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

先天性巨结肠外源性神经纤维肥大机制的研究：以乙酰胆碱酯酶(AChE)为代表的神经纤维在先天性巨结肠远端无神经节细胞中含量丰富。他的研究目的就是为了阐明这一机制。使用无神经节段大鼠和先天性巨结肠患儿的无神经节段结肠(在手术中提供)。首先，用常规神经标记物--抗PGP9.5抗体对冰冻切片和整装标本的神经纤维分布进行了研究。与对照组相比，无神经节段结肠环肌层神经纤维未见明显增生。TH、NPY(传出交感神经)和Vacht(传出副交感神经)作为盆丛神经纤维的标记物，在环肌层也有同样的表达。P物质和降钙素基因相关肽作为…神经起源的标志物环肌层中肠系膜(传入副交感神经)较少。在粘膜下层，这些神经纤维在血管周围被大量识别。这些结果表明，在无神经节段的结肠内存在两种外源性神经纤维。来自盆神经丛的神经纤维在远端无神经节段肠粘膜下层丰富，而来自肠系膜的神经纤维较少。因此，增多的Ach-E阳性神经主要通过盆丛横穿直肠壁，并出现在无神经节段肠粘膜下层。然后，我们研究了外源性神经在结肠无神经节段的功能方面，使用参与囊泡停靠和融合的蛋白质作为功能标记物。以抗突触素单抗作为囊泡可溶性NSF(N-rthylMaleimide-Sensitive Fusion Protein)结合蛋白受体(SNARE)的标志物，以突触体相关蛋白(SNAP-25)作为靶膜SNAP-25的标记。无神经节段和对照段的粘膜下层均出现陷阱。无神经节段环肌层的神经纤维表现出与囊泡相关的突触小泡吐出功能受损，而不是靶膜。未来的检查内容：有必要研究神经传递受体，以阐明这些神经纤维未来对肠道的作用。较少

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