scholarly journals A role for mouse sperm surface galactosyltransferase in sperm binding to the egg zona pellucida.

1982 ◽  
Vol 95 (2) ◽  
pp. 574-579 ◽  
Author(s):  
B D Shur ◽  
N G Hall
Keyword(s):  
Zona Pellucida ◽  
Milk Protein ◽  
Preceding Paper ◽  
Transferase Activity ◽  
Mouse Sperm ◽  
Sperm Surface ◽  
Galactosyl Transferase ◽  
Sperm Binding ◽  
Beta Galactosidase

Past studies have suggested that mouse sperm surface galactosyltransferase may participate during fertilization by binding N-acetylglucosamine (GlcNAc) residues in the zona pellucida. In this paper, we examined further the role of sperm surface galactosyltransferase in mouse fertilization. Two reagents that specifically perturb sperm surface galactosyltransferase activity both inhibit sperm-zona binding. The presence of the milk protein alpha-lactalbumin specifically modifies the substrate specificity of sperm galactosyltransferase away from GlcNAc and towards glucose and simultaneously inhibits sperm binding to the zona pellucida. Similarly, UDP-dialdehyde inhibits sperm binding to the zona pellucida and sperm surface galactosyl-transferase activity to identical degrees. Of five other sperm enzymes assayed, four are unaffected by UDP-dialdehyde, and one is affected only slightly. Covalent linkage of UDP-dialdehyde to sperm dramatically inhibits binding to eggs, while treatment of eggs with UDP-dialdehyde has no effect on sperm binding. Heat-solubilized or pronase-digested zona pellucida inhibit sperm-zona binding, and they can be glycosylated by sperm with UDP-galactose. Sperm are also able to glycosylate intact zona pellucida with UDP-galactose. Thus, solubilized and intact zona pellucida act as substrates for sperm surface GlcNAc:galactosyltransferases. Finally, pretreatment of eggs with beta-N-acetylglucosaminidase inhibits sperm binding by up to 86%, while under identical conditions, pretreatment with beta-galactosidase increases sperm binding by 55%. These studies, in conjunction with those of the preceding paper dealing with surface galactosyltransferase changes during capacitation, directly suggest that galactosyltransferase is at least one of the components necessary for sperm binding to the zona pellucida.

scholarly journals Receptor function of mouse sperm surface galactosyltransferase during fertilization.

1985 ◽  
Vol 101 (4) ◽  
pp. 1501-1510 ◽  
Author(s):  
L C Lopez ◽  
E M Bayna ◽  
D Litoff ◽  
N L Shaper ◽  
J H Shaper ◽  
...  
Keyword(s):  
Zona Pellucida ◽  
Dorsal Surface ◽  
Mouse Sperm ◽  
Sperm Surface ◽  
Sperm Binding ◽  
Total Inhibition ◽  
Dependent Inhibition ◽  
Dose Dependent Inhibition ◽  
Plasma Membrane Domain ◽  
Dose Dependent

Past studies from this laboratory have suggested that mouse sperm binding to the egg zona pellucida is mediated by a sperm galactosyltransferase (GalTase), which recognizes and binds to terminal N-acetylglucosamine (GlcNAc) residues in the zona pellucida (Shur, B. D., and N. G. Hall, 1982, J. Cell Biol. 95:567-573; 95:574-579). We now present evidence that directly supports this mechanism for gamete binding. GalTase was purified to homogeneity by sequential affinity-chromatography on GlcNAc-agarose and alpha-lactalbumin-agarose columns. The purified enzyme produced a dose-dependent inhibition of sperm binding to the zona pellucida, relative to controls. To inhibit sperm/zona binding, GalTase had to retain its native conformation, since neither heat-inactivated nor Mn++-deficient GalTase inhibited sperm binding. GalTase inhibition of sperm/zona binding was not due to steric blocking of an adjacent sperm receptor on the zona, since GalTase could be released from the zona pellucida by forced galactosylation with UDPGal, and the resulting galactosylated zona was still incapable of binding sperm. In control experiments, when UDPGal was replaced with the inappropriate sugar nucleotide, UDPglucose, sperm binding to the zona pellucida remained normal after the adsorbed GalTase was washed away. The addition of UDPGal produced a dose-dependent inhibition of sperm/zona binding, and also dissociated preformed sperm/zona adhesions by catalyzing the release of the sperm GalTase from its GlcNAc substrate in the zona pellucida. Under identical conditions, UDP-glucose had no effect on sperm binding to the zona pellucida. The ability of UDPGal to dissociate sperm/zona adhesions was both time- and temperature-dependent. UDPGal produced nearly total inhibition of sperm/zona binding when the zonae pellucidae were first galactosylated to reduce the number of GalTase binding sites. Finally, monospecific anti-GalTase IgG and its Fab fragments produced a dose-dependent inhibition of sperm/zona binding and concomitantly blocked sperm GalTase catalytic activity. Preimmune IgG or anti-mouse brain IgG, which also binds to the sperm surface, had no effect. The sperm GalTase was localized by indirect immunofluorescence to a discrete plasma membrane domain on the dorsal surface of the anterior head overlying the intact acrosome. These results, along with earlier studies, show clearly that sperm GalTase serves as a principal gamete receptor during fertilization.

