title: Incorporation of mouse zona pellucida proteins into the envelope of Xenopus laevis oocytes.
abstract: All vertebrate eggs have extracellular matrices, referred to as the zona pellucida in Mus musculus and the vitelline envelope in Xenopus laevis. The mouse zona, composed of three sulfated glycoproteins (ZP1, ZP2, ZP3), is critical for fertilization and early development, and mice lacking a zona pellucida produce no live offspring. The primary structures of mouse ZP1 (623 amino acids), ZP2 (713 amino acids) and ZP3 (424 amino acids) have been deduced from full-length cDNAs, but posttranslational modifications result in mature zona proteins with molecular masses of 200-180 kDa, 140-120 kDa, and 83 kDa, respectively. The vitelline envelope forms a similar structure around Xenopus eggs and contains three glycoproteins that are structurally related (39-48% amino acid similarity) to the three mouse zona proteins. To investigate whether the structural semblances are sufficient to allow incorporation of the mouse zona proteins into the Xenopus vitelline envelope, capped synthetic mRNAs encoding ZP1, ZP2, and ZP3 proteins were injected into the cytoplasm of stage VI Xenopus oocytes. After 20 h of incubation the oocytes were harvested, and posttranslationally modified zona proteins were detected with monoclonal antibodies specific to mouse ZP1, ZP2, and ZP3. The oocytes were imaged with confocal microscopy to detect individual zona proteins in the extracellular matrix of the oocytes, and this localization was confirmed biochemically. Thus the mouse zona proteins appear to have been sufficiently conserved through 350 million years of evolution to be incorporated into the extracellular envelope surrounding Xenopus eggs.
authors.affiliation: Laboratory of Cellular and Developmental Biology, NIDDK, National Institutes of Health, 6 Center DR MSC 2715, Bethesda, MD 20892-2715, USA. jurrien@helix.nih.gov
authors.forename: S
authors.initials: S
authors.lastname: Doren
authors.forename: N
authors.initials: N
authors.lastname: Landsberger
authors.forename: N
authors.initials: N
authors.lastname: Dwyer
authors.forename: L
authors.initials: L
authors.lastname: Gold
authors.forename: J
authors.initials: J
authors.lastname: Blanchette-Mackie
authors.forename: J
authors.initials: J
authors.lastname: Dean
mesh_terms: D000818:Animals; D004527:Egg Proteins / Q000235:genetics / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D019143:Evolution, Molecular; D005786:Gene Expression Regulation; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D018613:Microscopy, Confocal; D009865:Oocytes / Q000378:metabolism*; D011956:Receptors, Cell Surface; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014817:Vitelline Membrane / Q000737:chemistry / Q000378:metabolism*; D014982:Xenopus laevis; D015044:Zona Pellucida / Q000737:chemistry; D000072216:Zona Pellucida Glycoproteins
chemical_list: D004527:Egg Proteins; D008562:Membrane Glycoproteins; D011956:Receptors, Cell Surface; D011993:Recombinant Fusion Proteins; D000072216:Zona Pellucida Glycoproteins; C000606519:Zp1 protein, mouse; C000606520:Zp2 protein, mouse; C000606521:Zp3 protein, mouse