STUDY FOR MECHANISM OF HYPERTROPHIC EXTRINSIC NERVE FIBER IN DISTAL AGANGLIONIC COLON

远端无神经节结肠外源神经纤维肥厚机制的研究

• 批准号: 11671768
• 负责人: WATANABE Yoshio
• 金额: $ 2.24万
• 依托单位: NAGOYA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11671768/
• 关键词: Hirschsprung's disease; extrinsic nerve fibers; pelvic nerve plexus; Acetylcholinesterase; aganglionic colon; SNARE; NO neuron; 神経機能; 腸管神経系

A study of mechanisms for hypertrophic extrinsic nerve fibers in the Hirschsprung's disease colon : Nerve fibers represented with the acetylcholinesterase (AChE) are rich in the distal aganglionic colon of Hirschsprung's disease. The purpose of his study is to clarify this mechanism. The aganglionic colons of both aganglionic rats and Hirschsprung's disease children (provided in an operation) were used. Initially, distribution of the nerve fibers with both frozen section by a microtome and whole-mount preparation was investigated with the anti PGP9.5 antibody, which was a general neural marker. The nerve fibers in the circular muscle layer were not hyperplastic in the aganglionic colon in comparison with control colon. TH, NPY (efferent sympathetic nerve) and VACht (efferent parasympathetic nerve), which were used as marker of the nerve fiber from the pelvic nerve plexus, showed the same in the circular muscle layer. SP and CGRP, which were used as markers for the nerve originated from … More the mesentery (afferent parasympathetic nerve), were poor in the circular muscle layer. In submucosa, these nerve fibers were recognized abundantly in surroundings of the blood vessel. These findings show that there are two kinds of extrinsic nerve fibers in the aganglionic colon. On the other hand, the nerve fibers, which came from a pelvic nerve plexus, were abundant in the submucosa of the distal aganglionic bowel, but the nerve fibers from the mesentery were not. Therefore, increased Ach-E positive nerve mainly traverses in the rectal wall by way of the pelvic *exus, and appears in the submucosal layer of the aganglionic bowel. Then, we studied to clarify the functional aspect of the extrinsic nerve in an aganglionic segment of the colon, using proteins involved in vesicle docking and fusion as a functional marker. Monoclonal antibody for synaptotagmin was prepared as a marker of the soluble NSF (N-rthylmaleimide-sensitive fusion protein) attachment protein receptor (SNARE) in the vesicle and synaptosome-associated protein of Mr 25,000 (SNAP-25) as the SNARE in the target membrane. SNAREs in the submucosa in both the aganglionic segment and control similarly appeared. The nerve fibers in the circular muscle layer of an aganglionic segment showed impaired synaptic vesicle exocytosis related to the vesicle instead of the target membrane.A future examination matter : It is necessary to study receptors of neurotransmission in order to clarify the action to bowel of these nerve fibers in the future. Less

巨结肠病结肠外源性神经纤维肥厚机制的研究：以乙酰胆碱酯酶（AChE）为代表的神经纤维在巨结肠病的远节结结肠中丰富。他的研究目的是阐明这一机制。使用神经节大鼠和先天性巨结肠病儿童的神经节结肠（在手术中提供）。首先，用抗PGP9.5抗体（一种通用的神经标记物）研究了用切片机冷冻切片和全贴装制备的神经纤维的分布。与对照结肠相比，神经节结肠环状肌层神经纤维未见增生。作为盆腔神经丛神经纤维标记物的TH、NPY、VACht在圆形肌层表现出相同的特征。SP和CGRP在圆形肌层中表现较差，在肠系膜（传入副交感神经）中表现较差。在粘膜下层，这些神经纤维在血管周围被充分识别。这些结果表明，在神经节结肠中存在两种外源性神经纤维。另一方面，来自盆腔神经丛的神经纤维在远端神经节肠粘膜下层丰富，而来自肠系膜的神经纤维则不丰富。因此，增加的Ach-E阳性神经主要经盆腔外排穿过直肠壁，出现在神经节肠粘膜下层。然后，我们研究了结肠神经节段外源性神经的功能方面，使用参与囊泡对接和融合的蛋白质作为功能标记。制备synaptotagmin单克隆抗体，作为囊泡内可溶性NSF （n -乙基丙烯酰亚胺敏感融合蛋白）附着蛋白受体（SNARE）和靶膜内Mr 25000突触体相关蛋白（SNAP-25）的标记物。神经节段和对照组的粘膜下层也出现了类似的SNAREs。神经节节段圆形肌层的神经纤维表现出与囊泡有关而非与靶膜有关的突触囊泡胞流出受损。未来的检查事项：有必要研究神经传递受体，以便在未来阐明这些神经纤维对肠道的作用。少

项目成果

Watanabe Y,Ando H,Seo T, et al.: "Two-dimensional Alterations of Myenteric Plexus in Jejunoileal Atresia"Journal of Pediatric Surgery. 36・3. 474-478 (2001)

Watanabe Y、Ando H、Seo T 等：“空肠闭锁肌间神经丛的二维改变”小儿外科杂志 36・3（2001 年）。

Watanabe Y, et al.: "Two-dimensional Alterations of Myenteric Plexus in Jejunoileal Atresia"Journal of Pediatric Surgery. 36・3. 474-478 (2001)

Watanabe Y 等：“空肠闭锁肌间神经丛的二维改变”小儿外科杂志 36・3（2001 年）。

Watanabe Y, et al.: "Morphological Investigation of Enteric Nervous System in Hirschsprung's Disease and Hypoganglionosis Using Whole-mount Colon Preparation"Journal of Pediatric Surgery. 34・3. 445-449 (1999)

Watanabe Y 等人：“利用全结肠准备对先天性巨结肠症和神经节减少症的肠神经系统进行形态学研究”，小儿外科杂志 34・3（1999 年）。

Watanabe Y, et al.: "Morphological Investigation of Enteric Nervous System in Hirschsprung's Disease and Hypoganglionosis Using Whole-mount Colon Preparation"Journal of Pediatric Surgery. 34 3. 445-449 (1999)

Watanabe Y 等人：“使用全结肠制备对先天性巨结肠症和神经节减退症的肠神经系统进行形态学研究”小儿外科杂志。

Watanabe Y, et al.: "Two-dimensional Alterations of Myenteric Plexus in Jejunoileal Atresia"Journal of Pediatric Surgery. 36 3. 474-478 (2001)

Watanabe Y 等人：“空肠闭锁肌间神经丛的二维改变”小儿外科杂志。