Delineation of downstream signalling components during acrosome reaction mediated by heat solubilized human zona pellucida

Bhandari, Beena; Bansal, Pankaj; Talwar, Pankaj; Gupta, Seema

doi:10.1186/1477-7827-8-7

Cited by 17 publications

(20 citation statements)

References 45 publications

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“…The acrosome reaction mediated by human SIZP involves activation of both a G i protein-coupled receptor (Lee et al 1992;Franken et al 1996;Bastiaan et al 1999;Bhandari et al 2010) as well as TK (Bhandari et al 2010). Among the downstream effectors, human SIZP also involves activation of protein kinase A (PKA), protein kinase C (PKC) and phosphoinositide 3 (PI3)-kinase (Fisher et al 1998;Liguori et al 2005;Bhandari et al 2010) to mediate the acrosome reaction.…”

Section: Signaling Events Associated With the Acrosome-reaction Mediamentioning

confidence: 97%

“…However, recent observations suggest that L-type VOCCs also play an important role in the human ZP3-mediated increase in [Ca 2+ ] i and thereby acrosome reaction (Jose et al 2010), whereas ZP1 and ZP4 employ both L-type and T-type VOCCs for [Ca 2+ ] i influx and acrosome reaction (Chiu et al 2008a;Ganguly et al 2010b). Extracellular Ca 2+ is critical for induction of the acrosome reaction by any of the above agonists, as pre-incubation with the calcium chelator, EGTA, decreases the acrosome reaction to the levels of a spontaneous acrosome reaction (Schuffner et al 2002;Chiu et al 2008a;Bhandari et al 2010;Ganguly et al 2010b). IP 3 may further facilitate an increase in [Ca 2+ ] i release through modulation of IP 3 -sensitive intracellular calcium stores (Walensky and Snyder, 1995).…”

Section: Signaling Events Associated With the Acrosome-reaction Mediamentioning

confidence: 98%

“…Incubation of capacitated human sperm with intact human zonae pellucidae, or SIZP obtained either by acid disaggregation or heat, led to a significant increase in the acrosome reaction (Cross et al 1988;Bhandari et al 2010).…”

Section: Structural and Functional Aspects Of Zp Glycoproteins Duringmentioning

confidence: 98%

“…Among the downstream effectors, human SIZP also involves activation of protein kinase A (PKA), protein kinase C (PKC) and phosphoinositide 3 (PI3)-kinase (Fisher et al 1998;Liguori et al 2005;Bhandari et al 2010) to mediate the acrosome reaction. A SIZP-mediated acrosome reaction involves predominantly T-type VOCCs leading to an increase in the [Ca 2+ ] i -a hallmark for induction of the acrosome reaction-and also uses GABA-A receptorassociated chloride (Cl − ) channels (Breitbart 2002;Patrat et al 2006;Bhandari et al 2010).…”

Section: Signaling Events Associated With the Acrosome-reaction Mediamentioning

confidence: 99%

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Mammalian zona pellucida glycoproteins: structure and function during fertilization

et al. 2012

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Zona pellucida (ZP) is a glycoproteinaceous translucent matrix that surrounds the mammalian oocyte and plays a critical role in the accomplishment of fertilization. In humans, it is composed of 4 glycoproteins designated as ZP1, ZP2, ZP3 and ZP4, whereas mouse ZP is composed of ZP1, ZP2 and ZP3 (Zp4 being a pseudogene). In addition to a variable sequence identity of a given zona protein among various species, human ZP1 and ZP4 are paralogs and mature polypeptide chains share an identity of 47%. Employing either affinity purified native or recombinant human zona proteins, it has been demonstrated that ZP1, ZP3 and ZP4 bind to the capacitated human spermatozoa and induce an acrosome reaction, whereas in mice, ZP3 acts as the putative primary sperm receptor. Human ZP2 only binds to acrosome-reacted spermatozoa and thus may be acting as a secondary sperm receptor. In contrast to O-linked glycans of ZP3 in mice, N-linked glycans of human ZP3 and ZP4 are more relevant for induction of the acrosome reaction. Recent studies suggest that Sialyl-Lewis(x) sequence present on both N- and O-glycans of human ZP play an important role in human sperm-egg binding. There are subtle differences in the downstream signaling events associated with ZP3 versus ZP1/ZP4-mediated induction of the acrosome reaction. For example, ZP3 but not ZP1/ZP4-mediated induction of the acrosome reaction is dependent on the activation of the Gi protein-coupled receptor. Thus, various studies suggest that, in contrast to mice, in humans more than one zona protein binds to spermatozoa and induces an acrosome reaction.

