MUCINS COVERING THE CERVIX AND VAGINA

覆盖子宫颈和阴道的粘蛋白

• 批准号: 2403522
• 负责人: ILENE K. GIPSON
• 金额: $ 29.26万
• 依托单位: SCHEPENS EYE RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-07-01 至 1999-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2403522
• 关键词: age difference; anatomy; cervix; chickens; complementary DNA; enzyme linked immunosorbent assay; epithelium; female reproductive system; gene expression; genetic library; glycosylation; human subject; immunocytochemistry; in situ hybridization; menstrual cycle; mucins; spermicide; topical drug application; vagina

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.

粘蛋白是高度糖基化的糖蛋白，形成一种保护性凝胶 湿的人体上皮。 在女性生殖道中， 子宫颈上皮及其腺体分泌粘蛋白 捍卫子宫和阴道。 使用分子技术 G.I.的特征和呼吸道粘蛋白已产生 尚未应用的有价值的信息和技术技能 研究粘蛋白表达和功能的系统方法 女性生殖道上皮。 该提议汇集了一个 女性生殖医学专家的合作努力和 生物学，眼表面粘蛋白表征和细胞生物学，克隆 胃粘蛋白，粘蛋白特异性抗体的产生及其 在组织中定位。 合作者提出了4个具体目标。 目的 我要确定宫颈和阴道上皮是否表达 表征粘蛋白（粘蛋白1-7）。 AIM II是确定宫颈和 阴道上皮表达独特的粘蛋白。 AIM III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。 AIM IV是根据数据测定 在目标I-III中获得的粘蛋白粘合在阴道上皮并存在 在使用杀精子剂和冲洗之前和之后的阴道中。 技术 使用包括北印迹，原位杂交和 免疫组织化学，使用探针来表征粘蛋白。 表达 使用抗脱糖基化宫颈粘蛋白抗体的克隆技术将 被用来寻找独特的粘蛋白。 比较北印迹， 原位杂交和免疫组织化学将进行确定 解剖区域，年龄和月经周期对粘蛋白的影响 表达。 用于研究局部试剂对粘蛋白的影响 滤纸带衍生表面的阴道，免疫组织化学 将进行粘液和上皮细胞，以及阴道液的ELISA分析 使用来自目标I-III的抗体。 这些研究将提供 有关粘蛋白mRNA和蛋白质表达的基本信息，并将提供 检查局部试剂和病理对影响的工具 宫颈/阴道粘液外套。