THE DEVELOPING AND ADULT VISUAL SYSTEM

视网膜脱离是视力损害的一个严重原因。 占据 了解它的细胞效应可以帮助改善管理， 疾病，并有助于我们的视网膜及其 与色素上皮的关系。 我们最近发现 使用免疫组织化学方法研究长期脱离中特定的细胞和生化变化 动物模型 为了进一步了解这些变化， 结合结构，生物化学和分子技术是一个主要的 这个项目的目标。 具体来说，我们将确定：1）当 中间丝蛋白的上调和 酶和维生素A结合蛋白发生在穆勒细胞（通过使用 免疫细胞化学和免疫印迹分析）; 2）如果mRNA水平， 这些类别中的每一种蛋白质，都随着蛋白质的变化而变化。 蛋白表达（通过北方和狭缝印迹分析;适当的 将通过原位杂交研究mRNA的细胞表达）; 3)光感受器中视蛋白再分布的开始和程度（通过 免疫细胞化学）; 4）如果视蛋白合成和外节更新 继续（通过免疫沉淀和荧光黄带置换 5）视蛋白mRNA水平是否改变以及视蛋白mRNA分布是否改变; 具有重新分布的视蛋白或其形态 严重破坏（通过北方、狭缝印迹和原位杂交 研究）; 6）增殖反应的程度和 所涉及的细胞类型（通过连续递送3 H-胸苷、LM和EM 放射自显影）; 7）如果涉及生长因子（通过 免疫细胞化学）或如果它们可以模拟脱离的效果（通过 将生长因子注射到眼睛中或将其添加到 Muller细胞）; 8）如果蛋白质表达或再分布的变化， 增殖反应被视网膜色素抑制或改变 再附着 对人类视网膜突触组织的更深入了解 通过连续切片电子显微镜研究突触通路， 成年人和发育中的人视网膜的显微镜和高尔基体的分析， 浸渍过的全封。 在后者中，我们将：1）研究染色的 神经元在75个现有的整体安装标准高尔基体描述 技术，以及2）分析进入高尔基体染色细胞的突触输入 通过电子显微镜（通过从供体眼睛获得新的死后组织 并使用各种固定方案来开发更好的结构 视网膜的保存）。

Retinal detachment is a serious cause of visual impairment. Gaining an understanding of its cellular effects can help improve management of the disease and contribute to our basic knowledge of the retina and its relationship to the pigment epithelium. We have recently identified specific cellular and biochemical changes in long-term detachments using an animal model. Gaining a further understanding of these changes using a combination of structural, biochemical, and molecular techniques is a major goal of this project. Specifically we will determine: 1) when the upregulation of intermediate filament proteins and the downregulation of enzymatic and vitamin A binding proteins occurs in Muller cells (by using immunocytochemistry and immunoblot analyses); 2) if the mRNA levels,for one protein in each of these classes, changes in concert with the changes in protein expression (by Northern and slot blot analyses; appropriate cellular expression of the mRNAs will be studied by in situ hybridization); 3) the onset and extent of opsin redistribution in photoreceptors (by immunocytochemistry); 4) if opsin synthesis and outer segment renewal continues (by immunoprecipitation and Lucifer yellow band displacement analyses); 5) if opsin mRNA levels change and if opsin mRNA distribution changes in cells that have redistributed opsin or whose morphology is severely disrupted (by Northern, slot blot and in situ hybridization studies); 6) the extent of the proliferative response and identification of the cell types involved (by continuous delivery of 3H-thymidine, LM and EM autoradiography); 7) if growth factors are involved (by immunocytochemistry) or if they can mimic the effects of detachment (by injection of growth factors into the eye or their addition to cultures of Muller cells); 8) if the changes in protein expression or redistribution or the proliferative response are arrested or modified by retinal reattachment. A greater understanding of the synaptic organization of the human retina will be gained by studying synaptic circuitry by serial-section electron microscopy of adult and developing human retinas and the analysis of Golgi- impregnated whole-mounts. In the latter, we will: 1) study the stained neurons in the 75 existing whole-mounts by standard Golgi descriptive techniques, and 2) analyze the synaptic input into the Golgi stained cells by electron microscopy (by obtaining new postmortem tissue from donor eyes and using a variety of fixation protocols to develop better structural preservation of the retina).