scholarly journals Role of Sperm Surface Arylsulfatase A in Mouse Sperm-Zona Pellucida Binding1

2002 ◽  
Vol 67 (1) ◽  
pp. 212-219 ◽  
Author(s):  
Julierut Tantibhedhyangkul ◽  
Wattana Weerachatyanukul ◽  
Euridice Carmona ◽  
Hongbin Xu ◽  
Araya Anupriwan ◽  
...  
Keyword(s):  
Zona Pellucida ◽  
Arylsulfatase A ◽  
Mouse Sperm ◽  
Sperm Surface

The role of galectin-3 in spermatozoa-zona pellucida binding and its association with fertilization in vitro

2019 ◽  
Vol 25 (8) ◽  
pp. 458-470 ◽  
Author(s):  
Si Mei ◽  
Panyu Chen ◽  
Cheuk-Lun Lee ◽  
Weie Zhao ◽  
Ying Wang ◽  
...  
Keyword(s):  
Seminal Plasma ◽  
Zona Pellucida ◽  
Binding Capacity ◽  
Fertilization Rate ◽  
Clinical Samples ◽  
Fertilization In Vitro ◽  
Human Seminal Plasma ◽  
Sperm Surface ◽  
Galectin 3

AbstractHuman spermatozoa can fertilize an oocyte only after post-testicular maturation and capacitation. These processes involve dynamic modification and reorganization of the sperm plasma membrane, which allow them to bind to the zona pellucida (ZP) of the oocyte. Defective sperm-ZP binding is one of the major causes of male subfertility. Galectin-3 is a secretory lectin in human seminal plasma well known for its action on cell adhesion. The aim of this study was to determine the role of galectin-3 in spermatozoa-ZP interaction and its association with fertilization rate in clinical assisted reproduction. Our studies revealed that the acrosomal region of ejaculated and capacitated spermatozoa possess strong galectin-3 immunoreactivity, which is much stronger than that of epididymal spermatozoa. Expression of galectin-3 can also be detected on seminal plasma-derived extracellular vesicles (EVs) and can be transferred to the sperm surface. Blocking of sperm surface galectin-3 function by antibody or carbohydrate substrate reduced the ZP-binding capacity of spermatozoa. Purified galectin-3 is capable of binding to ZP, indicating that galectin-3 may serve as a cross-linking bridge between ZP glycans and sperm surface glycoproteins. Galectin-3 levels in seminal plasma-derived EVs were positively associated with fertilization rates. These results suggest that galectin-3 in EVs is transferred to the sperm surface during post-testicular maturation and plays a crucial role in spermatozoa-ZP binding after capacitation. Reduced galectin-3 expression in seminal plasma-derived EVs may be a cause behind a low fertilization rate. Further studies with more clinical samples are required to confirm the relationship between galectin-3 levels and IVF outcomes.

scholarly journals Galactosyltransferase activities on mouse sperm bearing multiple tlethal and tviable haplotypes of the T/t-complex

