scholarly journals Autoradiographic visualization of the mouse egg's sperm receptor bound to sperm.

1986 ◽  
Vol 102 (4) ◽  
pp. 1363-1371 ◽  
Author(s):  
J D Bleil ◽  
P M Wassarman
Keyword(s):  
Plasma Membrane ◽  
Zona Pellucida ◽  
Present Report ◽  
Fusion Event ◽  
Acrosomal Membrane ◽  
Specific Receptors ◽  
Species Specific ◽  
Mammalian Eggs ◽  
Silver Grains ◽  
Sperm Receptor

The extracellular coat, or zona pellucida, of mammalian eggs contains species-specific receptors to which sperm bind as a prelude to fertilization. In mice, ZP3, one of only three zona pellucida glycoproteins, serves as sperm receptor. Acrosome-intact, but not acrosome-reacted, mouse sperm recognize and interact with specific O-linked oligosaccharides of ZP3 resulting in sperm-egg binding. Binding, in turn, causes sperm to undergo the acrosome reaction; a membrane fusion event that results in loss of plasma membrane at the anterior region of the head and exposure of inner acrosomal membrane with its associated acrosomal contents. Bound, acrosome-reacted sperm are able to penetrate the zona pellucida and fuse with the egg's plasma membrane (fertilization). In the present report, we examined binding of radioiodinated, purified, egg ZP3 to both acrosome intact and acrosome reacted sperm by whole-mount autoradiography. Silver grains due to bound 125I-ZP3 were found localized to the acrosomal cap region of heads of acrosome-reacted sperm. Under the same conditions, 125I-fetuin bound at only bacKground levels to heads of both acrosome-intact and -reacted sperm, and 125I-ZP2, another zona pellucida glycoprotein, bound preferentially to acrosome-reacted sperm. These results provide visual evidence that ZP3 binds preferentially and specifically to heads of acrosome intact sperm; properties expected of the mouse egg's sperm receptor.

Profile of a mammalian sperm receptor

Development ◽  
1990 ◽  
Vol 108 (1) ◽  
pp. 1-17 ◽  
Author(s):  
P.M. Wassarman
Keyword(s):  
Gene Expression ◽  
Zona Pellucida ◽  
Specific Gene ◽  
Mammalian Development ◽  
Unique Nature ◽  
Mammalian Fertilization ◽  
Species Specific ◽  
Available Information ◽  
Mammalian Eggs ◽  
Sperm Receptor

Complementary molecules on the surface of eggs and sperm are responsible for species-specific interactions between gametes during fertilization in both plants and animals. In this essay, several aspects of current research on the mouse egg receptor for sperm, a zona pellucida glycoprotein called ZP3, are addressed. These include the structure, synthesis, and functions of the sperm receptor during oogenesis and fertilization in mice. Several conclusions are drawn from available information. These include (I) ZP3 is a member of a unique class of glycoproteins found exclusively in the extracellular coat (zona pellucida) of mammalian eggs. (II) ZP3 gene expression is an example of oocyte-specific and, therefore, sex-specific gene expression during mammalian development. (III) ZP3 is a structural glycoprotein involved in assembly of the egg extracellular coat during mammalian oogenesis. (IV) ZP3 is a sperm receptor involved in carbohydrate-mediated gamete recognition and adhesion during mammalian fertilization. (V) ZP3 is an inducer of sperm exocytosis (acrosome reaction) during mammalian fertilization. (VI) ZP3 participates in the secondary block to polyspermy following fertilization in mammals. (VII) The extracellular coat of other mammalian eggs contains a glycoprotein that is functionally analogous to mouse ZP3. The unique nature, highly restricted expression, and multiple roles of ZP3 during mammalian development make this glycoprotein a particularly attractive subject for investigation at both the cellular and molecular levels.

