CELL AND MOLECULAR BIOLOGY OF PHOTORECEPTORS AND EXTRACELLULAR MATRIX

光感受器和细胞外基质的细胞和分子生物学

• 批准号: 6312764
• 负责人: LAURA J. ROBLES
• 金额: $ 16.39万
• 依托单位: CALIFORNIA STATE UNIV-DOMINGUEZ HILLS
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-04-01 至 2001-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6312764
• 关键词: Cephalopoda; RNA biosynthesis; SDS polyacrylamide gel electrophoresis; alternatives to animals in research; autoradiography; binding proteins; confocal scanning microscopy; electron microscopy; extracellular matrix; gene expression; genetic transcription; immunocytochemistry; in situ hybridization; messenger RNA; molecular biology; nucleic acid probes; rhodopsin; southern blotting; transport proteins; visual photoreceptor; visual pigments

Our research investigates the cell and molecular biology of photoreceptors and their associated extracellular matrix. This research will develop the biomedical research skills of the participating MBRS students. They will be involved in every aspect of the research including the planning, execution, and evaluation of experiments. They will also attend meetings to present their findings and be co-authors on resulting publications. Our specific aims include new direction in our research as well as new investigations of areas that are of continuing interest. We will specifically address the following issues using octopus retina as our model system. 1. The expression of specific retinal mRNAs in photoreceptor development. Using in situ hybridization and immunocytochemical techniques we will determine when the genes for two specific proteins are first expressed in the developing octopus retina. We will also use radiobiochemical and autoradiographic methods to determine if a functional visual pigment is present early in photoreceptor development. 2. LSCM investigation of the chromophore shuttle mechanism in cephalopods. Retinal binding protein shuttles chromophore from the myeloid bodies to the rhabdomes. We will use the Laser Scanning Confocal Microscope to investigate this shuttle mechanism and will compare the distribution of RALBP, rhodopsin and retinochrome in light and dark adapted retinas. 3. Characterization of the interphotoreceptor matrix in adult and embryonic octopus retinas. Biochemical and immunocytochemical techniques will be used to further identify the molecular constituents of the IPM in adult and developing octopus retinas. Lectin binding studies will establish the presence and identity of glycoconjugants in the IPM of retinas from early stages of development through hatching. 4. Identification of an RBP-like gene in octopus. Molecular techniques will be used to identify the gene for octopus serum retinol-binding protein (ORBP). These studies will establish a base for continuing molecular studies on octopus retinoid transport.

我们的研究是研究光感受器的细胞和分子生物学。 及其相关的细胞外基质。这项研究将发展 参与MBR学生的生物医学研究技能。他们会 参与研究的方方面面，包括规划， 实验的执行和评估。他们还将出席会议。 介绍他们的发现，并与人共同撰写由此产生的出版物。我们的 具体目标包括我们研究的新方向以及新的 对持续关注的地区进行调查。我们会 使用章鱼视网膜作为我们的模型，具体解决以下问题 系统。 1.视网膜特异性mRNAs在光感受器发育过程中的表达 利用原位杂交和免疫细胞化学技术，我们将 确定两种特定蛋白质的基因首次在 发育中的章鱼视网膜。我们还将使用放射生物化学和 放射自显影测定功能性视觉色素是否 出现在光感受器发育的早期。 2.头足类发色团穿梭机制的LSCM研究 视网膜结合蛋白将发色团从髓系小体穿梭到 横纹穹顶。我们将使用激光扫描共聚焦显微镜来 研究这种穿梭机制，并将比较 明暗适应视网膜的RALBP、视紫红质和视黄素。 3.成人和成人的光感受器间基质的特征 章鱼胚胎视网膜。生化和免疫细胞化学技术 将被用来进一步鉴定IPM的分子组成 成年和发育中的章鱼视网膜。凝集素结合研究将 确定IPM中糖共轭的存在和身份 视网膜从发育的早期到孵化。 4.章鱼类RBP基因的鉴定。分子技术 将用于鉴定章鱼血清视黄醇结合蛋白的基因 (ORBP)。这些研究将为后续的分子生物学研究奠定基础 章鱼类维甲酸转运的研究。