉神经节神经元的兴奋性变化”神经科学 115. 731-741 (2002)。