Organization of the Hamster Cumulus Extracellular Matrix: A Hyaluronate-Glycoprotein Gel which Modulates Sperm Access to the Oocyte: Extracellular matrix/Hyaluronate/Oocyte-cumulus complex/Extracellular matrix glycoproteins/Sperm enzymes.

Abstract

The mammalian oocyte-cumulus complex contains an extracellular matrix rich in hyaluronate. Recently, the microstructure of the hamster cumulus extracellular matrix was described (52). In the present work, we investigated the organization of this matrix. We employed freeze-substitution methodologies to investigate ultrastructural effects of various treatments, including sperm enzymes, on the matrix. Protease treatment resulted in disruption with a loss of the fibrillar structures and some expansion; in contrast, hyaluronidase treatment completely solubilized the matrix. EDTA extraction revealed that the fibrils are composed of fine filaments. A discrete region of the matrix immediately surrounding the oocyte, the corona radiata, was resistant to EDTA disruption. We found that hyaluronate is an ubiquitous constituent of the microstructural elements of this extracellular matrix. The matrix exhibits a carbohydrate:protein ratio of approximately 2:1. SDS-PAGE revealed that glycosylated polypeptides are bound to the matrix. The lectins LCA and WGA had differing affinities for these polypeptides, and bound ubiquitously to the intact matrix. The present data suggest that glycoprotein-hyaluronate interaction is critical for maintenance of the cumulus extracellular matrix microstructure and for its physical properties.