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: 3314769
• 负责人: VIRGINIA M TENNYSON
• 金额: $ 18.86万
• 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/3314769
• 关键词: Schwann cells; antiserum; autoradiography; cell cell interaction; cell migration; collagen; colon disorder; congenital megacolon; degenerative motor system disease; developmental neurobiology; early embryonic stage; electron microscopy; embryo /fetus cell /tissue; extracellular matrix; extracellular matrix proteins; fibronectins; gastrointestinal motility /pressure; gestational age; glia; histochemistry /cytochemistry; immunofluorescence technique; immunoperoxidase; laboratory mouse; microscopy; mucopolysaccharides; neural crest; 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.

致死性斑点突变小鼠（1 S/1 S）在近端出现巨结肠， 无神经节末端肠段。 无神经节细胞组织似乎 从细胞体位于外部的神经元接受轴突的神经支配 内脏 肠内神经元的胞体和突起， 然而，被排除在无神经节区之外。 我们以前使用过 器官型组织培养揭示神经元的分布 前体在正常和1 S/1 S胎鼠肠道。 神经元发育于 培养，如果活的前体存在于肠道中， 解释 这些实验表明，终端2毫米的 任何年龄的突变小鼠的肠道都不会产生神经化的 外植体在文化中，虽然正常小鼠的整个肠道将这样做， 早在E9天。 我们提出，作为一个假设，以解释推导 在1 S/1 S动物中， 在1 S/1 S小鼠中，末端2 mm的肠是异常的，并且不允许 肠神经元前体细胞在该区域的定殖。 这 假设是要测试，如果得到证实，我们将试图确定 缺陷。 以下问题将由拟议的 research. (1)神经元前体不能进入终末2毫米的 1 S/1 S肠还是他们进入这个区域并死亡？ 神经丝 免疫反应性将被用作肠神经元前体标志物 细胞 (2)肠神经胶质细胞的前体不能进入（或存活 在）末端2 mm的1 S/1 S肠？ 胶质细胞的免疫反应性 酸性蛋白将用作神经胶质标记物。 (3)的结构是 末端2 mm的1 S/1 S肠异常？ 是细胞外基质蛋白 1 S/1 S肠末端2 mm处异常？ 免疫细胞 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 旨在利用突变小鼠的缺陷， 组织相互作用在神经系统衍生物发育中的作用 冠。