Receptors for neurotransmitters in Auerbach's plexus of Hirschsprung's disease ; A comparison with animal models

先天性巨结肠症奥尔巴赫丛中的神经递质受体；

基本信息

批准号：
09671237
负责人：
KANEMATSU Takashi
金额：
$ 1.22万
依托单位：
NAGASAKI UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1998
项目状态：
已结题

项目摘要

We examined the changes in receptors of neurotransmitters such as endothelin (EL) and substance P in the intestinal wall related to the dysfunction of large-intestinal motility in Hirselisprung disease and its animal model of congenital aganglionosis (AR) rat, the rat with a mutant endothelin ET_B receptor. We used our newly-developed method for analyzing the receptor dynamics ; the quantitative receptor-imaging system with three experimental techniques, 1) in vitro radioligand-binding technique for functional sites of receptors, 2) quantilative radioimnunohistochernical technique for sites of receptor proteins maturation, and 3) in situ hybridization technique for production sites of receptors with 35S-cRNA probes. We found the existence of ET_B receptors in the main principal cell (cholinergic neuronal cell body) in Auerbach's plexus, and small enteroglia in the colon of patients and AR rats. With the use of our methods, we confirmed the finding that ^1^2^5IRL162O, a selective radioligand for the ET_B receptor, merely bound to the section of colon with Aucrbach's plexus isolated from a patient with Hirschsprung's disease. In this case of patient, we failed to detect a compensatory increase of substance P receptor, using our method. Interestingly, in another patient with Hirschsprung's disease no abnormal binding of ^1^2^5I-1RL1620 was observed, however, a very lower amount of the expression of substance P receptor in Anerbach's plexus. Thus, we confirmed the pathopliysiological significance of the ETB receptor in the onset of Hirschsprung disease, and the possible another gene-defect in this disease. A factor regulating substance P receptor function may shed some light on the discovery of the heterogeneity of Hirschsprung's disease.

我们研究了先天性无神经节细胞病（AR）大鼠模型和先天性无神经节细胞病（AR）大鼠模型中与大肠蠕动功能障碍相关的肠壁内皮素（EL）和P物质等神经递质受体的变化。内皮素ET_B受体。我们使用我们新开发的方法来分析受体动力学；定量受体成像系统采用三种实验技术，1）受体功能位点的体外放射性配体结合技术，2）受体蛋白成熟位点的定量放射免疫组织化学技术，3）受体产生位点的原位杂交技术35S-cRNA 探针。我们在奥尔巴赫神经丛的主要主细胞（胆碱能神经元细胞体）以及患者和 AR 大鼠的结肠小肠神经胶质中发现 ET_B 受体的存在。通过使用我们的方法，我们证实了这样的发现：^1^2^5IRL162O（一种 ET_B 受体的选择性放射性配体）仅与从先天性巨结肠患者分离出的具有奥克巴赫神经丛的结肠部分结合。在该患者的病例中，我们使用我们的方法未能检测到 P 物质受体的代偿性增加。有趣的是，在另一名患有先天性巨结肠症的患者中，没有观察到^1^2^5I-1RL1620的异常结合，然而，Anerbach神经丛中P物质受体的表达量非常低。因此，我们证实了 ETB 受体在先天性巨结肠发病中的病理学意义，以及该疾病中可能存在的另一种基因缺陷。调节 P 物质受体功能的因子可能有助于揭示先天性巨结肠的异质性。