SURFACE ANTIGENS OF ORAL BACTEROIDS SPECIES

口腔拟杆菌种的表面抗原

基本信息

批准号：
2128775
负责人：
Ann Progulske-Fox
金额：
$ 7.03万
依托单位：
UNIVERSITY OF FLORIDA
依托单位国家：
美国
项目类别：
财政年份：
1992
资助国家：
美国
起止时间：
1992-04-01 至 1996-03-31
项目状态：
已结题

项目摘要

The present application includes the further development of the genetic system and the generation of site-directed mutants in each of the hemagglutinin (HA) genes. In addition, the application includes approaches to determine if either HA also has protease activity (as circumstantial evidence indicates), to identify functional domains of the HA molecules, and to determine the biological/adhesive function of the HAs using the isogenic mutants in comparison with the wild type parent in in vitro adherence assays. The ultimate goal is to test the virulence of the mutants in animal models. In addition, the erythrocyte receptor molecule will be identified for each HA, employing a variety of biochemical approaches. GRANT=R29DE10150 The long term objectives of these studies are to understand the molecular basis for interactions between epithelial- and mesenchymal-derived tissues that direct tissue morphogenesis, and cell differentiation of mammalian submandibular glands. The hypothesis for this proposal is that epidermal growth factor (EGF) gene products are synthesized by mesenchymal-derived cells in a time- and space-restricted manner and participate in regulating submandibular gland epithelial morphogenesis and differentiation. These EGF gene products interact with receptors on adjacent epithelial cell surfaces and thereby activate signal transduction pathways to convey instructive information from mesenchyme to salivary epithelium. The information is imparted to the epithelial cell nucleus and genome ultimately inducing the expression of a finite family of genes. The epithelial gene products include proteins that i) regulate cell cycle progression; ii) participate in the assembly and degradation of the basement membrane; and iii) induce the expression of cell-specific gene products. To test this hypothesis, the following studies will be performed: i) the temporal and spatial expression of EGF gene products by mesenchymal-derived cells in developing mouse submandibular glands will be studied by high resolution in situ hybridization; ii) simultaneous in situ hybridization and immunochemistry will be used to correlate localized synthesis of EGF gene products with EGF-receptors in the developing mouse submandibular gland; iii) the localized loss or accumulation of specific basement membrane components will be examined in relationship to localized EGF transcription by combined in situ hybridization and immunohistochemistry, and this relationship will be perturbed by the addition of either tyrphostins or antisense EGF oligonucleotides to organ cultures; iv) EGF signal transduction will be perturbed in cultures of developing submandibular glands and the alteration of localized expression of a putative cell cycle control gene, p58, will be determined by in situ hybridization; and v) modification of mitotic indices, tissue morphogenesis and cell differentiation will be determined by mitotic labeling and morphometric analysis in cultured submandibular glands in which EGF signal transduction has been perturbed. The data derived from these studies will contribute to the understanding of molecular mechanisms operant during epithelial-mesenchymal interactions that direct the morphogenesis and differentiation of salivary glands. A knowledge of the molecular mechanisms that direct salivary gland development is critical to understanding the function and behavior of salivary cells as well as the pathogenesis of salivary gland diseases.

本应用包括遗传的进一步发展系统和每个位置指导突变体的产生血凝素（HA）基因。此外，该应用程序包括确定任何一个HA是否也具有蛋白酶活性的方法（AS 间接证据表明），以确定 HA分子，并确定HAS的生物学/粘合功能使用与野生型父母相比，使用等源性突变体体外依从性测定。最终目标是测试动物模型中的突变体。另外，红细胞受体分子将为每个HA确定，采用多种生化方法。格兰特= R29DE10150 这些研究的长期目标是了解分子上皮和间质衍生组织之间相互作用的基础直接组织形态发生和哺乳动物的细胞分化下颌腺。该提议的假设是表皮生长因子（EGF）基因产物由间充质衍生而合成以时间和空间限制的方式进行细胞，并参与调节下颌下皮形态发生和分化。这些 EGF基因产物与相邻上皮细胞上的受体相互作用表面，从而激活信号转导途径以传达从间质到唾液上皮的指导性信息。这信息传授给上皮细胞核和基因组最终诱导有限基因家族的表达。这上皮基因产物包括蛋白质i）调节细胞周期进展； ii）参加大会和退化地下膜； iii）诱导细胞特异性基因的表达产品。为了检验这一假设，以下研究将是进行：i）通过间充质衍生的细胞在发育中的小鼠下颌腺体中将是通过高分辨率原位杂交研究； ii）同时进行原位杂交和免疫化学将用于将局部化发育中的小鼠中EGF基因产物与EGF受体合成下颌腺； iii）特定的局部损失或积累地下室膜组件将在与本地化的关系中进行检查 EGF通过原位杂交和免疫组织化学，这种关系将受到将泰米蛋白或反义EGF寡核苷酸添加到器官文化； iv）EGF信号转导将在培养物中受到干扰开发下颌腺并改变局部表达假定的细胞周期控制基因p58将由原位确定杂交； v）修饰有丝分裂指数，组织形态发生和细胞分化将由有丝分裂确定在培养的下颌腺中的标记和形态分析哪些EGF信号转导已受到干扰。从这些研究将有助于理解分子机制指导上皮 - 间质相互作用期间的操作者唾液腺的形态发生和分化。对直接唾液腺发育的分子机制对于了解唾液细胞的功能和行为以及唾液腺疾病的发病机理。