Localization and Kinetics of Neural Transmitters in the Enteric Nervous System. Studies Using Laser Photoooagulations and Immunochemistry.

肠神经系统中神经递质的定位和动力学。

基本信息

批准号：
62480093
负责人：
KOBAYASHI Shigeru
金额：
$ 4.29万
依托单位：
Yamanashi Medical College
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1987
资助国家：
日本
起止时间：
1987 至 1989
项目状态：
已结题

项目摘要

We investigated nerve regeneration. following lesion forming Argon ion laser irradiation in the guinea-pig small intestine by immunohistochemistry using peroxidase-antiperoxidase complex techniques over a time course of up to 70 days.Round laser-lesions about 0.5mm in diameter were produced and the regenerative features of the two major cell types of the enteric nerve plexuses. i.e. enteric neurons containing neuropeptide by such as methionine-enkephalin-Arg^6-Gly^7-Leu^8 (Enk^<-8>).Peptide-hisbydine-isoleucine (PIII). substance P (SP) and somatostatin (SM) together with tby enteric glial cells containing nerve tissue protein S 100b (S-100b protein by were examined.Immediately after the laser irradiation. the histological structure of the enteric nerve plexuses remained almost intact. though both neuronal and glial cells were coagulated and dead. The immunoreactivities for neuropeptides and S-100b proteins in the neuronal and glial elements respectively were also mostly preserved. By 3 … More hrs after the laser irradiatio, a conspicuous accumulation of immunoreactivities to neuropeptides occurred in the severed nerve stumps. The cellular debris, containing neuropeptides/S-100b protein, were gradually removed from the lesion by 3 days. Since neuropeptides in the neuronal processes are transported from the cell body by the fast anterograde axonal flow, an early accumulation of neuropeptides on the oral edge of the lesion indicates that the neurons project processes in the oro-anal direction, and vice versa.It was deduced that, in the myenteric plexus, both Enk-8 and SP neurons issued both orally and anally directed processes; whereas, PHI and SM neurons sent processes mainly in the oro-anal direct-ion. In the deep muscular plexus (DMP), Enk-8 and SP neurons ran circularly. In the submucous plexus, projections of PHI, SP and SM neurons were directed evenly in all directions.Neuropeptide immunoreactivities of the axotomized nerve cell bodies in the area surrounding the lesion became strikingly stronger 3 to 10 days after laser irradiation. This phenomenon was interpreted as a mode of retrograde degeneration of the enteric neurons. At 15 to 70 days, regenerated nerve plekuses completely spread over the scar tissue of the laser lesion. No nerve cell body, however, existed in the lesioned ganglia. In the regenerated nerve plexus, the glial cells, which proliferated by mitosis, supported and guided the nerve fibers which extended from the survived neuronal cell bodies in the ganglia around the lesion. This suggests that the characteristic neuron/glial cell interactions in the laser lesion might also occur daring the continuous remodeling of the autonomic groundplexus under non-experimental conditions in the enteric nerve plexuses. Less

我们使用过氧化物酶-抗过氧化物酶复合物技术，在长达 70 天的时间过程中，通过免疫组织化学研究了氩离子激光照射豚鼠小肠中形成的损伤后的神经再生。产生了直径约 0.5 毫米的圆形激光损伤，并且肠神经丛的两种主要细胞类型的再生特征，即含有神经肽的肠神经元。蛋氨酸-脑啡肽-Arg^6-Gly^7-Leu^8 (Enk^<-8>)。肽-组氨酸-异亮氨酸(PIII)。P物质(SP)和生长抑素(SM)以及含有肠胶质细胞的激光照射后立即检测神经组织蛋白S 100b（S-100b蛋白）。尽管神经元和神经胶质细胞都凝固并死亡，但在激光照射后 3 小时，神经丛和神经胶质细胞中的神经肽和 S-100b 蛋白的免疫反应性也大部分保留。神经肽免疫反应性的积累发生在切断的神经印记中，含有神经肽/S-100b 蛋白的细胞碎片被逐渐去除。由于神经元突起中的神经肽是通过快速顺行轴突流从细胞体转运的，因此神经肽在病变口腔边缘的早期积累表明神经元在口腔肛门方向上投射突起。据推测，在肌间神经丛中，Enk-8 和 SP 神经元都发出口腔和肛门定向过程，而 PHI 和 SM 神经元主要在肌间神经丛中发出过程；在深肌丛 (DMP) 中，Enk-8 和 SP 神经元循环运行。在粘膜下神经丛中，PHI、SP 和 SM 神经元的投射均匀地定向到各个方向。轴切的神经肽免疫反应性。激光照射后3至10天，病变周围区域的神经细胞体明显增强，这种现象被解释为肠道逆行性变性的一种模式。 15至70天时，再生的神经细胞完全遍布激光损伤的疤痕组织，然而，在再生的神经丛中，通过有丝分裂增殖的神经胶质细胞不存在。支持和引导从病灶周围神经节中幸存的神经元细胞体延伸出来的神经纤维，这表明激光病灶中特征性的神经元/神经胶质细胞相互作用也可能发生。在非实验条件下肠神经丛自主神经丛的持续重塑。