1981 ◽  
Vol 38 (3) ◽  
pp. 225-236 ◽  
Author(s):  
Barry D. Shur
Keyword(s):  
Segregation Distortion ◽  
Enzyme Level ◽  
Transferase Activity ◽  
Wild Type ◽  
Transmission Frequency ◽  
Mouse Sperm ◽  
Sperm Surface ◽  
T Complex ◽  
Specific Increase ◽  
Compound Heterozygotes

SUMMARYSegregation-distorting t-sperm show a specific increase in N-acetylglucosamine: galactosyltransferase activity over wild-type (+/+) due to a deficiency of a wild-type galactosyltransferase inhibitor (Shur & Bennett, 1979). Eight other enzymic activities are indistinguishable between +- and t-sperm suspensions. In this study, three additional points are analysed. First, galactosyltransferases are assayed on sperm homozygous for a semilethal haplotype (tW2/tW2), relative to heterozygous (+ /tw2) and wild-type (+/+) controls. tW2/tW2 assays circumvent the +-sperm inhibition of t-sperm galactosyltransferases that occurs in heterozygous + /t-assays and show that t-sperm are actually four times as active as wild-type. Second, sperm which are compound heterozygotes for two complementing lethal t-haplotypes (tlx/tly), have nearly twice the theoretical enzyme level of tlx/tly sperm. Thus, in either homozygous (tW2/tW2) or double heterozygous (tlx/tly) form t-haplotypes act synergistically on sperm galactosyltransferase activity.Third, and most interesting, sperm bearing either recombinant, viable t-haplotypes (+/tv, tvx/tvy), or one of three dominant T/t-complex mutations, were assayed to determine which portions of the T/t-complex are responsible for elevated galactosyltransferase activity. Results show that sperm bearing recombinant, non-segregation-distorting, viable tv-haplotypes no longer show elevated transferase activity. Therefore, the elevated t1-sperm galactosyltransferase activity strictly correlates with the increased transmission frequency of t1-sperm. These studies strengthen further the hypothesis that sperm surface galactosyltransferases are involved in egg binding during fertilization, and that t1-sperm segregation-distortion results, at least in part, from increased galactosyltransferase activity.

The polysulphate binding domain of human proacrosin/acrosin is involved in both the enzyme activation and spermatozoa-zona pellucida interaction

Zygote ◽  
1998 ◽  
Vol 6 (1) ◽  
pp. 75-83 ◽  
Author(s):  
R. D. Moreno ◽  
M. S. Sepúlveda ◽  
A. de Ioannes ◽  
C. Barros
Keyword(s):  
Zona Pellucida ◽  
Active Site ◽  
Enzyme Activation ◽  
Binding Domain ◽  
Present Evidence ◽  
Sperm Binding ◽  
Mammalian Sperm ◽  
Hydrolysis Of

SummaryMammalian acrosin is a protease present as a zymogen in the acrosome of a non-reacted mammalian sperm, and in vitro is able to carry out limited hydrolysis of homologous and heterologous zonae pellucidae. On the other hand, sulphated polymers and zona pellcida glycoproteins bind to acrosin on a domain different from the active site, named the polysulphate binding domain (PSBD). Thus it is believed that acrosome-reacted spermatozoa bind to glycan chains of the zona pellucida through PSBD participating as secondary binding receptor. The aim of the present work was to study the role of PSBD during both human gamete interaction and acrosin activation. In this work we present evidence that the anti-human acrosin monoclonal antibody C5F10 is directed to an epitope located on or near the PSBD on human proacrosin/acrosin. Moreover, we show that this antibody is able to inhibit both proacrosin activation induced by fucoidan and the sperm binding to the zona pellucida. Our results suggest that the same PSBD is involved in both sperm secondary binding, during zona pellucida penetration, and proacrosin activation.

Interaction of mouse sperm with purified sperm receptors covalently linked to silica beads

1989 ◽  
Vol 92 (4) ◽  
pp. 713-722
Author(s):  
M.H. Vazquez ◽  
D.M. Phillips ◽  
P.M. Wassarman
Keyword(s):  
Zona Pellucida ◽  
Acrosome Reaction ◽  
Solid Phase ◽  
Galactose Oxidase ◽  
Flow Cytofluorometry ◽  
Mouse Sperm ◽  
Sperm Binding ◽  
Silica Beads ◽  
Covalently Linked ◽  
Solid Phase Assay

