Novel methods of applying direct chemical and mechanical stimulation to the oral mucosa for traditional behavioral pain assays in conscious rats
Novel methods of applying direct chemical and mechanical stimulation to the oral mucosa for traditional behavioral pain assays in conscious rats
复制标题
DOI：
10.1016/j.jneumeth.2014.10.013
复制
发表时间：
2015-01-15
影响因子：
3
通讯作者：
Inenaga, Kiyotoshi
中科院分区：
文献类型：
--
作者：
Hitomi, Suzuro;Ono, Kentaro;Inenaga, Kiyotoshi
背景：口腔炎引起严重和痛苦的过敏，对辛辣和身体接触在吃饭。许多研究使用麻醉动物来检查口腔粘膜中诱发的伤害感受，但没有报道使用传统的行为分析来评估清醒动物的伤害感受。新方法：我们开发了两种新方法，分别对有意识大鼠的下颌前庭口腔黏膜进行化学或机械刺激。通过测量面部梳理时间和头部对von Frey刺激的退缩阈值来进行伤害性评估。(1)口内滴注法，用手暂时暴露大鼠黏膜，滴入一滴刺激性溶液。(2)稳定口内开放法，在习惯训练2-3周后，长期暴露大鼠心理皮肤穿刺手术后的粘膜。结果：在口腔滴注法中，应用100 μ M辣椒素或100 μ M异硫氰酸烯丙酯可延长口腔摩擦时间。辣椒素诱导的口腔摩擦时间在醋酸诱导的溃疡发生后进一步延长。稳定口内开放法可以稳定地测量清醒大鼠口腔黏膜的机械戒断阈值。溃疡的发展降低了机械阈值，而局部利多卡因治疗增加了机械阈值。与现有方法的比较：这些新方法可以在没有任何麻醉作用的情况下评估口腔内刺激的动机性恶意行为。结论：口内滴注法和稳定口内开放法可与传统行为学方法相结合，用于评价清醒大鼠口腔黏膜的伤害感受。(C) 2014 Elsevier B.V.版权所有
Background: Stomatitis induces severe and painful hypersensitivity to pungency and physical contact during meals. Many studies have used anesthetized animals to examine evoked nociception in the oral mucosa, but no reports have used traditional behavioral assays to evaluate nociception in conscious animals.New methods: We developed two new methods of applying chemical or mechanical stimulation directly to the oral mucosa of the mandibular vestibule of conscious rats. Nociceptive evaluations were performed by measuring facial grooming time and the head withdrawal threshold to von Frey stimulations. (I) For the intraoral dropping method, rat mucosa was transiently exposed by hand, and a drop of a pungent solution was applied. (2) For the stable intraoral opening method, rat mucosa was long-term exposed following piercing surgery of the mental skin after habitual training for 2-3 weeks.Results: In the intraoral dropping method, the application of 100 mu M capsaicin or 100 mM allyl isothiocyanate prolonged mouth-rubbing time. Capsaicin-induced mouth-rubbing time was further enhanced following the development of an acetic acid-induced ulcer. The stable intraoral opening method enabled stable measurements of the mechanical withdrawal threshold in the oral mucosa of conscious rats. Ulcer development decreased the mechanical threshold, whereas topical lidocaine treatment increased the threshold.Comparison with existing methods: These new methods enable the evaluations of motivational nocifensive behaviors in response to intraoral stimulations without any anesthetic effects.Conclusions: The intraoral dropping and stable intraoral opening methods can be used in combination with traditional behavioral assays to evaluate nociception in the oral mucosa of conscious rats. (C) 2014 Elsevier B.V. All rights reserved.