BIOLOGY OF THE EMBRYONIC CORNEAL EPITHELIUM

描述(调查者摘要)：对基本机制的研究 基底膜参与调节角膜上皮代谢 组件对于理解正常的 角膜上皮细胞、伤口修复过程中的细胞迁移和基底 复发性角膜糜烂和角膜等膜性疾病 营养不良。这笔赠款的长期目标是确定 基底膜和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(粗略 内质网，高尔基体)。我们将检验这一假设 基膜和细胞外基质蛋白决定细胞骨架 通过表面受体进行组织。反过来，细胞骨架也发生了剧烈的变化。 内质网组织或其他转录和 调节蛋白质合成的翻译因子。我已经向你求婚了 将检验这一假设的三个具体目标。1)肌动蛋白 细胞骨架组织其他涉及的细胞骨架蛋白和细胞器 在分泌的蛋白质合成中？2)细胞外基质受到刺激吗？ 胶原合成的增加受转录、翻译和 还是翻译后机制？是协译因素还是粗糙因素？ 通过附着在细胞骨架上而稳定的内质网？3)什么是 所有分泌蛋白质细胞器的空间分布(粗略 内质网、高尔基体和分泌囊)，是它们的 分布依赖于细胞骨架？具体的分布是什么？ 分泌蛋白的信使核糖核酸(胶原蛋白)？是特定的胶原蛋白基因 分布依赖于细胞骨架吗？最近，我开发了一些技术 用共聚焦显微镜观察上皮组织。的发展。 这些技术使我们能够确定空间关系 特定的细胞成分，如细胞骨架蛋白和粗蛋白质 内质网或特定的mRNA。在未来，我希望发展 在活体完整角膜中可视化这些相互作用的技术 上皮组织。综上所述，角膜的细胞生物学模型 细胞外环境与细胞间的上皮细胞相互作用 组织已经发展起来了。新技术将与 经典免疫组织化学、生物化学和分子生物学 细胞骨架元素组织蛋白质分泌的假说 调节分泌物表达的细胞器和翻译辅因子 蛋白质。

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.