We describe a solid-phase assay that has permitted further analysis of zona pellucida glycoprotein, ZP3, as sperm receptor and acrosome reaction-inducer during fertilization in mice. The assay employs silica beads that contain epoxy groups to which purified, mouse oocyte ZP3 is covalently linked (ZP3-beads). ZP3-beads were characterized, e.g. by whole-mount autoradiography and flow cytofluorometry, incubated with capacitated mouse sperm under a variety of conditions, and the extent of sperm binding determined by light microscopy. Results of experiments presented suggest the following: (1) sperm bind specifically to ZP3-beads, but not to silica beads either exposed to 2-aminoethanol or derivatized with oocyte ZP2, fetuin or bovine serum albumin. (2) In nearly all cases, only one sperm binds per ZP3-bead and binding occurs via the sperm head. (3) The extent of sperm binding to ZP3-beads is dependent on ZP3 and sperm concentrations, as well as on incubation time and temperature. (4) Sperm binding to ZP3-beads is unaffected by antibodies directed against ZP3, but is inhibited in a reversible manner by treatment of ZP3-beads with galactose oxidase. (5) Only acrosome-intact sperm bind to ZP3-beads but, once bound, sperm can undergo the acrosome reaction, which results in their release from ZP3-beads. (6) Islet-activating protein and 3-quinuclidinyl benzilate, two inhibitors of the zona pellucida-induced acrosome reaction, prevent sperm bound to ZP3-beads from undergoing the acrosome reaction. These results confirm and extend previous studies of sperm-egg interaction in mice, and suggest that the solid-phase assay will be useful for both cellular and biochemical analyses of mammalian fertilization.

scholarly journals ACRBP (Sp32) is involved in priming sperm for the acrosome reaction and the binding of sperm to the zona pellucida in a porcine model

PLoS ONE ◽  
2021 ◽  
Vol 16 (6) ◽  
pp. e0251973
Author(s):  
Yoku Kato ◽  
Satheesh Kumar ◽  
Christian Lessard ◽  
Janice L. Bailey
Keyword(s):  
Endoplasmic Reticulum ◽  
Zona Pellucida ◽  
Acrosome Reaction ◽  
Porcine Model ◽  
Sperm Head ◽  
Mouse Sperm ◽  
Phosphorylated Proteins ◽  
Boar Sperm

In boar sperm, we have previously shown that capacitation is associated with the appearance of the p32 tyrosine phosphoprotein complex. The principal tyrosine phosphoprotein involved in this complex is the acrosin-binding protein (ACRBP), which regulates the autoconversion of proacrosin to intermediate forms of acrosin in both boar and mouse sperm. However, the complete biological role of ACRBP has not yet been elucidated. In this study, we tested the hypothesis that tyrosine phophorylation and the presence of the ACRBP in the sperm head are largely necessary to induce capacitation, the acrosome reaction (AR) and sperm-zona pellucida (ZP) binding, all of which are necessary steps for fertilization. In vitro fertilization (IVF) was performed using matured porcine oocytes and pre-capacitated boar sperm cultured with anti-phosphotyrosine antibodies or antibodies against ACRBP. Anti-ACRBP antibodies reduced capacitation and spontaneous AR (P<0.05). Sperm-ZP binding declined in the presence of anti-phosphotyrosine or anti-ACRBP antibodies. The localisation of anti-ACRBP antibodies on the sperm head, reduced the ability of the sperm to undergo the AR in response to solubilized ZP or by inhibiting the sarco/endoplasmic reticulum Ca2+-ATPase. These results support our hypothesis that tyrosine phosphorylated proteins and ACRBP are present upon the sperm surface in order to participate in sperm-ZP binding, and that ACRBP upon the surface of the sperm head facilitates capacitation and the AR in the porcine.

scholarly journals Role of guinea-pig sperm autoantigens in sperm binding to the zona pellucida and oocyte penetration

Reproduction ◽  
1986 ◽  
Vol 77 (2) ◽  
pp. 347-353 ◽  
Author(s):  
G. M. de Vera ◽  
B. M.-L. Guienne ◽  
M. De Almeida ◽  
G. A. Voisin
Keyword(s):  
Guinea Pig ◽  
Zona Pellucida ◽  
Sperm Binding
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