This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Copyright Permission Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Kobayashi, H. Articles by Terao, T. Search for Related Content PubMed PubMed Citation Articles by Kobayashi, H. Articles by Terao, T.

Identification of Link Protein during Follicle Development and Cumulus Cell Cultures in Rats

Department of Obstetrics and Gynecology, Hamamatsu University School of Medicine, Handacho 3600, Hamamatsu, Shizuoka 431-3192, Japan

Address all correspondence and requests for reprints to: Dr. Hiroshi Kobayashi, Department of Obstetrics and Gynecology, Hamamatsu University School of Medicine, Handacho 3600, Hamamatsu, Shizuoka 431-3192, Japan.

Cumulus oocyte complex (COC) expansion is induced through hyaluronic acid production and accumulation of proteins of the inter--trypsin inhibitor family in the gonadotropin-stimulated cumulus cells. Link protein, a glycoprotein found in cartilage, interacts specifically with hyaluronic acid and stabilizes the binding of proteoglycan monomers to hyaluronic acid to form aggregates. The aim of this study was to investigate the expression of immunoreactive link protein during follicle development in rats and in cumulus cells in culture by immunohistochemistry and Western blot as well as by specific enzyme-linked immunosorbent assay. Immunohistochemical analysis revealed that the extracellular matrix of cumulus cells that were morphologically at a stage of COC expansion were markedly stained for link protein, whereas granulosa cells from immature follicles were not stained. Cumulus cells deposited link protein into the extracellular matrix in an in vitro culture system. The staining intensity was negated by the treatment with hyaluronidase, suggesting that the link protein is bound to hyaluronic acid. We have identified a 42-kDa immunoreactive link protein in rat ovary during the preovulatory period and in COC extracts. Addition of FSH to the medium of cumulus cells in culture supplemented with 10% FBS and oocyte-conditioned medium resulted in an increased rate of link protein synthesis. This work suggests that the cumulus cells synthesize the link protein that may stabilize the binding of inter--trypsin inhibitor or dermatan sulfate proteoglycan to hyaluronic acid to make up hyaluronic acid-rich matrix aggregate.

This article has been cited by other articles:

				S. Kenigsberg, Y. Bentov, V. Chalifa-Caspi, G. Potashnik, R. Ofir, and O. S. Birk Gene expression microarray profiles of cumulus cells in lean and overweight-obese polycystic ovary syndrome patients Mol. Hum. Reprod., February 1, 2009; 15(2): 89 - 103. [Abstract] [Full Text] [PDF]

				D. L. Russell and R. L. Robker Molecular mechanisms of ovulation: co-ordination through the cumulus complex Hum. Reprod. Update, May 1, 2007; 13(3): 289 - 312. [Abstract] [Full Text] [PDF]

				M. Jo and T. E. Curry Jr. Luteinizing Hormone-Induced RUNX1 Regulates the Expression of Genes in Granulosa Cells of Rat Periovulatory Follicles Mol. Endocrinol., September 1, 2006; 20(9): 2156 - 2172. [Abstract] [Full Text] [PDF]

				H. G Clarke, S. A Hope, S. Byers, and R. J Rodgers Formation of ovarian follicular fluid may be due to the osmotic potential of large glycosaminoglycans and proteoglycans Reproduction, July 1, 2006; 132(1): 119 - 131. [Abstract] [Full Text] [PDF]

				M. Schoenfelder and R. Einspanier Expression of Hyaluronan Synthases and Corresponding Hyaluronan Receptors Is Differentially Regulated During Oocyte Maturation in Cattle Biol Reprod, July 1, 2003; 69(1): 269 - 277. [Abstract] [Full Text] [PDF]

				A. P. Spicer, A. Joo, and R. A. Bowling Jr. A Hyaluronan Binding Link Protein Gene Family Whose Members Are Physically Linked Adjacent to Chrondroitin Sulfate Proteoglycan Core Protein Genes: THE MISSING LINKS J. Biol. Chem., May 30, 2003; 278(23): 21083 - 21091. [Abstract] [Full Text] [PDF]

				G. W. Sun, H. Kobayashi, M. Suzuki, N. Kanayama, and T. Terao Follicle-Stimulating Hormone and Insulin-Like Growth Factor I Synergistically Induce Up-Regulation of Cartilage Link Protein (Crtl1) via Activation of Phosphatidylinositol-Dependent Kinase/Akt in Rat Granulosa Cells Endocrinology, March 1, 2003; 144(3): 793 - 801. [Abstract] [Full Text] [PDF]

				L. Zhuo, M. Yoneda, M. Zhao, W. Yingsung, N. Yoshida, Y. Kitagawa, K. Kawamura, T. Suzuki, and K. Kimata Defect in SHAP-Hyaluronan Complex Causes Severe Female Infertility. A STUDY BY INACTIVATION OF THE BIKUNIN GENE IN MICE J. Biol. Chem., March 9, 2001; 276(11): 7693 - 7696. [Abstract] [Full Text] [PDF]

				H. Kobayashi, M. Suzuki, Y. Tanaka, Y. Hirashima, and T. Terao Suppression of Urokinase Expression and Invasiveness by Urinary Trypsin Inhibitor Is Mediated through Inhibition of Protein Kinase C- and MEK/ERK/c-Jun-dependent Signaling Pathways J. Biol. Chem., January 12, 2001; 276(3): 2015 - 2022. [Abstract] [Full Text] [PDF]

				Y. Hirashima, H. Kobayashi, M. Suzuki, Y. Tanaka, N. Kanayama, M. Fujie, T. Nishida, M. Takigawa, and T. Terao Characterization of Binding Properties of Urinary Trypsin Inhibitor to Cell-associated Binding Sites on Human Chondrosarcoma Cell Line HCS-2/8 J. Biol. Chem., April 20, 2001; 276(17): 13650 - 13656. [Abstract] [Full Text] [PDF]