CELL-CELL INTERACTIONS--DYSPLASTIC RETINAL MORPHOGENESIS

本申请中提出的研究将检验这一假设 细胞间的相互作用在正常的视网膜形态发生中起重要作用。这个 主要目标是确定细胞表面糖蛋白和 发育不良视网膜花环形成中的细胞黏附分子。特定的 解决的目标将是：L)以确定是否操纵细胞 表面成分会改变发育中的视网膜的图案形成； 2)确定视网膜花环形成的程度 受操纵细胞表面糖蛋白或黏附的影响 分子。实验旨在从以下方面实现这些目标 几个不同的方面。这项工作将使用三个实验 方法：对处于胚胎的雏鸡进行玻璃体腔注射和 已知可改变特定酶或代谢抑制的新生大鼠 细胞表面黏附分子或糖蛋白作为激发试验 膜微扰；2)器官培养的鸟类和哺乳动物视网膜 更严格地评估和操纵视网膜细胞表面和 在更可控的环境中的粘连现象；3)体外 游离视网膜的再聚集研究将集中在以下过程 玫瑰花环的形成以及修饰对重新聚集的影响 细胞表面。将完成对电池表面的修改 使用特定于糖部分的酶，如内源性尿氨酸酶 神经细胞黏附分子上的聚唾液酸是特异的，或者 作用于其他糖链的其他糖苷酶。细胞表面 糖蛋白成分也将使用代谢抑制剂进行改变。 特异性为N-连接的糖基化或糖蛋白加工位点。 这些操作将通过形态和形态测量学进行评估 视网膜的分析。其他研究将确定细胞的变化 糖蛋白和黏附分子的凝集素组织化学和 细胞黏附分子的免疫组织化学定位。这些 研究对理解正常和异常具有重要意义 以及了解视网膜发育过程中的主要问题之一 实验性视网膜移植的并发症(花环形成)。 它们在一般发育神经生物学和基础神经生物学中有意义 理解细胞-细胞识别。

The studies proposed in this application will test the hypothesis that cell-cell interactions are important in normal retinal morphogenesis. The primary goal is to determine the role of cell surface glycoproteins and cell adhesion molecules in dysplastic retinal rosette formation. Specific objectives addressed will be: l) to determine if manipulation of cell surface components will alter the pattern formation of developing retina; 2) to determine the degree to which retinal rosette formation can be affected by manipulation of cell surface glycoproteins or adhesion molecules. Experiments are designed to approach these objectives from several different aspects. The work will use three experimental approaches: l) intravitreal injections into embryonating chicks and neonatal rats of specific enzymes or metabolic inhibitions known to modify cell surface adhesion molecules or glycoproteins as provocative tests of membrane perturbation; 2) organ cultured avian and mammalian retina will more critically evaluate and manipulate the retinal cell surface and adhesion phenomena in a more controlled environment; 3) in vitro reaggregation studies of dissociated retina will focus on the process of rosette formation and how reaggregation can be affected by modification of the cell surface. Modification of the cell surface will be accomplished using enzymes specific for sugar moieties such as endoneuraminidase which is specific for polysialic acid on neural cell adhesion molecules, or other glycosidases acting at other sugar sties. The cell surface glycoprotein components will also be altered using metabolic inhibitors specific for N-linked glycosylation or sites of glycoprotein processing. These manipulations will be evaluated by morphological and morphometric analysis of the retina. Additional studies will determine changes in cell glycoproteins and adhesion molecules using lectin histochemistry and immunohistochemical localization of cell adhesion molecules. These studies have important implications in understanding normal and aberrant retinal developmental processes as well as understanding one of the major complications of experimental retinal transplantation (rosette formation). They have implications in general developmental neurobiology and the basic understanding of cell-cell recognition.