scholarly journals Phospholipase Cδ4 is required for Ca2+ mobilization essential for acrosome reaction in sperm

2003 ◽  
Vol 161 (1) ◽  
pp. 79-88 ◽  
Author(s):  
Kiyoko Fukami ◽  
Manabu Yoshida ◽  
Takafumi Inoue ◽  
Manabu Kurokawa ◽  
Rafael A. Fissore ◽  
...  
Keyword(s):  
Zona Pellucida ◽  
Calcium Imaging ◽  
Acrosome Reaction ◽  
Sperm Head ◽  
Wild Type ◽  
Imaging Studies ◽  
Mouse Sperm ◽  
Mammalian Fertilization ◽  
Important Enzyme

Zona pellucida (ZP)–induced acrosome reaction in sperm is a required step for mammalian fertilization. However, the precise mechanism of the acrosome reaction remains unclear. We previously reported that PLCδ4 is involved in the ZP-induced acrosome reaction in mouse sperm. Here we have monitored Ca2+ responses in single sperm, and we report that the [Ca2+]i increase in response to ZP, which is essential for driving the acrosome reaction in vivo, is absent in PLCδ4−/− sperm. Progesterone, another physiological inducer of the acrosome reaction, failed to induce sustained [Ca2+]i increases in PLCδ4−/− sperm, and consequently the acrosome reaction was partially inhibited. In addition, we observed oscillatory [Ca2+]i increases in wild-type sperm in response to these acrosome inducers. Calcium imaging studies revealed that the [Ca2+]i increases induced by exposure to ZP and progesterone started at different sites within the sperm head, indicating that these agonists induce the acrosome reaction via different Ca2+ mechanisms. Furthermore, store-operated channel (SOC) activity was severely impaired in PLCδ4−/− sperm. These results indicate that PLCδ4 is an important enzyme for intracellular [Ca2+]i mobilization in the ZP-induced acrosome reaction and for sustained [Ca2+]i increases through SOC induced by ZP and progesterone in sperm.

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 Mouse gamete interactions during fertilization in vitro. Chlortetracycline as a fluorescent probe for the mouse sperm acrosome reaction.

1979 ◽  
Vol 83 (3) ◽  
pp. 544-555 ◽  
Author(s):  
P M Saling ◽  
B T Storey
Keyword(s):  
Plasma Membrane ◽  
Fluorescent Probe ◽  
Zona Pellucida ◽  
Acrosome Reaction ◽  
Sperm Head ◽  
Fertilization In Vitro ◽  
Ionophore A23187 ◽  
Mouse Sperm ◽  
Sperm Acrosome

We have developed an assay for detecting the acrosome reaction in mouse sperm using chlortetracycline (CTC) as a fluorescent probe. Sperm known to be intact with nonreacted acrosomes show CTC fluorescence in the presence of Ca2+ over the anterior portion of the sperm head on the plasma membrane covering the acrosome. Sperm which have undergone the acrosome reaction do not show fluorescence on the sperm head. Mouse sperm bind to zonae pellucidae of cumulus-free eggs in vitro in a Ca2+-dependent reaction; these sperm are intact by the CTC assay. Intact sperm bind to mechanically isolated zonae under the same conditions: the egg is apparently unnecessary for this inital reaction. Sperm suspensions, in which greater than 50% of the motile population had completed the acrosome reaction, were prepared by incubation in hyperosmolal medium followed by treatment with the divalent cation ionophore, A23187. Cumulus-free eggs challenged with such sperm suspensions preferentially bind intact sperm; acrosome-reacted sperm do not bind. We conclude that the plasma membrane of the mouse sperm is responsible for recognition of the egg's zona pellucida and that the obligatory sequence of reactions leading to fusion of mouse gametes is binding of the intact sperm to the zona pellucida, followed by the acrosome reaction at the zona surface, followed in turn by sperm penetration of the zona.

scholarly journals The sperm protein SPACA4 is required for efficient fertilization in mice