Differential binding of gold-labeled zona pellucida glycoproteins mZP2 and mZP3 to mouse sperm membrane compartments

Development ◽  
1991 ◽  
Vol 113 (1) ◽  
pp. 141-149 ◽  
Author(s):  
S. Mortillo ◽  
P.M. Wassarman
Keyword(s):  
Plasma Membrane ◽  
Zona Pellucida ◽  
Transmission Electron ◽  
Acrosomal Membrane ◽  
Membrane Compartments ◽  
Sperm Membrane ◽  
Differential Binding ◽  
Inner Acrosomal Membrane ◽  
Low Levels ◽  
Species Specific

Egg zona pellucida glycoproteins mZP3 and mZP2 serve as primary and secondary sperm receptors, respectively, during initial stages of fertilization in mice [Wassarman (1988) A. Rev. Biochem. 57, 415–442]. These receptors interact with complementary egg-binding proteins (EBPs) located on the sperm surface to support species-specific gamete adhesion. Results of whole-mount autoradiographic experiments suggest that purified egg mZP3 and mZP2 bind preferentially to acrosome-intact (AI) and acrosome-reacted (AR) sperm heads, respectively [Bleil and Wassarman (1986) J. Cell Biol. 102, 1363–1371]. Here, we used purified egg mZP2, egg mZP3 and fetuin, which were coupled directly to colloidal gold (‘gold-probes’), to examine binding of these glycoproteins to membrane compartments of AI and AR sperm by transmission electron microscopy. mZP3 gold-probes were found associated primarily with plasma membrane overlying the acrosomal and post-acrosomal regions of AI sperm heads. They were also found associated with plasma membrane overlying the post-acrosomal region of AR sperm heads. mZP2 gold-probes were found associated primarily with inner acrosomal membrane of AR sperm heads, although some gold was associated with outer acrosomal membrane of AI sperm that had holes in plasma membrane overlying the acrosome. Fetuin gold-probes, used to assess background levels of binding, were bound at relatively low levels to plasma membrane and inner acrosomal membrane of AI and AR sperm, respectively. None of the gold-probes exhibited significant binding to sperm tails, or to red blood cells and residual bodies present in sperm preparations. These results provide further evidence that mZP2 and mZP3 bind preferentially to heads of AR and AI sperm, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Arachidonic acid-induced acrosomal loss in the spermatozoa of a marsupial, the tammar wallaby (Macropus eugenii)

1997 ◽  
Vol 9 (8) ◽  
pp. 803 ◽  
Author(s):  
Yulia Sistina ◽  
John C. Rodger
Keyword(s):  
Arachidonic Acid ◽  
Plasma Membrane ◽  
Membrane Fusion ◽  
Tammar Wallaby ◽  
Multiple Point ◽  
Fusion Event ◽  
Macropus Eugenii ◽  
Kinase C ◽  
Acrosomal Membrane ◽  
Late Stages

Tammar wallaby spermatozoa were induced to undergo acrosomal loss when incubated with arachidonic acid (AA). Ultrastructural examination indicated that the AA-induced acrosomal loss occurred via multiple point fusions between the outer acrosomal membrane and the overlying plasma membrane. This form of acrosomal loss mimicked the physiological acrosome reaction (AR) seen in the sperm of eutherian mammals. The fusion event was limited to the acrosomal region of the plasma membrane and did not proceed past the peri-acrosomal ring. The entire acrosome was lost after AA treatment leaving no evidence of a persistent equatorial segment-like region. Ultrastructural evidence of AR-like membrane fusion was seen immediately on addition of 50 µg mL-1 AA and a large proportion of sperm examined after five min incubation were in the late stages of membrane fusion. Longer-term incubation with AA had deleterious effects on wallaby sperm motility. It remains to be determined whether the AA-induced membrane fusion observed here indicates that AA is involved in the marsupial AR. However, pretreatment of sperm with the protein kinase C (PKC) inhibitor HMG significantly reduced AA-induced acrosomal loss suggesting that AA may have acted via PKC. If this is so, AA is probably physiologically significant and a novel pathway may be operating during AR induction in marsupials.

scholarly journals Mammalian membrane block to polyspermy: new insights into how mammalian eggs prevent fertilisation by multiple sperm

