Fibroblast Differentiation During Eye Development

神经嵴来源的间充质成纤维细胞群在神经细胞中分化成不同的细胞类型。 眼，包括角膜神经、角膜内皮、基质角膜细胞、非髓鞘形成和髓鞘形成 雪旺细胞、利姆布斯、结膜、脉络膜、巩膜、眼睑、虹膜、泪腺和其他结缔组织 每个组织都形成一种独特的细胞外基质（ECM）， 发育年龄。免疫细胞化学、原位杂交、质谱、表面等离子体 共振光谱法，敲低和错误表达分析，构建鸡-鹌鹑嵌合体的能力 胚胎和激光辅助显微切割（LMD）现在可以检查和操作 在眼睛的不同部分的小细胞群中的分化，以更好地评估调节细胞的信号， 神经生长锥迁移和分化。假设：每个位点独特的高电荷ECM，结合 并释放生长、分化、神经营养和神经营养因子， 透明和神经支配，以及其他组织的正常功能。目的1：测定碳水化合物 胚胎眼睛关键部位的结构和结合因子。质谱法将鉴定和 定量ECM中存在的特定糖胺聚糖及其结构的详细信息，包括结构 硫酸基团的位置和唾液酸类型的鉴定。目标2：确定每种机制 三叉神经节神经元的神经嵴衍生的生长锥在角膜感觉神经支配中的步骤。目的 3：确定在角膜基质中的非髓鞘化雪旺细胞的正常谱系来源。 发育;鉴定这些细胞表达的标记蛋白及其对它们从 干扰ECM合成。意义：人类角膜愈合并重新神经支配的程度 在LASEK、LASIK、LASIK或移植后，由驻留在参与ECM中的信号确定， 特别是在它们的翻译后修饰中。虽然移植的角膜通常 移植物的透明神经再支配非常缓慢（数年），并且通常不完全， 角膜触觉敏感性和眨眼反射。本文提出的研究将阐明 蛋白多糖的形式和分布，以及角膜神经和非髓鞘化雪旺氏神经选择的路径 胚胎角膜神经支配期间的细胞。这些数据将有助于制定战略，以更好地重新 神经支配移植或修饰的成人角膜。

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 re- innervate transplanted or modified adult corneas.