MUCINS COVERING THE CERVIX AND VAGINA

粘蛋白是高度糖化的糖蛋白，形成一种保护性凝胶 湿的身体表面上皮细胞。在女性生殖道中， 宫颈及其腺体的上皮细胞分泌粘蛋白，粘蛋白的功能 以保护子宫和阴道。分子技术的应用 GI和呼吸道粘蛋白的特征已经产生 尚未应用的有价值的信息和技术技能 一种系统的研究粘蛋白表达和功能的方法 女性生殖道上皮。这项建议汇集了一个 女性生殖医学专家的合作努力和 生物学，眼表面粘蛋白特性和细胞生物学，克隆 胃粘蛋白、粘蛋白特异性抗体的产生及其 组织中的定位。合作者提出了4个具体目标。目标 我要确定宫颈和阴道上皮是否表达任何 具有粘蛋白特征(粘蛋白1-7)。目标二是确定颈椎和 阴道上皮细胞表达独特的粘蛋白。目标三是确定如何 解剖部位、年龄和月经周期影响模式 已鉴定的粘蛋白的表达。目标四是根据数据进行分析 在AIMS I-III中获得的粘蛋白黏附于阴道上皮并存在 在使用杀精剂和冲洗剂前后的阴道内。技术以达到 包括Northern印迹、原位杂交和 免疫组织化学，用探针表征粘蛋白。表达式 使用脱糖宫颈粘蛋白抗体的克隆技术将 被用来搜索独特的粘蛋白。比较北方印迹， 将进行原位杂交和免疫组织化学以确定 解剖部位、年龄和月经周期对粘蛋白的影响 表情。用于研究局部试剂对粘蛋白的影响 阴道、滤纸条衍生表面的免疫组织化学 粘液和上皮细胞，以及阴道液的酶联免疫吸附试验 使用来自AIMS I-III的抗体。这些研究将提供 有关粘蛋白mRNA和蛋白表达的基本信息，并将提供 检查外用试剂和病理学影响的工具 宫颈/阴道粘液外套。

Mucins are highly glycosylated glycoproteins form a protective gel over the wet surfaced epithelia of the body. In the female reproductive tract, the epithelium of the cervix and its glands secrete mucins which function in defense of the uterus and vagina. Use of molecular techniques for characterization of the G.I. and respiratory tract mucins has yielded valuable information and technical skills which have yet to be applied in a systematic way to the study of mucin expression and function in the female reproductive tract epithelia. This proposal brings together a collaborative effort by experts in female reproductive medicine and biology, ocular surface mucin characterization and cell biology, cloning of gastric mucins, production of mucin-specific antibodies and their localization in tissues. The collaborators propose 4 specific aims. Aim I is to determine if cervical and vaginal epithelia express any of the characterized mucins (mucins 1-7). Aim II is to determine if cervical and vaginal epithelia express unique mucins. Aim III is to determine how anatomical region, age, and menstrual cycle influence pattern of expression of the identified mucins. Aim IV is to assay, based on data obtained in Aims I-III, mucins adherent to vaginal epithelium and present in vagina before and after use of a spermicide and douche. Techniques to be used include northern blotting, in situ hybridization and immunohistochemistry, using probes to characterized mucins. Expression cloning techniques using antibodies to deglycosylated cervical mucins will be employed to search for unique mucins. Comparative northern blotting, in situ hybridization and immunohistochemistry will be done to determine effects of anatomical region, age, and menstrual cycle on mucin expression. For study of effects of topical reagents on mucins in and on the vagina, immunohistochemistry of filter-paper strip-derived surface mucus and epithelial cells, and ELISA assay of vaginal fluids will be done using antibodies derived from Aims I-III. These studies will provide basic information on mucin mRNA and protein expression and will provide tools for examining effects of topical reagents and pathology on cervical/vaginal mucus coat.