| Project/Area Number |
13470361
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Otorhinolaryngology
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| Research Institution | Kyoto Prefectural University of Medicine |
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Principal Investigator |
HISA Yauso Kyoto Prefectural University of Medicine, Otolaryngology-Head and Neck Surgery, Professor, 医学研究科, 教授 (50181087)
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| Co-Investigator(Kenkyū-buntansha) |
UNO Toshiyuki Kyoto Prefectural University of Medicine, Otolaryngology-Head and Neck Surgery, Associate Professor, 医学研究科, 助教授 (70254349)
SHIMADA Taketoshi Kyoto Prefectural University of Medicine, Otolaryngology-Head and Neck Surgery, Associate Professor, 医学研究科, 助手 (30275226)
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| Project Period (FY) |
2001 – 2003
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| Project Status |
Completed (Fiscal Year 2003)
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| Budget Amount *help |
¥12,600,000 (Direct Cost: ¥12,600,000)
Fiscal Year 2003: ¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 2002: ¥2,900,000 (Direct Cost: ¥2,900,000)
Fiscal Year 2001: ¥6,500,000 (Direct Cost: ¥6,500,000)
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| Keywords | nucleus ambiguus / motonueuron / Stat3 / vanniloid receptors / larynx / gustducin / VRL-1 / 疑核運動神経細胞 / 反回神経 / transsynaptic tracer / 神経再生関連因子 / JAK / STAT系 / GAP-43 / ヘルペスウイルス / アポトーシス関連遺伝子 / motopsin / PRAA12 / nitric oxide synthase / calcitonin gene-related peptide / 画像解析 |
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| Research Abstract |
Molecular changes in neurons of rat nucleus ambiguus after axotomy
Nucleus 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
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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 innervation
We 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 larynx
We 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
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