title: The Amount of MUC5B mucin in cervical mucus peaks at midcycle.
abstract: The physical character and amount of mucus secreted by the endocervix changes dramatically during the menstrual cycle to facilitate sperm migration at the time of midcycle ovulation. Mucins are highly glycosylated, high-molecular-weight proteins, which are the major structural components of the protective mucus gel covering all wet-surfaced epithelia, including that of the endocervix. We have previously demonstrated that the endocervical epithelium expresses messenger RNA (mRNA) of three of the large gel-forming mucins, designated MUC5AC, MUC5B, and MUC6, with mRNA of MUC5B predominating. Because mucin protein levels may be regulated posttranscriptionally, measurement of MUC5B protein levels with cycle are needed for correlation to mRNA levels. Measurement of specific mucin gene products within mucus secretions has been limited by availability of specific, well-characterized antibodies and by volume requirements of the isolation protocols for mucins, which include CsCl density centrifugation and fraction isolation. To measure MUC5B protein within the cervical mucus through the hormone cycle, we developed a polyclonal antibody specific to the mucin. The antibody, designated no. 799, is to a synthetic peptide mimicking a 19-amino-acid segment of an intercysteine-rich region within the D4 domain in the 3' region of the MUC5B protein. It recognizes native as well as denatured MUC5B on immunoblot, is preadsorbable with its peptide, and binds to apical secretory vesicles of epithelia expressing MUC5B. We used the MUC5B antibody along with a cervical mucin standard cervical mucin isolate in enzyme-linked immunosorbent assay to determine the relative amount of MUC5B mucin in samples of human cervical mucus taken through the menstrual cycle. We demonstrate a peak of MUC5B mucin in human cervical mucus collected at midcycle, compared with mucus from early or late in the cycle. This peak in MUC5B content coincides with the change in mucus character that occurs at midcycle, suggesting that this large mucin species may be important to sperm transit to the uterus.
authors.affiliation: Schepens Eye Research Institute and Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts 02114, USA. gipson@vision.eri.harvard.edu
authors.forename: I K
authors.initials: IK
authors.lastname: Gipson
authors.forename: R
authors.initials: R
authors.lastname: Moccia
authors.forename: S
authors.initials: S
authors.lastname: Spurr-Michaud
authors.forename: P
authors.initials: P
authors.lastname: Argüeso
authors.forename: A R
authors.initials: AR
authors.lastname: Gargiulo
authors.forename: J A
authors.initials: JA
authors.lastname: Hill
authors.forename: G D
authors.initials: GD
authors.lastname: Offner
authors.forename: H T
authors.initials: HT
authors.lastname: Keutmann
mesh_terms: D000595:Amino Acid Sequence; D000918:Antibody Specificity; D002582:Cervix Mucus / Q000166:cytology / Q000502:physiology*; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000737:chemistry / Q000276:immunology; D005260:Female; D005786:Gene Expression Regulation; D006801:Humans; D007986:Luteinizing Hormone / Q000378:metabolism; D008597:Menstrual Cycle / Q000502:physiology*; D008969:Molecular Sequence Data; D055257:Mucin-5B; D009077:Mucins / Q000032:analysis / Q000097:blood / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis; D012044:Regression Analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012463:Saliva / Q000737:chemistry
chemical_list: D000939:Epitopes; C524578:MUC5B protein, human; D055257:Mucin-5B; D009077:Mucins; D012333:RNA, Messenger; D007986:Luteinizing Hormone