Evidence for the presence of an integrin cell adhesion receptor on the oolemma of unfertilized human oocytes.
The Arg-Gly-Asp peptide (RGD), contained in several extracellular matrix proteins such as fibronectin, laminin, vitronectin, and collagen, is a tripeptide that plays a role as a recognition sequence in many cell-to-cell and cell-to-matrix adhesion mechanisms, through its interaction with several receptors of the integrin family. We previously described the ability of the oolemma of hamster oocytes to bind GRGDTP coupled to the surface of activated immunobeads and demonstrated that RGD-containing oligopeptides inhibit the adhesion of human and hamster spermatozoa to zona-free hamster oocytes and their subsequent penetration. In the present experiments, we show, utilizing immunobeads coated with an RGD-containing peptide (PepTiteTM 2000), that the oolemma of unfertilized human eggs is also able to recognize this adhesion sequence. The binding of PepTiteTM 2000-coated immunobeads to the oolemma was inhibited by the oligopeptide GRGDTP as well as by fibronectin and laminin. When immunobeads were prepared with a PepTiteTM concentration of 10 micrograms/ml, GRGDTP 150 micrograms/ml, laminin 80 micrograms/ml, and fibronectin 60 micrograms/ml inhibited bead rosetting on the egg surface. These data suggest that a specific binding moiety for RGD is present on the human egg surface. The binding of fibronectin to the oolemma was also demonstrated by the rosetting of immunobeads coupled with antifibronectin antibody to human oocytes after their exposure to 1 mg/ml free fibronectin. Such binding of fibronectin to the oolemma could be inhibited by coincubation with a monoclonal antibody directed against the cell adhesion fragment of fibronectin.(ABSTRACT TRUNCATED AT 250 WORDS)
Fusi
F M
FM
III Department of Obstetrics and Gynecology, University of Milano, Italy.
Vignali
M
M
Busacca
M
M
Bronson
R A
RA
D000595:Amino Acid Sequence; D000818:Animals; D001666:Binding Sites, Antibody; D001667:Binding, Competitive; D005260:Female; D005353:Fibronectins / Q000737:chemistry / Q000276:immunology; D006801:Humans; D016023:Integrins / Q000737:chemistry*; D051379:Mice; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000737:chemistry / Q000276:immunology; D009865:Oocytes / Q000737:chemistry*; D011971:Receptors, Immunologic / Q000737:chemistry* / Q000276:immunology; D018000:Receptors, Peptide; D015044:Zona Pellucida / Q000737:chemistry*
D005353:Fibronectins; D016023:Integrins; D009842:Oligopeptides; D011971:Receptors, Immunologic; D018000:Receptors, Peptide; C051382:arginyl-glycyl-aspartic acid directed cell adhesion receptor