Studies for diversity of modulation system on Pain and Capsaicin sensitivities

疼痛和辣椒素敏感性调节系统多样性的研究

• 批准号: 16300138
• 负责人: KOIDE Tsuyoshi
• 金额: $ 9.6万
• 依托单位: National Institute of Genetics
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-16300138/
• 关键词: Pain; Capsaicin; TRP-V1; Mouse; Congenic strains; Thermo sensitivity

In animal, heat stimulation is transmitted to central nervous system (CNS) via sensory neuron, and is recognized as heat or pain. Various levels of heat sensations activate the multi functional pain receptors, TRPV channels that expressed in sensory ganglia. Many behavioral tests had been established to measure the pain sensitivity in rodents by investigating behavioral response from the aversive pain stimuli. Hot plate test displays the ability of pain recognition system or analgesic system based on upper CNS. The tail flick test displays the pain sensitivity based on nociceptive reflex. In the present study, we conducted QTL analyses to characterize genetic loci involved in the different sensitivity to heat evoked pain between two mouse strains, C57BL/6 and KJR. In the study, we applied hot plate test and tail flick test to assess the two types of pain sensitivity of BKF2 progeny generated form F1 progeny of C57BL/6 and KJR. In the analyses, we detected three significant loci for hot … More plate test, Hpq1, Hpq2 and Hpq3 and two significant loci for tail flick test, Tfq1 and Tfq2. The pain associated quantitative traits loci, Hpq1, Hpq2, Hpq3, Tfq1, and Tfq2 were located on the chromosomes, 2,10,18,8 and 10, respectively. Hpq2 and Tfq1 were overlapped on the same region of chromosome 10. These data suggested that genetic mechanism for strain difference of pain sensitivities between C57BL/6 and KJR that mediated by upper CNS and reflection system are mostly different but partly shared.To elucidate phenotype of thermo-sensation, we conducted recording of femoral nerve activity in vivo and intracellular free calcium (iCa) response to heat in vitro in mice of KJR. KJR is the mice strain known to be resistant to hot-plate test. Increment in neuronal activity from 25 C to 40 C was not different in KJR compared to the control. Whereas that from 25 C to 50 C was was different in KJR, when the degree of the increment was compared with C57BL/6 by two-way ANOVA. DRG neurons were isolated from these mice and iCa was measured by ratio methods with fluo-3 and fura-red. Surprisingly, all of the phenotypes including capsaicin resistance in KJR are not reproduced in isolated DRG neurons : We observed rise in iCa in some DRG neurons of KJR to heat and capsaicin. These results may suggest that thermo-sensation is a complicated sensory process involving contribution of proteins other than TRP channels and that the experiments in vitro and in vivo are needed to substantiate the phenotype. Less

在动物中，热刺激通过感觉神经元传递到中枢神经系统（CNS），并被识别为热或疼痛。许多行为测试显示，不同程度的热感觉会激活多功能疼痛受体（TRPV 通道）。已建立通过研究厌恶性疼痛刺激的行为反应来测量啮齿类动物的疼痛敏感性。热板测试显示基于上中枢神经系统的疼痛识别系统或镇痛系统的能力。甩尾测试显示基于伤害性反射的疼痛敏感性。 。在本研究中，我们进行了 QTL 分析，以表征两种小鼠品系 C57BL/6 和 KJR 之间对热诱发疼痛的不同敏感性所涉及的遗传位点。在这项研究中，我们应用热板试验和甩尾试验来评估这两种小鼠。从 C57BL/6 和 KJR 的 F1 后代产生的 BKF2 后代的疼痛敏感性类型 在分析中，我们检测到热板测试的三个显着位点：Hpq1、Hpq2 和Hpq3 和甩尾测试的两个显着基因座 Tfq1 和 Tfq2 疼痛相关数量性状基因座 Hpq1、Hpq2、Hpq3、Tfq1 和 Tfq2 分别位于 2、10、18、8 和 10 号染色体上。和Tfq1在10号染色体的同一区域重叠。这些数据表明疼痛应变差异的遗传机制C57BL/6 和 KJR 之间由上 CNS 和反射系统介导的敏感性大部分不同，但部分相同。为了阐明热感觉的表型，我们记录了体内股神经活动和细胞内游离钙 (iCa) 对热的反应。 KJR 小鼠品系已知对 25°C 至 40°C 的神经元活性增加没有差异。 KJR 与对照相比，除了从 25°C 到 50°C 不同之外，当通过双向 ANOVA 与 C57BL/6 比较增量程度时，从这些小鼠中分离 DRG 神经元并通过测量 iCa。令人惊讶的是，KJR 中的所有表型（包括辣椒素抗性）均未在分离的 DRG 神经元中重现：我们观察到某些神经元中 iCa 升高。 KJR 的 DRG 神经元对热和辣椒素的影响可能表明热感觉是一个复杂的感觉过程，涉及 TRP 通道以外的蛋白质的贡献，并且需要体外和体内实验来证实该表型。

项目成果

Monoallelic yet combinatorial expression of variable exons of the protocadherin- a gene cluster in single neurons

原钙粘蛋白基因簇可变外显子在单个神经元中的单等位基因和组合表达

• 发表时间: 2005
• 期刊: Nature Genetics 37
• 作者: Ogasawara;M.;Imanishi;T.;Moriwaki;K.;Gaudieri;S.;Tsuda;H.;Hashimoto;H.;Shiroishi;T.;Gojobori;T.;Koide;T.;Esumi et al.
• 通讯作者: Esumi et al.

Molecular evolution of Cadherin-related neuronal receptor/protocadherin ・(CNR/Pcdh・) gene cluster in Mus musculus subspecies

小家鼠亚种钙粘蛋白相关神经元受体/原钙粘蛋白·(CNR/Pcdh·)基因簇的分子进化

• 期刊: Mol.Biol.Evol. (In press)
• 作者: Ogasawara;M.;Imanishi;T.;Moriwaki;K.;Gaudieri;S.;Tsuda;H.;Hashimoto;H.;Shiroishi;T.;Gojobori;T.;Koide;T.;Esumi et al.;Taguchi et al.
• 通讯作者: Taguchi et al.

カプサイシン受容体の活性調節剤、及びそれを用いた医薬組成物

辣椒素受体活性调节剂及使用其的药物组合物

• 发表时间: 2005