title: Production of cartilage link protein by human granulosa-lutein cells.
abstract: Link protein (LP), an extracellular matrix protein in cartilage, stabilizes aggregates of hyaluronic acid (HA) and proteoglycans, including aggrecan and inter-alpha-trypsin inhibitor (ITI). We have shown previously that cartilage LP is present in the maturing rat and mouse ovary. In the present study, we have employed immunohistochemistry to examine the anatomical distribution of cartilage LP in the human ovary. The expression of cartilage LP was selectively detected in the cells within the granulosa compartment of the preovulatory dominant follicle. The HA-positive granulosa-lutein cells were found to be a cartilage LP-positive subpopulation. We subsequently studied the in vitro expression of cartilage LP in cultured human granulosa-lutein cells obtained at oocyte retrieval for in vitro fertilization. Analysis of cultured cells by enzyme-linked immunoaffinity assay, Western blotting and immunofluorescence microscopy revealed that gonadotropin stimulates cartilage LP production. Time-course studies indicated that the cartilage LP production was induced as early as with gonadotropin stimulation for 2 h, and the effect was sustained up to 8 h. Western blot analysis further revealed the presence of the macroaggregates composed of HA, ITI and cartilage LP in the gonadotropin-stimulated granulosa-lutein cell extracts. Collectively, the present results raise the possibility that cartilage LP forms extracellular structures that may have a regulatory function in the developing follicle in the human ovary.
authors.affiliation: Department of Obstetrics and Gynecology, Hamamatsu University School of Medicine, Shizuoka 431-3192, Japan.
authors.forename: G W
authors.initials: GW
authors.lastname: Sun
authors.forename: H
authors.initials: H
authors.lastname: Kobayashi
authors.forename: M
authors.initials: M
authors.lastname: Suzuki
authors.forename: N
authors.initials: N
authors.lastname: Kanayama
authors.forename: T
authors.initials: T
authors.lastname: Terao
mesh_terms: D000328:Adult; D015153:Blotting, Western; D002356:Cartilage / Q000378:metabolism; D002478:Cells, Cultured; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D016326:Extracellular Matrix Proteins / Q000096:biosynthesis*; D005260:Female; D005455:Fluorescent Antibody Technique; D015571:Follicular Fluid / Q000378:metabolism; D006801:Humans; D008184:Luteal Cells / Q000378:metabolism*; D006080:Ovarian Follicle / Q000378:metabolism*; D014176:Protein Biosynthesis; D011509:Proteoglycans; D000099249:Proteoglycan Link Protein
chemical_list: D016326:Extracellular Matrix Proteins; D011509:Proteoglycans; D000099249:Proteoglycan Link Protein