MUCINS COVERING THE CERVIX AND VAGINA

粘蛋白是一种高度糖基化的糖蛋白， 尸体表面湿润的上皮细胞 在女性生殖道中， 子宫颈上皮及其腺体分泌粘蛋白 保护子宫和阴道 利用分子技术 G.I.的特征和呼吸道粘蛋白 宝贵的信息和技术技能，尚未应用于 一个系统的方法来研究粘蛋白的表达和功能， 女性生殖道上皮 该提案汇集了一个 女性生殖医学专家的协作努力， 生物学、眼表粘蛋白表征和细胞生物学、克隆 胃粘蛋白，产生粘蛋白特异性抗体及其 在组织中的定位。 合作者提出了四个具体目标。 目的 I是为了确定宫颈和阴道上皮是否表达任何 特征性粘蛋白（粘蛋白1-7）。 目的二是确定宫颈和 阴道上皮表达独特的粘蛋白。 目标三是确定如何 解剖区域、年龄和月经周期影响 鉴定的粘蛋白的表达。 目的四是分析，基于数据 在目的I-III中获得，粘蛋白粘附于阴道上皮并存在于 在使用杀精剂和冲洗之前和之后的阴道中。 技术来 可使用的方法包括北方印迹、原位杂交和 免疫组织化学，使用探针来表征粘蛋白。 表达 使用去糖基化宫颈粘蛋白抗体的克隆技术将 用于寻找独特的粘蛋白。 比较北方印迹法， 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中和上粘蛋白的影响， 阴道，滤纸条表面免疫组织化学 粘液和上皮细胞，并将进行阴道液的ELISA测定 使用源自目的I-III的抗体。 这些研究将提供 粘蛋白mRNA和蛋白质表达的基本信息，并将提供 用于检查局部试剂和病理学对 宫颈/阴道粘液层。

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.