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Comprehensive study on the plasticity and regeneration of the nucleus ambiguus motoneurons

疑核运动神经元可塑性与再生的综合研究

基本信息

批准号：
13470361
负责人：
HISA Yauso
金额：
$ 8.06万
依托单位：
Kyoto Prefectural University of Medicine
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2003
项目状态：
已结题

项目摘要

Molecular changes in neurons of rat nucleus ambiguus after axotomyNucleus ambiguus (NA) neurons projecting to intrinsic laryngeal muscles were labeled retrogradely with DiI by injecting into bilateral thyroarytenoid muscles, then either one of recurrent laryngeal nerves was transected, and fluorescence-bright single neurons were microdissected, followed by a single-cell reverse transcription (RT)-PCR. Immunohistochemically, many DiI-labeled motoneurons in NA were immunoreactive to phosphorylated Stat3 in the nucleus on 1,7,10 and 14 days after axotomy. Thirty □m-thick frozen sections of brain stem were mounted onto membrane slide, and DiI-labeled single neurons in NA were microdissected under a fluorescence microscope. By real-time quantitative RT-PCR for single motoneurons, the expression levels of GAP-43 and nNOS genes were up-regulated at 7 days and 1 day after axotomy, respectively. The expression levels of Stat3,Reg-2, and Bcl-2 genes were up-regulated at 7 days, whereas that of B … More ax was down-regulated at 1 and 7 days after axotomy as compared with control levels. We could demonstrate a part of molecular cascades involving Stat3 in neurons projecting to laryngeal muscles after axotomy with this novel method in which single-cell RT-PCR was combined with neurotracer DiI labeling.Distribution of vanilloid receptors in the rat laryngeal innervationWe investigated the distribution of vanilloid receptors in the rat larynx. VRL-1-positive fibers were detected in the laryngeal epithelium and lamina propria. VR1-positive nerve fibers were seen in the lamina propria but not in the mucosal epithelium. VR1- and VRL-1-positive cells were distributed in the intralaryngeal ganglia and colocalization of capsaicin receptors with VIP, nNOS and cChAT was seen. These findings suggest that these capsaicin receptors participate in the parasympathetic innervation as well as in nociception of the rat larynx.Expression of α-gustducin on taste buds in the rat larynxWe investigated the expression and distribution of α-gustducin in the taste buds of the rat larynx. The α-gustducin-positive taste buds were distributed most densely close to the caudal base of the laryngeal surface of the epiglottis, and along the aryepiglottic folds and arytenoids. The α-gustducin-containing taste buds seem to be located where they may come in contact with food traveling along the lateral food channels in the rat. The taste buds in the rat larynx may work not only as chemical sensors that initiate the reflex reaction but probably participate in gustatory reception. Less

应用DiI逆行标记向喉内肌投射的疑核（NA）神经元，切断其中一条喉返神经，显微分离荧光亮的单个神经元，进行单细胞逆转录（RT）-PCR。免疫组化显示，切断后1、7、10和14 d，NA区许多DiI标记的运动神经元对磷酸化Stat 3呈阳性反应。将30 μ m厚的脑干冰冻切片固定在膜载玻片上，在荧光显微镜下显微解剖NA中DiI标记的单个神经元。通过对单个运动神经元进行实时定量RT-PCR检测，GAP-43和nNOS基因表达水平分别在切断后7天和1天上调。Stat 3、Reg-2和Bcl-2基因表达水平在第7天上调，而B基因表达水平在第7天上调， ...更多信息与对照水平相比，ax在轴突切断后1天和7天下调。应用单细胞RT-PCR结合神经示踪剂DiI标记的新方法，我们可以观察到Stat 3参与轴突切断后向喉肌投射的神经元的部分分子级联反应。香草酸受体在大鼠喉神经支配中的分布我们研究了香草酸受体在大鼠喉神经支配中的分布。VRL-1阳性纤维在喉上皮和固有层中检测到。VR 1阳性神经纤维可见于固有层，但在粘膜上皮中未见。VR 1和VRL-1阳性细胞分布在喉内神经节中，辣椒素受体与VIP、nNOS和cChAT共定位。这些结果表明，这些辣椒素受体参与了喉的副交感神经支配和伤害性感受。α-gustducin在大鼠喉味蕾中的表达我们研究了α-gustducin在大鼠喉味蕾中的表达和分布。α-gustducin阳性味蕾在会厌喉面的尾侧基部、杓会厌襞和杓状软骨沿着分布最密集。含α-味蛋白的味蕾似乎位于它们可能与沿大鼠侧向食物通道沿着行进的食物接触的位置。大鼠喉中的味蕾可能不仅作为化学传感器启动反射反应，但可能参与味觉接收。少