2006 ◽  
Vol 18 (2) ◽  
pp. 53 ◽  
Author(s):  
Allison J. Gardner ◽  
Janice P. Evans
Keyword(s):  
Plasma Membrane ◽  
Zona Pellucida ◽  
Animal Species ◽  
Mammalian Species ◽  
Experimental Studies ◽  
Present Review ◽  
Cellular Signalling ◽  
Classical Studies ◽  
Vitelline Envelope ◽  
Mammalian Eggs

To inhibit fertilisation by more than one sperm (a condition known as polyspermy), eggs have developed preventative mechanisms known as blocks to polyspermy. The block at the level of the egg extracellular coat (the zona pellucida in mammals, the vitelline envelope in non-mammals) has been well characterised in many different animal species and the block at the level of the egg plasma membrane is understood in some non-mammalian species. However, virtually nothing is known about the membrane block to polyspermy in mammalian eggs, despite data dating back 50–90 years that provide evidence for its existence. In the present review, we will discuss the background on blocks to polyspermy used by animal eggs and then focus on the membrane block to polyspermy in mammalian eggs. This will include a summary of classical studies that provide evidence for this block in mammalian eggs, assays used to study the mammalian membrane block and what has been elucidated from recent experimental studies about the cellular signalling events that lead to membrane block establishment and the mechanism of how the membrane block may prevent additional fertilisation.

scholarly journals Update on mammalian sperm capacitation: how much does the horse differ from other species?

Reproduction ◽  
2019 ◽  
Vol 157 (5) ◽  
pp. R181-R197 ◽  
Author(s):  
Bart Leemans ◽  
Tom A E Stout ◽  
Catharina De Schauwer ◽  
Sonia Heras ◽  
Hilde Nelis ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Zona Pellucida ◽  
Mammalian Species ◽  
Cumulus Cells ◽  
Sperm Cells ◽  
Cholesterol Depletion ◽  
Sperm Capacitation ◽  
Sperm Plasma Membrane ◽  
Species Specific

In contrast to various other mammalian species, conventional in vitro fertilization (IVF) with horse gametes is not reliably successful. In particular, stallion spermatozoa fails to penetrate the zona pellucida, most likely due to incomplete activation of stallion spermatozoa (capacitation) under in vitro conditions. In other mammalian species, specific capacitation triggers have been described; unfortunately, none of these is able to induce full capacitation in stallion spermatozoa. Nevertheless, knowledge of capacitation pathways and their molecular triggers might improve our understanding of capacitation-related events observed in stallion sperm. When sperm cells are exposed to appropriate capacitation triggers, several molecular and biochemical changes should be induced in the sperm plasma membrane and cytoplasm. At the level of the sperm plasma membrane, (1) an increase in membrane fluidity, (2) cholesterol depletion and (3) lipid raft aggregation should occur consecutively; the cytoplasmic changes consist of protein tyrosine phosphorylation and elevated pH, cAMP and Ca2+ concentrations. These capacitation-related events enable the switch from progressive to hyperactivated motility of the sperm cells, and the induction of the acrosome reaction. These final capacitation triggers are indispensable for sperm cells to migrate through the viscous oviductal environment, penetrate the cumulus cells and zona pellucida and, finally, fuse with the oolemma. This review will focus on molecular aspects of sperm capacitation and known triggers in various mammalian species. Similarities and differences with the horse will be highlighted to improve our understanding of equine sperm capacitation/fertilizing events.

scholarly journals cDNA cloning reveals the molecular structure of a sperm surface protein, PH-20, involved in sperm-egg adhesion and the wide distribution of its gene among mammals.

1990 ◽  
Vol 111 (6) ◽  
pp. 2939-2949 ◽  
Author(s):  
W F Lathrop ◽  
E P Carmichael ◽  
D G Myles ◽  
P Primakoff
Keyword(s):  
Plasma Membrane ◽  
Guinea Pig ◽  
Zona Pellucida ◽  
Genomic Dna ◽  
Surface Protein ◽  
Wide Distribution ◽  
Cdna Clones ◽  
Southern Blots ◽  
Binding Domains ◽  
Acrosomal Membrane

