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: 6107225
• 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: 1999
• 资助国家: 美国
• 起止时间: 1999-04-01 至 2000-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6107225
• 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.

我们的研究调查光感受器的细胞和分子生物学 以及它们相关的细胞外基质。 这项研究将开发 参与MBRS学生的生物医学研究技能。 他们将 参与研究的各个方面，包括规划， 执行和评价实验。 他们还将参加会议 提交他们的研究结果，并成为结果出版物的共同作者。 我们 具体目标包括我们研究的新方向以及新的 对持续关注的领域进行调查。 我们将 以章鱼视网膜为模型，具体解决以下问题 系统 1. 感光细胞发育中视网膜特异性mRNA的表达。 利用原位杂交和免疫细胞化学技术， 确定两种特定蛋白质的基因何时首次表达， 正在发育的章鱼视网膜 我们还将使用放射性生物化学和 放射自显影方法来确定功能性视觉色素是否 存在于感光细胞发育的早期。 2. 头足类发色团穿梭机制的激光扫描共聚焦显微镜研究。 视网膜结合蛋白将发色团从髓样体穿梭到 横纹肌 我们将使用激光扫描共聚焦显微镜， 调查这种穿梭机制，并将比较分布 RALBP，视紫红质和视色素在光和暗适应视网膜。 3. 成人和成人视网膜感光细胞间基质的特征 胚胎章鱼视网膜。 生物化学和免疫细胞化学技术 将用于进一步确定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中糖缀合物的存在和身份 从发育的早期阶段到孵化阶段的视网膜。 4. 章鱼类RBP基因的鉴定 分子技术 将被用于鉴定章鱼血清视黄醇结合蛋白的基因 （ORBP）。 这些研究将为继续进行分子生物学研究奠定基础。 章鱼维甲酸转运的研究