THE CHEMISTRY OF CELLS AND CELL PORTIONS FROM THE CNS

中枢神经系统细胞和细胞部分的化学性质

• 批准号: 3393242
• 负责人: WILLIAM T NORTON
• 金额: $ 31.3万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 1976
• 资助国家: 美国
• 起止时间: 1976-12-01 至 1995-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3393242
• 关键词: astrocytes; biomarker; cell cycle; cell differentiation; cell growth regulation; cell population study; cell type; experimental allergic encephalomyelitis; experimental brain lesion; gangliosides; glial fibrillary acidic protein; gliosis; growth media; guinea pigs; histochemistry /cytochemistry; immunocytochemistry; laboratory rat; monoclonal antibody; myelin basic proteins; myelination; neurochemistry; neurofilament; oligodendroglia; radiotracer; reporter genes; surface antigens; tissue /cell culture; vimentin; wound healing

The mature central nervous system contains a significant number of immature cells identified by the surface antigen ganglioside GD3. Most of these cells, when isolated and cultured, differentiate to become astrocytes, although some may become oligodendrocytes. The hypotheses to be tested are: that such precursor cells represent the small population of proliferating cells found in adult brain, and that they serve as a source of newly generated astrocytes and oligodendrocytes that appear following traumatic brain lesions and demyelinating events. The specific aims designed to accomplish these goals are: (1) To determine the phenotype of proliferating cells in the normal adult rat CNS. (2) To determine the lineage of proliferating cells in the adult rat CNS following excision lesions and stab wounds. (3) To determine the lineage of proliferating cells in demyelinating and remyelinating lesions in the CNS of guinea pigs with chronic-relapsing experimental autoimmune encephalomyelitis (CREAE). The general experimental design is to identify proliferating cells in tissue sections by either 3H-thymidine incorporation or by the presence of proliferating cell nuclear antigen, a cyclin expressed only in Gl and S phase. In lesioned animals and in CREAE BrdU incorporation will also be used. The phenotype of such cells will then be determined immunohistochemically using stage-specific markers. Besides GD3, such markers will include vimentin, for precursor cells and immature astrocytes, antigen 04, for cells of the oligodendrocyte lineage, GFAP, for differentiated astrocytes, and galactosylceramide and myelin basic protein, for differentiated oligodendrocytes. In addition clonal analysis will be employed with lesioned animals. The BAG retrovirus, carrying the reporter gene, beta-galactosidase, will be injected into the brain at the site of lesion. Animals sacrificed at various times will be processed for immunohistochemistry and X-gal staining to identify the clonal type of cells proliferating in the lesion. Similar procedures will be used on sections of spinal cords of guinea pigs in different stages of CREAE, and in CREAE treated to prevent relapses, to identify the phenotypes of proliferating cells. The results of the proper application of these procedures will provide answers to the following questions. Are proliferating cells in normal and experimental animals solely immature precursors or can mature, differentiated cells also be induced to enter the cell cycle? Do the precursor cells generate astrocytes or oligodendrocytes or both? Are the increased numbers of oligodendrocytes in remyelinating lesions the result of cell proliferation of precursor cells or mature cells? Or do they appear through another mechanism, such as migration of non-proliferating differentiated cells? The health-related aspects of this work stem from the possibility that oligodendrocytes can be replenished in the adult for remyelination in demyelineated lesions such as occur in multiple sclerosis.

成熟的中枢神经系统包含大量 未成熟的细胞由表面抗原神经节苷脂GD3鉴定。 最多 在分离和培养的这些细胞中，分化为 星形胶质细胞，尽管有些可能成为少突胶质细胞。 假设 被测试的是：这样的前体细胞代表小种群 在成人大脑中发现的增殖细胞，它们充当 出现的新生成的星形胶质细胞和少突胶质细胞的来源 经过创伤性脑部病变并脱髓鞘事件。 具体 旨在实现这些目标的目的是：（1）确定 正常成年大鼠中枢神经系统中增殖细胞的表型。 （2）至 确定成年大鼠中枢神经系统中增殖细胞的谱系 切除病变和刺伤。 （3）确定血统 在脱髓鞘和再生病变中的增殖细胞 豚鼠的中枢神经系统带有慢性实验性自身免疫性 脑脊髓炎（CREAE）。 一般的实验设计是 通过任何一个3H-胸腺定鉴定组织切片中的增殖细胞 掺入或通过增殖的细胞核抗原，A Cyclin仅在GL和S相表示。 在病变的动物和 还将使用Creae Brdu融合。 这种细胞的表型 然后将使用特定于阶段的特定于免疫组织化学确定免疫组织化学 标记。 除GD3外，此类标记还包括波形蛋白，用于前体 细胞和未成熟的星形胶质细胞，抗原04，用于细胞 少突胶质细胞谱系，GFAP，用于分化的星形胶质细胞和 半乳糖糖酰胺和髓磷脂碱性蛋白，用于分化 少突胶质细胞。 此外，将使用克隆分析 病变的动物。 袋子逆转录病毒，带有记者基因， β-半乳糖苷酶将被注入 病变。 在不同时间牺牲的动物将被处理 免疫组织化学和X-GAL染色以识别克隆类型 细胞在病变中增殖。 类似的程序将在 在不同阶段的豚鼠的脊髓部分，以及 在经过治疗的Creae中，以防止复发，以识别 增殖细胞。 这些适当应用的结果 程序将为以下问题提供答案。 是 正常动物和实验动物中的增殖细胞仅不成熟 前体或可以成熟的分化细胞也被诱导进入 细胞周期？前体细胞会产生星形胶质细胞或 少突胶质细胞还是两者？少突胶质细胞的数量增加 在恢复病变时，前体细胞增殖的结果 细胞还是成熟的细胞？还是它们是通过另一种机制出现的 作为非增殖分化细胞的迁移？这 这项工作与健康相关的方面源于 可以在成年人中补充少突胶质细胞以在 多发性硬化症中发生的脱髓性病变。