Sperm binding to the egg zona pellucida in mammals is a cell-cell adhesion process that is generally species specific. The guinea pig sperm protein PH-20 has a required function in sperm adhesion to the zona pellucida of guinea pig eggs. PH-20 is located on both the sperm plasma membrane and acrosomal membrane. We report here the isolation and sequence of a full-length cDNA for PH-20 (available from EMBL/GenBank/DDBJ under accession number X56332). The derived amino acid sequence shows a mature protein of 468 amino acids containing six N-linked glycosylation sites and twelve cysteines, eight of which are tightly clustered near the COOH terminus. The sequence indicates PH-20 is a novel protein with no relationship to the mouse sperm adhesion protein galactosyl transferase and no significant homology with other known proteins. The two PH-20 populations, plasma membrane and acrosomal membrane, could arise because one form of PH-20 is encoded and differentially targeted at different spermatogenic stages. Alternatively, two different forms of PH-20 could be encoded. Our evidence thus far reveals only one sequence coding for PH-20: Southern blots of guinea pig genomic DNA indicated there is a single PH-20 gene, Northern blots showed a single size PH-20 message (approximately 2.2 kb), and no sequence variants were found among the sequenced cDNA clones. Cross-species Southern blots reveal the presence of a homologue of the PH-20 gene in mouse, rat, hamster, rabbit, bovine, monkey, and human genomic DNA, showing the PH-20 gene is conserved among mammals. Since genes for zona glycoproteins are also conserved among mammals, the general features of sperm and zona proteins involved in mammalian sperm-egg adhesion may have been evolutionarily maintained. Species specificity may result from limited changes in these molecules, either in their binding domains or in other regions that affect the ability of the binding domains to interact.

scholarly journals Establishment of the mammalian membrane block to polyspermy: evidence for calcium-dependent and -independent regulation

Reproduction ◽  
2007 ◽  
Vol 133 (2) ◽  
pp. 383-393 ◽  
Author(s):  
Allison J Gardner ◽  
Carmen J Williams ◽  
Janice P Evans
Keyword(s):  
Plasma Membrane ◽  
Zona Pellucida ◽  
Egg Activation ◽  
Membrane Function ◽  
Independent Event ◽  
Calcium Dependent ◽  
Normal Sperm ◽  
Egg Membrane ◽  
Mammalian Eggs ◽  
Mouse Eggs

One crucial result of egg activation is the establishment of blocks on the zona pellucida and the egg plasma membrane to prevent fertilization by additional sperm. The mechanism(s) by which a mammalian egg regulates the establishment of the membrane block to polyspermy is largely unknown. Since Ca2+ signaling regulates several egg activation events, this study investigates how sperm-induced Ca2+ transients affect the membrane block to polyspermy, building on our previous work (Biology of Reproduction 67:1342). We demonstrate that mouse eggs that experience only one sperm-induced Ca2+ transient establish a membrane block that is less effective, than in eggs that experience normal sperm-induced Ca2+ transients but that is more effective than in eggs with completely suppressed [Ca2+]cyt increases. Sperm-induced increases in [Ca2+]cyt regulate the timing of membrane block establishment, as this block is established more slowly in eggs that experience one or no sperm-induced Ca2+ transients. Finally, our studies produce the intriguing discovery that there is also a Ca2+-independent event that is associated with fertilization in the pathway leading to membrane block establishment. Taken together, these data indicate that Ca2+ plays a role in facilitating membrane block establishment by regulating the timing with which this change in egg membrane function occurs, and also that the membrane block differs from other post-fertilization egg activation responses as Ca2+ is not the only stimulus. The membrane block to polyspermy in mammalian eggs is likely to be the culmination of multiple post-fertilization events that together modify the egg membrane’s receptivity to sperm.

scholarly journals Role of actin cytoskeleton in mammalian sperm capacitation and the acrosome reaction

Reproduction ◽  
2005 ◽  
Vol 129 (3) ◽  
pp. 263-268 ◽  
Author(s):  
Haim Breitbart ◽  
Gili Cohen ◽  
Sara Rubinstein
Keyword(s):  
Plasma Membrane ◽  
Zona Pellucida ◽  
Acrosome Reaction ◽  
Reproductive Tract ◽  
Female Reproductive Tract ◽  
Mammalian Sperm ◽  
Acrosomal Membrane ◽  
Close Proximity ◽  
Sperm Plasma Membrane ◽  
To Come

