Fibroblast Differentiation During Eye Development

眼睛发育过程中成纤维细胞分化

• 批准号: 6875566
• 负责人: GARY W CONRAD
• 金额: $ 36.5万
• 依托单位: KANSAS STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-08-01 至 2009-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6875566
• 关键词: DNA binding protein; Schwann cells; biological signal transduction; biomarker; carbohydrate structure; cell differentiation; chick embryo; clinical research; corneal stroma; developmental neurobiology; extracellular matrix; fibroblasts; genetic regulatory element; glycosylation; histogenesis; human tissue; immunocytochemistry; in situ hybridization; innervation; keratan sulfate; laboratory mouse; laser capture microdissection; mass spectrometry; neural crest; neurotrophic factors; protein biosynthesis; proteoglycan; quail; surface plasmon resonance; transcription factor

DESCRIPTION (provided by applicant): Neural crest-derived, mesenchymal, fibroblastic cell populations differentiate into distinct cell-types in the eye, including corneal nerves, corneal endothelium, stromal keratocytes, non-myelinating and myelinating Schwann cells, connective tissues of limbus, conjunctiva, choroid, sclera, eyelids, iris, lacrimal and other glands, and in birds, scleral ossicles. Each tissue forms a unique extracellular matrix (ECM) that changes with developmental age. Immunocytochemistry, in situ hybridization, mass spectrometry, surface plasmon resonance spectrometry, knock-down and mis-expression analysis, ability to construct chick-quail chimeric embryos, and laser- assisted microdissection (LMD) now allow examination and manipulation of differentiation in small groups of cells in different parts of the eye to better assess signals regulating cell and nerve growth cone migrations and differentiation. Hypothesis: highly charged ECMs unique to each site, bind and release growth-, differentiation-, neurotropic-, and neurotrophic-factors that determine corneal transparency and innervation, and normal functions of other tissues. Aim 1: Determine carbohydrate structures and bound factors in key locations of embryonic eyes. Mass spectrometry will identify and quantitate specific glycosaminoglycans present in ECMs and details of their structures, including structural positions of sulfate groups and identification of sialic acid types. Aim 2: Determine the mechanism of each step in corneal sensory innervation by neural crest-derived growth cones of trigeminal ganglion neurons. Aim 3: Determine the normal lineage sources of non-myelinating Schwann cells of corneal stroma during development; identify marker proteins expressed by these cells and effects on their differentiation from perturbing ECM synthesis. Significance: Extent to which human corneas heal and become re-innervated after LASEK, LASIK, PRK or transplantation is determined by signals residing in participating ECMs, particularly in their posttranslational modifications. Although transplanted corneas generally remain transparent, re-innervation of the graft is very slow (years) and often incomplete, depriving patients of corneal touch sensitivity and blink reflex. Research proposed here will elucidate relationships between proteoglycan forms and distributions, and paths chosen by corneal nerves and non-myelinating Schwann cells during innervation of embryonic corneas. These data will facilitate designing strategies to better reinnervate transplanted or modified adult corneas.

DESCRIPTION (provided by applicant): Neural crest-derived, mesenchymal, fibroblastic cell populations differentiate into distinct cell-types in the eye, including corneal nerves, corneal endothelium, stromal keratocytes, non-myelinating and myelinating Schwann cells, connective tissues of limbus, conjunctiva, choroid, sclera, eyelids, iris, lacrimal和其他腺体，以及在鸟类中，巩膜骨。每个组织形成一个独特的细胞外基质（ECM），随着发育年龄的变化。免疫细胞化学，原位杂交，质谱法，表面等离子体共振光谱法，敲低和错误表达的分析，构造鸡Quail嵌合胚胎的能力以及激光辅助显微解剖（LMD）现在允许在各个部分的细胞中的分化和操纵分化的分化范围和分化的分化范围，以评估各个部分的分化。假设：每个站点独有的高电荷ECM，结合和释放生长，分化 - ，神经旋转和神经营养因素，这些因素决定了角膜透明度和神经支配，以及其他组织的正常功能。目标1：确定胚胎眼睛关键位置的碳水化合物结构和结合因子。质谱法将识别和定量ECM中存在的特定糖胺聚糖及其结构的细节，包括硫酸盐基团的结构位置和唾液酸类型的鉴定。 AIM 2：确定三叉神经节神经元神经衍生的生长锥的角膜感觉神经支配中每个步骤的机制。 AIM 3：确定发育过程中角膜基质的非层状雪旺细胞细胞的正常谱系源；识别这些细胞表达的标记蛋白以及它们与扰动ECM合成的分化的影响。意义：在Lasek，Lasik，Prk或移植后，人角膜愈合并重新发射的程度取决于驻留在参与ECMS中的信号，尤其是在其翻译后修饰中。尽管移植的角膜通常保持透明，但移植物的重新摄入非常缓慢（年），并且通常不完整，剥夺了患者的角膜触摸敏感性和眨眼反射。这里提出的研究将阐明蛋白聚糖形式和分布之间的关系，以及在胚胎角膜神经支配期间，角膜神经和非层状雪旺细胞选择的路径。这些数据将促进设计策略，以更好地重新弥补移植或改良的成人角膜。