BIOLOGY OF THE EMBRYONIC CORNEAL EPITHELIUM

胚胎角膜上皮的生物学

• 批准号: 3266242
• 负责人: KATHY Kay SVOBODA
• 金额: $ 12.86万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-07-01 至 1994-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3266242
• 关键词: RNA biosynthesis; actins; basement membrane; cell biology; chick embryo; collagen; confocal scanning microscopy; corneal epithelium; cycloheximide; cytoskeletal proteins; embryo /fetus cell /tissue; endoplasmic reticulum; evaluation /testing; extracellular matrix proteins; immunocytochemistry; in situ hybridization; messenger RNA; organ culture; protein biosynthesis; tissue /cell culture; transcription factor

DESCRIPTION (Investigator's Abstract): The study of the basic mechanisms involved in the regulation of corneal epithelial metabolism by basal lamina components is important to the understanding of normal development of corneal epithelia, cell migration during wound repair, and basement membrane diseases such as recurrent corneal erosions and corneal dystrophies. The long-term objective of this grant is to determine the functional and structural relationships between the basement membrane, corneal epithelial cytoskeleton, and organelle distribution (rough endoplasmic reticulum, Golgi). We will be testing the hypothesis that the basal lamina and extracellular matrix proteins determine cytoskeletal organization via surface receptors. In turn the cytoskeleton changes rough endoplasmic reticulum organization or other transcriptional and translational factors that regulate protein synthesis. I have proposed three specific aims that will test this hypothesis. 1) Does the actin cytoskeleton organize other cytoskeletal proteins and organelles involved in secreted protein synthesis? 2) Is the extracellular matrix-stimulated increase in collagen synthesis regulated by transcriptional, translational, or post-translational mechanism? Are co-translational factors or rough endoplasmic reticulum stabilized by attachment to cytoskeleton? 3) What is the spatial distribution of all secreted protein organelles (rough endoplasmic reticulum, Golgi, and secretory vesicles) and is their distribution cytoskeletal dependent? What is the distribution of specific mRNA for secreted proteins (collage)? Are the specific collagen mRNA distributions cytoskeletal dependent? Recently, I've developed techniques to study epithelial tissues with confocal microscopy. The development of these techniques allows us to determine the spatial relationships between specific cellular components, such as the cytoskeletal proteins and rough endoplasmic reticulum or specific mRNA. In the future, I hope to develop techniques to visualize these interactions in the living intact corneal epithelial tissue. In summary, a cell biological model for corneal epithelia interactions between the extracellular environment and cellular organization has been developed. New techniques will be combined with classical immunohistochemistry, biochemistry, and molecular biology to test the hypothesis that cytoskeletal elements organize protein secretion organelles and translation cofactors to regulate expression of secreted proteins.

描述（研究者摘要）：基本机制的研究 通过基底层参与角膜上皮代谢的调节 组件是重要的理解正常发展的 角膜上皮、创伤修复过程中的细胞迁移和基底 膜疾病，如复发性角膜糜烂和角膜 营养不良 这项补助金的长期目标是确定 基底膜之间的功能和结构关系， 角膜上皮细胞骨架和细胞器分布（粗糙 内质网，高尔基体）。 我们将检验一个假设， 基底层和细胞外基质蛋白决定细胞骨架 组织通过表面受体。 细胞骨架又发生了变化 内质网组织或其他转录和 调节蛋白质合成的翻译因子。 我提出 三个具体目标来检验这个假设。 1)肌动蛋白 细胞骨架组织其他细胞骨架蛋白和细胞器 分泌蛋白质合成中的作用 2)是细胞外基质刺激的 胶原蛋白合成的增加受转录，翻译， 还是翻译后机制 是共翻译因子还是粗糙的 内质网通过附着在细胞骨架上而稳定？ 3)是什么 所有分泌的蛋白质细胞器的空间分布（粗略 内质网，高尔基体和分泌囊泡），是它们的 分布依赖细胞骨架？ 具体的分布情况是什么 分泌蛋白的mRNA（胶原蛋白）？ 是特定的胶原mRNA 分布依赖于细胞骨架？ 最近，我发明了一种技术 用共聚焦显微镜研究上皮组织。 的发展 这些技术使我们能够确定 特定的细胞成分，如细胞骨架蛋白和粗糙蛋白， 内质网或特异性mRNA。 未来，我希望发展 在活体完整角膜中观察这些相互作用的技术 上皮组织 总之，角膜上皮细胞生物学模型 细胞外环境和细胞间的相互作用 组织已经发展起来。 新技术将与 经典的免疫组织化学、生物化学和分子生物学来检测 细胞骨架成分组织蛋白质分泌的假说 细胞器和翻译辅因子来调节分泌的 proteins.