Viral receptors of the visual nervous system

视觉神经系统的病毒受体

• 批准号: 6766527
• 负责人: MARK I GREENE
• 金额: $ 29.32万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-15 至 2008-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6766527
• 关键词: Muller' s cell; Orthoreovirus; affinity chromatography; biological signal transduction; cell growth regulation; developmental neurobiology; embryo /fetus tissue /cell culture; ganglion cell; gene expression; genetically modified animals; glycoproteins; hemagglutinin; laboratory mouse; laboratory rat; nervous system infection; neurogenesis; neuroregulation; newborn animals; pineal body; protein binding; protein protein interaction; protein structure function; surface plasmon resonance; virus infection mechanism; virus receptors; visual pathways

DESCRIPTION (provided by applicant): The objective of this proposal is to extend our understanding of the reovirus type 3 receptor (Reo3R): a complex known to bind to a discrete surface of the reovirus type 3 hemagglutinin (HA3) and to be involved in neural cell differentiation and function. The receptor has been biochemically isolated and found to be composed of two subunits: a 35-65 kD glycosylated protein (CD147) and a 95 kD protein. The 95 kD subunit is a complex of CD147 associated with a multiple membrane-spanning monocarboxylate transporter receptor. Further biological and biochemical studies will be undertaken to examine the use of CD147 as a receptor for reovirus type 3 (T3D) in vivo. Detailed characterization of the biochemistry and features of pathogenesis resulting from binding of T3D to this receptor will be undertaken. Our previous efforts to study receptor signaling focused on the characterization of two structures that bind to this receptor complex. These structures are HA3 and the low affinity monoclonal anti-receptor antibody MAb 87.92.6, particularly one of its complementarity-determining regions from the light chain. We have defined many of the molecular and biological consequences of these interactions. In this proposal, we will focus on the molecular analysis of the reovirus receptor, signal transduction through the receptor, as well as its role in the pathogenesis of neural infection of newborn mice with T3D. We will employ a new set of reagents, including polyclonal and monoclonal antibodies, for this analysis. These reagents bind with high affinity to the Reo3R and block T3D attachment and subsequent infection. In addition, we will continue our efforts to determine the biological role the Reo3R plays in development of neural cells. We will continue to define the receptor's function in the developing optic nervous system and its special role in MuIler and ganglion cells of the retina and in the pineal gland using the high affinity MAb species. We will study two types of transgenic animals: the CD147-/- mouse and a transgenic created by using the interphotoreceptor binding protein (IRBP) promoter coupled with the MAb light chain that recognizes the Reo3R, 87.92.6L. The pineal gland and some retinal cells are genetically ablated in rodents expressing IRBP-87.92.6L. We will study T3D neural pathogenesis in these mutant mice and also define principles of the role of the Reo3R complex in affecting pineal and retinal nerve development.

描述（由申请人提供）：本提案的目的是扩展我们对 3 型呼肠孤病毒受体 (Reo3R) 的理解：一种已知与 3 型呼肠孤病毒血凝素 (HA3) 离散表面结合并参与神经细胞分化和功能的复合物。该受体已被生化分离，发现由两个亚基组成：35-65 kD 糖基化蛋白 (CD147) 和 95 kD 蛋白。 95 kD 亚基是 CD147 与多跨膜单羧酸转运蛋白受体相关的复合物。将进行进一步的生物学和生化研究，以检验 CD147 作为 3 型呼肠孤病毒 (T3D) 体内受体的用途。将进行 T3D 与该受体结合产生的生物化学和发病机制特征的详细表征。 我们之前研究受体信号转导的努力集中于与该受体复合物结合的两种结构的表征。这些结构是HA3和低亲和力单克隆抗受体抗体MAb 87.92.6，特别是其来自轻链的互补决定区之一。我们已经定义了这些相互作用的许多分子和生物学后果。在本提案中，我们将重点关注呼肠孤病毒受体的分子分析、受体的信号转导及其在 T3D 新生小鼠神经感染发病机制中的作用。我们将采用一套新的试剂，包括多克隆和单克隆抗体，用于此分析。这些试剂以高亲和力与 Reo3R 结合，并阻止 T3D 附着和随后的感染。 此外，我们将继续努力确定 Reo3R 在神经细胞发育中所发挥的生物学作用。我们将继续使用高亲和力 MAb 物种来确定受体在发育中的视神经系统中的功能及其在视网膜 MuIler 细胞和神经节细胞以及松果体中的特殊作用。我们将研究两种类型的转基因动物：CD147-/- 小鼠和通过使用光感受器间结合蛋白 (IRBP) 启动子与识别 Reo3R 87.92.6L 的 MAb 轻链偶联而产生的转基因动物。在表达 IRBP-87.92.6L 的啮齿动物中，松果体和一些视网膜细胞被基因消除。我们将研究这些突变小鼠的 T3D 神经发病机制，并确定 Reo3R 复合物在影响松果体和视网膜神经发育中的作用原理。