In order to fertilize, the mammalian spermatozoa should reside in the female reproductive tract for several hours, during which they undergo a series of biochemical modifications collectively called capacitation. Only capacitated sperm can undergo the acrosome reaction after binding to the egg zona pellucida, a process which enables sperm to penetrate into the egg and fertilize it. Polymerization of globular (G)-actin to filamentous (F)-actin occurs during capacitation, depending on protein kinase A activation, protein tyrosine phosphorylation, and phospholipase D activation. F-actin formation is important for the translocation of phospholipase C from the cytosol to the sperm plasma membrane during capacitation. Prior to the occurrence of the acrosome reaction, the F-actin should undergo depolymerization, a necessary process which enables the outer acrosomal membrane and the overlying plasma membrane to come into close proximity and fuse. The binding of the capacitated sperm to the zona pellucida induces a fast increase in sperm intracellular calcium, activation of actin severing proteins which break down the actin fibers, and allows the acrosome reaction to take place.

Distribution of VLA-5 and VLA-4 fibronectin receptors in human monocytes demonstrated by immunofluorescence, immunocytochemistry, and autoradiography

1993 ◽  
Vol 51 ◽  
pp. 306-307
Author(s):  
Robert Williams ◽  
Che-Hung Lee ◽  
Sara E. Quella ◽  
David M. Harlan ◽  
Yuan-Hsu Kang
Keyword(s):  
Present Report ◽  
Matrix Proteins ◽  
Extracellular Matrices ◽  
Human Monocytes ◽  
Integrin Receptors ◽  
Morphological Evaluation ◽  
The Matrix ◽  
Specific Receptors ◽  
Cytokine Stimulation ◽  
Monocyte Adherence

Monocyte adherence to endothelial or extracellular matrices plays an important role in triggering monocyte activation in extravascular sites of infection, chronic inflammatory disorders, and tissue damage. Migration of monocytes in the tissues involves the response to a chemoattractant and movement by a series of attachments and detachments to the extracellular matrices which are regulated by expression and distribution of specific receptors for the matrix proteins such as fibronectin (FN). The VSAs (very late antigens or beta integrins), a subfamily of the transmembrane heterodimeric integrin receptors, have been thought to play a major role in monocyte adherence to the extracellular matrices and cells. In this subfamily, VLA-5 and VLA-4 are believed to be the most essential integrins mediating monocyte adherence to FN. In the present report, we have established and compared different procedures for morphological evaluation of the expression and distribution of the FN receptors on human monocytes in order to investigate their response to endotoxin or cytokine stimulation.

scholarly journals Pig zona pellucida 2 (pZP2) protein does not participate in zona pellucida formation in transgenic mice

Reproduction ◽  
2006 ◽  
Vol 132 (3) ◽  
pp. 455-464 ◽  
Author(s):  
Akiko Hasegawa ◽  
Nozomi Kanazawa ◽  
Hideaki Sawai ◽  
Shinji Komori ◽  
Koji Koyama
Keyword(s):  
Extracellular Matrix ◽  
Transgenic Mice ◽  
Molecular Species ◽  
Zona Pellucida ◽  
Transgenic Protein ◽  
Specific Manner ◽  
Species Specific ◽  
Deficient Mice

The zona pellucida, an extracellular matrix surrounding mammalian oocytes, is composed of three or four glycoproteins. It is well known that the zona pellucida plays several critical roles during fertilization, but there is little knowledge about its formation. The purpose of this study is to examine whether a pig zona pellucida glycoprotein 2 (pZP2) would assemble with mouse zona pellucida. A transgene construct was prepared by placing a minigene encoding pZP2 downstream from the promoter of mouse ZP2. The result showed that the transgenic protein was synthesized in growing oocytes but not incorporated into the zona pellucida. Furthermore, the pZP2 transgene did not rescue the phenotype in ZP2-knockout zona-deficient mice. These results indicate that pZP2 does not participate in mouse zona pellucida formation and the zona pellucida is constituted from its component proteins in a molecular species-specific manner between mice and pigs.

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