NUCLEAR COMPOSITION AND ORGANIZATION FOR ODONTOGENESIS

牙发生的核组成和组织

• 批准号: 3221707
• 负责人: STEPHEN A SCHWARTZ
• 金额: $ 17.94万
• 依托单位: ROSALIND FRANKLIN UNIV OF MEDICINE & SCI
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-04-01 至 1988-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3221707
• 关键词: basement membrane; cell differentiation; complementary DNA; dental development; embryo /fetus culture; epithelium; gel electrophoresis; gene expression; mesenchyme; messenger RNA; molecular biology; molecular cloning; nucleic acid sequence; oncogenes; organ culture

The long-term objective of the proposed studies is to further extend our understanding of the molecular bases for the initiation and development of the mammalian dental organ. Recently, at least 20 different transformation-specific genetic elements have been found to be normal constituents of most, if not all, eukaryotic genomes. Although many of these genes have been shown to be expressed in a variety of malignant neoplasms, as well as in whole embryos, little if any data exist to identify or clarify the non-neoplastic role(s) of the oncogenes with respect to any single, well-defined developing system. Because the initial histological and proliferative aspects of mammalian toothbud development are reminiscent of neoplastic behavior in the adult, we intend to explore odontogenesis as a likely site for genetically programmed, physiologic "oncogene" expression. We intend to dissect tooth germs from a variety of pre-natal developmental stages of rat embryos, beginning with the earliest evidence for invagination of the primitive oral epithelium. Fist, organs will be reacted for in-situ hybridization against a variety of radiolabeled, oncogene DNA probes representing most of the currently described transforming genes. Second, the extent of physiologic oncogene expression will be correlated with ongoing extracellular matrix composition cell replication, migrations, keratin biosynthesis, and phenotypic differentiation. Third, the expression and appearance of "oncogene"-specific polypeptide gene products will be determined by polyacrylamide gel electrophoretic and blotting methods. Next, the major macromolecular constituents of the matrix, especially laminin, fibronectin, and Type IV collagen will be probed by immunologic detection methods to determine whether "invasive embryologic dental epithelium behaves in a manner comparable to invasive epithelial cells of malignant tumors. Finally, we intend to isolate the DNA from odontogenic epithelium and mesenchymeactively expressing oncogene-specific products. The oncogenic potential of these nucleotide sequences will then be determined following application in an in-vitro oncogene bio-assay method. In this manner, we hope to demonstrate for the first time, unequivocal evidence for programmed, non-neoplastic expression of genetic information currently associated with malignant disease. These studies will greatly enhance our current knowledge regarding the role(s) of the apparently ubiquitous transforming DNA resident within the genomes of all vertebrate cells.

建议研究的长远目标，是进一步扩展我们的 了解启动和发展的分子基础， 哺乳动物的牙齿器官最近，至少有20个不同的 已经发现转化特异性遗传元件是正常的 构成了大多数真核生物基因组的组成部分。 虽然许多 这些基因已被证明在多种恶性肿瘤中表达。 肿瘤，以及在整个胚胎中，几乎没有任何数据存在， 识别或阐明癌基因的非肿瘤作用 任何一个单一的、定义明确的发展系统。 因为初始 哺乳动物牙芽发育组织学和增殖学研究 让人联想到成年人的肿瘤行为，我们打算探索 牙齿发生作为一个可能的网站，遗传程序，生理 “癌基因”表达。 我们打算从各种各样的 大鼠胚胎的产前发育阶段，从最早的 原始口腔上皮内陷的证据。 拳头，器官 将被反应用于原位杂交针对多种 放射性标记的癌基因DNA探针代表了目前大多数 描述了转化基因。 第二，生理性癌基因 表达将与持续的细胞外基质组成相关 细胞复制、迁移、角蛋白生物合成和表型 分化 第三，表达和外观 “癌基因”特异性多肽基因产物将通过 聚丙烯酰胺凝胶电泳和印迹法。 下一篇：The Major 基质的大分子成分，特别是层粘连蛋白，纤连蛋白， 和IV型胶原将通过免疫检测方法探测， 确定“侵入性胚胎牙上皮细胞是否表现为 方式与恶性肿瘤的侵袭性上皮细胞相当。 最后，我们打算从牙源性上皮中分离DNA， 间充质表达癌基因特异性产物。 致癌 这些核苷酸序列的潜力随后将被确定， 在体外癌基因生物测定方法中的应用。 这样，我们 我希望能第一次证明， 目前，遗传信息的程序化非肿瘤表达 与恶性疾病有关。 这些研究将大大提高我们的 目前关于显然无处不在的 转化所有脊椎动物细胞基因组中的DNA。