GLYCOCONJUGATES IN CYSTIC FIBROSIS: A CELLULAR APPROACH

尽管囊性纤维化(CF)患者的糖蛋白发生了变化 碳水化合物和硫酸盐组成与对照相比， 这些观察结果对所观察到的病理的功能意义 Cf未知。该项目的基本假设解决了这一问题 问题在于，他提出了“人体内分泌物质的凝结 CF患者的外分泌管是由一种异常的 加工或过量生产高阴离子糖偶联物 与阳离子外分泌络合并沉淀的导管上皮 在这个项目中提出的研究将首先确定 阴离子糖偶联物，并在CF成纤维细胞中产生，然后 将这些观察结果与类似的合成和分泌联系起来 外分泌腺导管上皮中的大分子通常 利用利血平化的大鼠感染这种疾病(胰腺) 作为慢性萎缩性胃炎的动物模型。 具体地说，凝集素将用作细胞表面探针，以定量 质膜糖偶联物的碳水化合物组成差异 在正常和CF株的人成纤维细胞中。此外，硫酸盐 正常和CF成纤维细胞的糖蛋白含量也将 分析过了。因为碳水化合物和硫酸盐组成的差异是 检测和定量后，适当的凝集素就可以用作试剂 鉴定成纤维细胞中糖蛋白改变的个别种类 二维凝胶电泳法与碘化凝集素偶联 技巧。然后将进行研究，以分析合成和 硫酸盐化大分子(包括糖共轭化合物)的分泌 CF对外分泌腺导管上皮的影响。在最初的研究中， 上皮中硫酸盐化大分子的合成和分泌 从对照组和利血平治疗组大鼠(A)分离的胰管 建议的CF动物模型)将使用形态学和 生化技术。未来的研究将集中在其他外分泌上 对CF和CF患者分泌液的影响的研究。

Although glycoproteins from Cystic Fibrosis (CF) patients have altered carbohydrate and sulfate composition as compared to controls, the functional significance of these observations to the pathology observed in CF is unknown. The underlying hypothesis of this project addresses this problem by proposing that "the coagulation of secretory material in the exocrine ducts of individuals with CF results from either an abnormal processing or an over-production of highly anionic glycoconjugates from the ductal epithelium which complexe with and precipitate the cationic exocrine glands." The studies proposed in this project will first determine if anionic glyco-conjugates and produced in CF fibroblasts and will then relate these observations to the synthesis and secretion of similar macromolecules in the ductal epithelium of an exocrine gland typically affected in this disease (the pancreas) by ultilizing the reserpinized rat as an animal model of CF. Specifically, lectins will be used as cell surface probes to quantitate differences in the carbohydrate composition of plasmalemmal glycoconjugates in normal and CF strains of human fibroblasts. In addition, the sulfate content of glycoproteins of normal and CF fibroblasts will also be analyzed. As differences in carbohydrate and sulfate composition are detected and quantitated, appropriate lectins can then be used as reagents to identify individual species of glycoproteins altered in CF fibroblasts using 2-D gel electrophoresis couplied with iodinated-lectin overlay techniques. Studies will then be conducted to analyze the synthesis and secretion of sulfated macromolecules (to include glycoconjugates) from the ductal epithelium of exocrine glands affected in CF. In initial studies, the synthesis and secretion of sulfated macromolecules from the epithelium of pancreatic ducts isolated from control and reserpine-treated rats (a proposed animal model of CF) will be analyzed using morphological and biochemical techniques. Future studies will then focus on other exocrine studies affected in CF and on secretory fluids obtained from CF patients.