GLYCOCONJUGATES IN SYMPATHO-ADRENAL DEVELOPMENT

糖复合物在交感肾上腺发育中的作用

• 批准号: 3410842
• 负责人: GERALD A SCHWARTING
• 金额: $ 24.48万
• 依托单位: EUNICE KENNEDY SHRIVER CTR MTL RETARDATN
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-09-25 至 1991-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3410842
• 关键词: adenylate cyclase; adrenal medulla; cell cell interaction; cell differentiation; cell growth regulation; chromaffin cells; cyclic AMP; developmental neurobiology; glucocorticoids; glycolipids; glycoproteins; immunocytochemistry; laboratory mouse; laboratory rat; monoclonal antibody; neural fasciculation; neurons; neurotrophic factors; pheochromocytoma; sympathetic ganglion; tissue /cell culture

The objectives of this proposal are to elucidate the regulation and function of cell surface glycoconjugates in the development of cells of sympatho-adrenal lineage, and to identify specific glycoconjugates as stage- and lineage-specific markers for populations of neurons, chromaffin cells of SIF cells. PC12 pheochromocytoma cells, which exhibit characteristics between chromaffin cells and neurons and can be induced to differentiate in a neuronal direction, will also be utilized as a model for sympatho-adrenal differentiation. These cells acquire further neuronal characteristics after sequential treatment with nerve growth factor (NGF) and activators of adenylate cyclase. PC12 cells express many complex fucose and sialic acid containing glycolipids. In addition, monoclonal antibodies reacting with PC12 cell glycoconjugates have been shown to define developmentally regulated nervous system antigens. In the proposed studies, the interactions of NGF, cAMP and glucocorticoids in regulating PC12 cell glycoconjugate expression will be studied. Expression of these glycoconjugates during normal development will then be established by immunocytochemistry using anti-carbohydrate antibodies. Those glycocojugates which are found to be developmentally regulated, will be studied in primary cultures of adrenal medulla and sympathetic ganglia to determine their sensitivity to regulation by NGF, cyclic AMP and glucocorticoids. Finally, the roles of specific glycoconjugates in neurite outgrowth, cell-cell, and cell-substratum interactions will be assessed. This will be done using two approaches. The expression of specific glycocojugates on subsets of primary cells will be correlated with other phenotypic characteristics, such as the ability to grow neurites, or with commitment to specific differentiation pathways. Secondly, antibodies will be added directly to PC12 and primary cell cultures in attempts to inhibit or retard the expression of those characteristics. These studies provide insight into mechanisms by which physiological messengers such as growth factors and hormones might promote or limit the expression of phenotypic traits associated with sympatho-adrenal derivatives in vivo. In addition, these studies are likely to generate a series of stage- and lineage- specific markers which may be useful for future embryological studies and for characterizing the behavior of a variety of neural and endocrine diseases.

该提案的目的是阐明监管和 细胞表面糖复合物在细胞发育中的功能 交感肾上腺谱系的细胞，并识别特定的 糖缀合物作为阶段和谱系特异性标记 神经元群、SIF 细胞的嗜铬细胞。 电脑12 嗜铬细胞瘤细胞，表现出以下特征： 嗜铬细胞和神经元并可被诱导分化 在神经元方向，也将被用作模型 交感-肾上腺分化。 这些细胞进一步获得 神经序贯治疗后的神经元特征 生长因子（NGF）和腺苷酸环化酶激活剂。 电脑12 细胞表达许多复杂的岩藻糖和唾液酸 糖脂。 此外，与PC12反应的单克隆抗体 细胞糖复合物已被证明可以定义发育 调节神经系统抗原。 在拟议的研究中， NGF、cAMP 和糖皮质激素在调节 PC12 中的相互作用 将研究细胞糖缀合物的表达。 表达 这些糖复合物在正常发育过程中将被 使用抗碳水化合物通过免疫细胞化学建立 抗体。 那些被发现的糖复合物 发育调节，将在原代文化中进行研究 肾上腺髓质和交感神经节来确定它们的 对 NGF、环磷酸腺苷和糖皮质激素调节的敏感性。 最后，特定糖复合物在神经突中的作用 生长、细胞与细胞以及细胞与基质的相互作用将 评估。 这将使用两种方法来完成。 表达式 原代细胞亚群上的特定糖缀合物将是 与其他表型特征相关，例如 生长神经突的能力，或致力于特定的 分化途径。 其次，添加抗体 直接作用于 PC12 和原代细胞培养物，试图抑制 或延迟这些特征的表达。 这些研究提供了对机制的深入了解 生长因子和激素等生理信使可能 促进或限制相关表型性状的表达 与体内交感肾上腺衍生物。 此外，这些 研究可能会产生一系列阶段和谱系 可能对未来胚胎学有用的特定标记 研究和表征各种神经的行为 和内分泌疾病。