2021 ◽  
Author(s):  
Sarah Herberg ◽  
Yoshitaka Fujihara ◽  
Andreas Blaha ◽  
Karin Panser ◽  
Kiyonari Kobayashi ◽  
...  
Keyword(s):  
Zona Pellucida ◽  
Knockout Mice ◽  
Mammalian Species ◽  
Wild Type ◽  
Sperm Protein ◽  
Mammalian Sperm ◽  
Fundamental Process ◽  
Mammalian Fertilization

Fertilization is the fundamental process that initiates the development of a new individual in all sexually reproducing species. Despite its importance, our understanding of the molecular players that govern mammalian sperm-egg interaction is incomplete, partly because many of the essential factors found in non-mammalian species do not have obvious mammalian homologs. We have recently identified the Ly6/uPAR protein Bouncer as a new, essential fertilization factor in zebrafish (Herberg et al., 2018). Here, we show that Bouncer's homolog in mammals, SPACA4, is also required for efficient fertilization in mice. In contrast to fish, where Bouncer is expressed specifically in the egg, SPACA4 is expressed exclusively in the testis. Male knockout mice are severely sub-fertile, and sperm lacking SPACA4 fail to fertilize wild-type eggs in vitro. Interestingly, removal of the zona pellucida rescues the fertilization defect of Spaca4-deficient sperm in vitro, indicating that SPACA4 is not required for the interaction of sperm and the oolemma but rather of sperm and zona pellucida. Our work identifies SPACA4 as an important sperm protein necessary for zona pellucida penetration during mammalian fertilization.

scholarly journals A single domain of the ZP2 zona pellucida protein mediates gamete recognition in mice and humans

2014 ◽  
Vol 205 (6) ◽  
pp. 801-809 ◽  
Author(s):  
Matteo A. Avella ◽  
Boris Baibakov ◽  
Jurrien Dean
Keyword(s):  
Transgenic Mice ◽  
Zona Pellucida ◽  
Human Sperm ◽  
Recombinant Peptide ◽  
Genetic Ablation ◽  
Sperm Binding ◽  
Gamete Recognition ◽  
Mammalian Fertilization ◽  
Human And Mouse

The extracellular zona pellucida surrounds ovulated eggs and mediates gamete recognition that is essential for mammalian fertilization. Zonae matrices contain three (mouse) or four (human) glycoproteins (ZP1–4), but which protein binds sperm remains controversial. A defining characteristic of an essential zona ligand is sterility after genetic ablation. We have established transgenic mice expressing human ZP4 that form zonae pellucidae in the absence of mouse or human ZP2. Neither mouse nor human sperm bound to these ovulated eggs, and these female mice were sterile after in vivo insemination or natural mating. The same phenotype was observed with truncated ZP2 that lacks a restricted domain within ZP251–149. Chimeric human/mouse ZP2 isoforms expressed in transgenic mice and recombinant peptide bead assays confirmed that this region accounts for the taxon specificity observed in human–mouse gamete recognition. These observations in transgenic mice document that the ZP251–149 sperm-binding domain is necessary for human and mouse gamete recognition and penetration through the zona pellucida.

scholarly journals Epididymal Maturation and the Acrosome Reaction in Mouse Sperm: Response to Zona Pellucida Develops Coincident with Modification of M42 Antigen1

1988 ◽  
Vol 38 (1) ◽  
pp. 221-233 ◽  
Author(s):  
Katherine A. Lakoski ◽  
Christopher P. Carron ◽  
Christine L. Cabot ◽  
Patricia M. Saling
Keyword(s):  
Zona Pellucida ◽  
Acrosome Reaction ◽  
Mouse Sperm ◽  
Epididymal Maturation

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.

Effect of Epidermal Growth Factor on Mouse Sperm Acrosome Reaction Induced by Zona Pellucida

1994 ◽  
Vol 31 (2-3) ◽  
pp. 116-122 ◽  
Author(s):  
Satoru Furuya ◽  
Yoshihiro Endo ◽  
Mikiko Oba ◽  
Yukari Matsui ◽  
Shuetu Suzuki ◽  
...  
Keyword(s):  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Zona Pellucida ◽  
Acrosome Reaction ◽  
Mouse Sperm ◽  
Epidermal Growth ◽  
Sperm Acrosome Reaction ◽  
Sperm Acrosome
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