CONGENITAL MEGACOLON:TISSUE INTERACTIONS IN DEVELOPMENT

先天性巨结肠：发育过程中的组织相互作用

• 批准号: 3314760
• 负责人: VIRGINIA M TENNYSON
• 金额: $ 17.77万
• 依托单位: COLUMBIA UNIV NEW YORK MORNINGSIDE
• 依托单位国家: 美国
• 财政年份: 1983
• 资助国家: 美国
• 起止时间: 1983-12-01 至 1991-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3314760
• 关键词: antiserum; autoradiography; cell cell interaction; cell migration; colon disorder; congenital megacolon; early embryonic stage; electron microscopy; embryo /fetus cell /tissue; extracellular matrix; gastrointestinal motility /pressure; gestational age; glia; histochemistry /cytochemistry; immunofluorescence technique; immunoperoxidase; microscopy; mucopolysaccharides; neurofilament; organ culture; pathologic process; stainings

The lethal spotted mutant mouse (1S/1S) develops megacolon proximal to a segment of aganglionic terminal bowel. The aganglionic tissue appears to receive an innervation of axons from neurons whose cell bodies lie outside the gut. The cell bodies and processes of intrinsic enteric neurons, however, are excluded from the aganglionic region. We have previously used organotypic tissue cultures to reveal the distribution of neuronal precursors in the normal and 1S/1S fetal mouse gut. Neurons develop in culture if viable precursors are present in the gut at the time of explanation. These experiments have revealed that the terminal 2 mm of bowel from mutant mice at any age will never give rise to neuralized explants in culture, although the entire gut of normal mice will do so as early as day E9. we propose, as a hypothesis to account for the derivation of aganglionosis in 1S/1S animals, that the microenvironment of the terminal 2mm of bowel is abnormal in 1S/1S mice and does not permit the colonization of that region with enteric neuronal precursor cells. This hypothesis is to be tested and, if confirmed, we will attempt to identify the defect. The following questions will be answered by the proposed research. (1) Do neuronal precursors fail to enter the terminal 2 mm of 1S/1S bowel or do they enter this region and die? Neurofilament immunoreactivity will be used as a marker for enteric neuronal precursor cells. (2) Do precursors of enteric glial cells fail to enter (or survive in) the terminal 2 mm of 1S/1S bowel? Immunoreactivity of glial fibrillary acid protein will be used as a glial marker. (3) Is the structure of the terminal 2 mm of 1S/1S bowel abnormal? Are extracellular matrix proteins abnormal in the terminal 2 mm of 1S/1S bowel? Immunocytochemical examination of tissues will be done to determine the distribution of fibronectin, laminin, and types I and IV collagen. These experiments are designed to exploit the defect of the mutant mouse to gain insight into the role of tissue interactions in the development of derivatives of the neural crest.

致命的斑点突变小鼠（1S/1S）在靠近 无神经节末端肠段。 无神经节组织似乎 接受来自细胞体位于外部的神经元的轴突神经支配 肠道。 内在肠神经元的细胞体和过程， 然而，被排除在无神经节区域之外。 我们之前用过 器官型组织培养揭示神经元的分布 正常和 1S/1S 胎儿小鼠肠道中的前体。 神经元发育于 培养时肠道中是否存在可行的前体 解释。 这些实验表明，末端2毫米 任何年龄的突变小鼠的肠道都不会产生神经化 培养物中的外植体，尽管正常小鼠的整个肠道都会这样做 早在 E9 天。 我们提出，作为解释推导的假设 1S/1S 动物无神经节细胞病的微环境 1S/1S 小鼠的肠末端 2mm 是异常的，不允许 肠神经元前体细胞在该区域定植。 这 假设需要进行检验，如果得到证实，我们将尝试识别 缺陷。 提案将回答以下问题 研究。 (1)神经元前体是否未能进入神经元末端2mm？ 1S/1S 肠道还是进入该区域并死亡？ 神经丝 免疫反应性将用作肠神经元前体的标记 细胞。 （2）肠胶质细胞的前体细胞是否无法进入（或存活） in) 1S/1S 肠末端 2 毫米？ 胶质原纤维的免疫反应性 酸性蛋白将用作神经胶质标记物。 (3) 结构是 1S/1S肠末端2毫米异常？ 是细胞外基质蛋白 1S/1S肠末端2毫米异常？ 免疫细胞化学 将进行组织检查以确定 纤连蛋白、层粘连蛋白以及 I 型和 IV 型胶原蛋白。 这些实验是 旨在利用突变小鼠的缺陷来深入了解 组织相互作用在神经元衍生物发育中的作用 波峰。