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 I V 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.
