Characterization of a proteinase that cleaves zona pellucida glycoprotein ZP2 following activation of mouse eggs

Möller, Christer; Wassarman, Paul M.

doi:10.1016/0012-1606(89)90209-1

Cited by 181 publications

(123 citation statements)

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“…We examined whether the oviductal factor responsible for ZP changes resistance to proteolysis and spermoocyte interaction matched any of the previously described factors, such as proteinases (24,25), a peroxidase (26), and a disulfide bond-forming reagent (27) that have been described as factors responsible for cortical granule effects. When pig IVM oocytes were incubated for 30 min in bOF with a mixture of proteinase inhibitors, the ZP did not dissolve even after 206 min in pronase solution, compared with 1 min in control oocytes.…”

Section: Oviduct-specific Glycoprotein Seems To Be Responsible For Thementioning

confidence: 99%

Oviduct-specific glycoprotein and heparin modulate sperm–zona pellucida interaction during fertilization and contribute to the control of polyspermy

Coy

Cánovas

Mondéjar

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

193

177

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Polyspermy is an important anomaly of fertilization in placental mammals, causing premature death of the embryo. It is especially frequent under in vitro conditions, complicating the successful generation of viable embryos. A block to polyspermy develops as a result of changes after sperm entry (i.e., cortical granule exocytosis). However, additional factors may play an important role in regulating polyspermy by acting on gametes before sperm-oocyte interaction. Most studies have used rodents as models, but ungulates may differ in mechanisms preventing polyspermy. We hypothesize that zona pellucida (ZP) changes during transit of the oocyte along the oviductal ampulla modulate the interaction with spermatozoa, contributing to the regulation of polyspermy. We report here that periovulatory oviductal fluid (OF) from sows and heifers increases (both, con-and heterospecifically) ZP resistance to digestion with pronase (a parameter commonly used to measure the block to polyspermy), changing from digestion times of Ϸ1 min (pig) or 2 min (cattle) to 45 min (pig) or several hours (cattle). Exposure of oocytes to OF increases monospermy after in vitro fertilization in both species, and in pigs, sperm-ZP binding decreases. The resistance of OF-exposed oocytes to pronase was abolished by exposure to heparin-depleted medium; in a medium with heparin it was not altered. Proteomic analysis of the content released in the heparin-depleted medium after removal of OFexposed oocytes allowed the isolation and identification of oviduct-specific glycoprotein. Thus, an oviduct-specific glycoproteinheparin protein complex seems to be responsible for ZP changes in the oviduct before fertilization, affecting sperm binding and contributing to the regulation of polyspermy.sperm-oocyte interaction ͉ oviductal fluid ͉ ZP hardening P olyspermy (the penetration of the egg cytoplasm by more than one spermatozoa) is a pathologic condition in placental mammals, usually causing early death of the embryo (1). Although the prevalence of polyspermy under natural conditions is moderate, in in vitro fertilization (IVF) systems polyspermy remains a major obstacle to successful development of viable embryos in different species, including humans (2). Mechanisms underlying the block to polyspermy in mammals have been partially uncovered and characterized, mainly with use of rodents as animal models and usually related to events occurring after sperm entry into the oocyte.The entrance of the spermatozoon into the oocyte's cytoplasm induces the release of cortical granule contents, which modify the vitelline membrane, the zona pellucida (ZP), or both, rendering the oocyte refractory to additional sperm binding and penetration (3) and ending in changes in the mechanical properties and resistance to protease throughout the ZP (4). Yet assuming strong similarities in fertilization mechanisms among rodents and ungulates, observations in ovulated unfertilized porcine and bovine oocytes, showing that ZP resistance to pronase lasts from hours to days (5-8), contras...

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Section: Oviduct-specific Glycoprotein Seems To Be Responsible For Thementioning

confidence: 99%

Oviduct-specific glycoprotein and heparin modulate sperm–zona pellucida interaction during fertilization and contribute to the control of polyspermy

Coy

Cánovas

Mondéjar

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

193

177

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“…The zona block to penetration has been difficult to study separately from mechanisms that affect sperm binding (i.e. if sperm don't bind, they won't penetrate), although historically the block to penetration has been associated with the post-fertilization cleavage of ZP2 (Moller & Wassarman 1989).…”

Section: Post-fertilization Block To Polyspermymentioning

confidence: 99%

Insights into the molecular basis of sperm–egg recognition in mammals

Hoodbhoy¹,

Dean²

2004

Reproduction

126

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The zona pellucida surrounding the egg and pre-implantation embryo is required for in vivo fertility and early development. Explanatory models of sperm -egg recognition need to take into account the ability of sperm to bind to ovulated eggs, but not to two-cell embryos. For the last two decades, investigators have sought to identify an individual protein or carbohydrate side chain as the 'sperm receptor'. However, recent genetic data in mice are more consistent with the three-dimensional structure of the zona pellucida, rather than a single protein (or carbohydrate), determining sperm binding. The mouse and human zonae pellucidae contain three glycoproteins (ZP1, ZP2, ZP3) and, following fertilization, ZP2 is proteolytically cleaved. The replacement of endogenous mouse proteins with human ZP2, ZP3 or both does not alter taxon specificity of sperm binding or prevent fertility. Surprisingly, human ZP2 is not cleaved following fertilization and intact ZP2 correlates with persistent sperm binding to two-cell embryos. Taken together, these data support a model in which the cleavage status of ZP2 modulates the three-dimensional structure of the zona pellucida and determines whether sperm bind (uncleaved) or do not (cleaved).

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“…Several proteinases, probably of CG origin, are released following fertilization and artificial activation and are thought to be involved in ZP modifications that result in blocking polyspermy (Gwatkin et al, 1973;Wolf and Hamada, 1977;Cherr et al, 1988;Moller and Wassarman, 1989;Tawia and Lopata, 1992;Zhang et al, 1992) and "hardening" of the ZP, meaning it acquires increased resistance to proteinases, heat, acid, and reducing agents (reviewed by Gwatkin, 1977;Dunbar, 1983). The molecular basis for hardening is not well understood in mammals, although it may play a role in blocking polyspermy andlor in embryo protection during oviductal transit.…”

Section: Proteinasesmentioning

confidence: 99%

“…ZP, is cleaved from a 120 kD glycoprotein to a 90 kD inactive species (ZP2,) as assessed by 2-D polyacrylamide gel electrophoresis (PAGE) and autoradiography of radiolabeled ZP (Moller and Wassarman, 1989). The activity of this proteinase is insensitive to a large variety of serine-, metallo-, carboxyl-, and sulfahydryl-proteinase inhibitors, although it is inhibited by anti-ZP, antibody (70% inhibition) and by a Fab fragment of this antibody (50% inhibition).…”

Section: Proteinasesmentioning

confidence: 99%

“…The identity of the glycosylated materials is not yet known, but could include glycoproteins such as the CG components discussed below. Proteinases Several proteinases, presumed to be of CG origin, are released from mammalian oocytes following fertilization and artificial activation and have been suggested to establish blocks to polyspermy at the plasma membrane (Wolf and Hamada, 1977) and ZP (Gwatkin et al, 1973;Wolf and Hamada, 1977;Cherr et al, 1988;Moller and Wassarman, 1989;Tawia and Lopata, 1992;Zhang et al, 1992) and to bring about ZP hardening (Moller and Wassarman, 1989;Zhang et al, 1992). In mice, the role of the CG exudate in establishing a plasma membrane block to polyspermy has been controversial.…”

Section: Heparin Binding Placental Protein (Hbpp)mentioning

confidence: 99%

See 1 more Smart Citation

Mammalian cortical granules: Contents, fate, and function

Hoodbhoy

Talbot

1994

Molecular Reproduction Devel

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Recent studies have extended our knowledge regarding the contents of mammalian cortical granules (CG) and their function in postfertilization events. Cytochemical staining has demonstrated the presence of carbohydrates within mammalian CG, and lectin-binding studies have shown that these carbohydrates include a-Dmannose, a-D-GalNAc, and galactose residues in the hamster, a-D-mannose in the mouse and cat, and P-D-Gal(l,3)-D-GalNAc in the pig. Following fertilization and artificial activation, mannosylated material is released from CG and can be found on the oolemma and within the perivitelline space (PVS) of hamster oocytes. Fertilized or artificially activated rabbit, mouse, and human oocytes also release mannosylated, fucosylated and sialylated, and fucosylated material, respectively, which localizes to the oolemma. These glycosylated materials are probably of CG origin, although they have not been directly localized to the CG in rabbit, mice, and humans. The function(s) of the glycosylated material released from mammalian oocytes is not known, although it may participate in blocking polyspermy at the level of the plasma membrane, PVS, and/or zona pellucida (ZP), or it may facilitate preimplantation embryonic development. Proteinases, including tissue plasminogen activator, are also released from mammalian oocytes following fertilization and artificial activation, suggesting that they are of CG origin. These proteinases modify the ZP such that it is no longer receptive to sperm, and some proteinases have been suggested to bring about ZP hardening (an increased resistance to denaturing agents) by an unknown mechanism. Mouse ZP may also be hardened by an ovoperoxidase (cross-links tyrosine residues) cytochemically identified in mouse CG and CG exudate. The phenomena of ZP hardening in mammalian zygotes is not well understood but is likely to function in blocking polyspermic penetration of the ZP and/or in protecting embryos during preirnplantation development. Recently, a 75 kD protein (p75) has been immunocytochemically localized to mouse CG and to the PVS of fertilized oocytes and two-cell embryos. The identity and function of p75 remains to be determined. Heparin binding placental protein may also be a CG component, since it is released from hamster oocytes following fertilization. It has not, however, been directly demonstrated to be a CG component, and its functions in 0 1994 WILEY-LISS, INC.fertilization and/or early embryonic development have yet to be defined. 0 1994 Wlley-Llss, Inc.

show abstract

Characterization of a proteinase that cleaves zona pellucida glycoprotein ZP2 following activation of mouse eggs

Cited by 181 publications

References 43 publications

Oviduct-specific glycoprotein and heparin modulate sperm–zona pellucida interaction during fertilization and contribute to the control of polyspermy

Oviduct-specific glycoprotein and heparin modulate sperm–zona pellucida interaction during fertilization and contribute to the control of polyspermy

Insights into the molecular basis of sperm–egg recognition in mammals

Mammalian cortical granules: Contents, fate, and function

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