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Section: Signaling Events Associated With the Acrosome-reaction Mediamentioning

confidence: 97%

Section: Signaling Events Associated With the Acrosome-reaction Mediamentioning

confidence: 98%

Section: Structural and Functional Aspects Of Zp Glycoproteins Duringmentioning

confidence: 98%

Section: Signaling Events Associated With the Acrosome-reaction Mediamentioning

confidence: 99%

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Mammalian zona pellucida glycoproteins: structure and function during fertilization

et al. 2012

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“…39,40 The high intracellular free calcium concentration together with DAG leads to membrane fusion and finally acrosomal exocytosis. 2,41 Induction of the AR by the solubilized human ZP depends on extracellular Ca 2120, 42 and involves activation of G i protein-coupled receptor pathway signalling, 20,[42][43][44][45] tyrosine kinases, 42 PKA, PKC, phosphoinositide-3 kinase, 42,46 T-type VOCCs and gamma aminobutyric acid-A receptor-associated chloride channels. 42 …”

Section: Signalling Events During Zp-mediated Acrosomal Exocytosismentioning

confidence: 99%

Acrosome reaction: relevance of zona pellucida glycoproteins

Gupta

Bhandari²

2010

Asian J Androl

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During mammalian fertilisation, the zona pellucida (ZP) matrix surrounding the oocyte is responsible for the binding of the spermatozoa to the oocyte and induction of the acrosome reaction (AR) in the ZP-bound spermatozoon. The AR is crucial for the penetration of the ZP matrix by spermatozoa. The ZP matrix in mice is composed of three glycoproteins designated ZP1, ZP2 and ZP3, whereas in humans, it is composed of four (ZP1, ZP2, ZP3 and ZP4). ZP3 acts as the putative primary sperm receptor and is responsible for AR induction in mice, whereas in humans (in addition to ZP3), ZP1 and ZP4 also induce the AR. The ability of ZP3 to induce the AR resides in its C-terminal fragment. O-linked glycans are critical for the murine ZP3-mediated AR. However, N-linked glycans of human ZP1, ZP3 and ZP4 have important roles in the induction of the AR. Studies with pharmacological inhibitors showed that the ZP3-induced AR involves the activation of the G i -coupled receptor pathway, whereas ZP1-and ZP4-mediated ARs are independent of this pathway. The ZP3-induced AR involves the activation of T-type voltage-operated calcium channels (VOCCs), whereas ZP1-and ZP4-induced ARs involve both T-and L-type VOCCs. To conclude, in mice, ZP3 is primarily responsible for the binding of capacitated spermatozoa to the ZP matrix and induction of the AR, whereas in humans (in addition to ZP3), ZP1 and ZP4 also participate in these stages of fertilisation. Keywords: acrosome reaction; fertilisation; oocyte; signalling pathways; spermatozoa; zona pellucida glycoproteins INTRODUCTION Mammalian fertilisation is a highly synchronized process that involves a complex series of interactions between the spermatozoon and the egg, culminating in their unison. The initial steps in fertilisation involves the binding of the spermatozoon to the zona pellucida (ZP) matrix surrounding the egg, followed by induction of the acrosome reaction (AR) in the zona-bound spermatozoon, a pre-requisite for penetration of the ZP matrix by the spermatozoon. The spermatozoon acrosome is a Golgi-derived organelle that forms a cap over the anterior two-thirds of its nucleus. The AR involves the fusion of the sperm membrane with the outer acrosomal membrane, resulting in release of the acrosomal contents and exposure of the inner acrosomal membrane on the anterior head of the spermatozoon. Various physiological agents, such as progesterone, serum albumin, follicular fluid, hormones (including biogenic amines), hydrolytic enzymes (particularly proteases), hyaluronic acid and ZP glycoproteins, have been implicated in the induction of the AR. [1][2][3] In the present paper, we review the role of the ZP matrix and its constituents in AR induction. Various downstream signalling pathways involved in the ZP glycoprotein-induced AR will also be discussed. On the basis of the current literature and studies from our group, the salient differences in the ZP glycoprotein-mediated induction of the AR in mouse versus human will be highlighted.

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Sperm‐Binding Assay Using an In Vitro 3D Model of the Mammalian Cumulus‐Oocyte Complex

2020

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We have recently described a new model to study gamete interaction in mammalian species. The model recreates the spherical surface of the oocyte by using magnetic Sepharose beads coated with a layer of a recombinant protein involved in gamete interaction (such as ZP2, or the IZUMO1 receptor JUNO) and an external layer of cumulus oophorus cells, thus mimicking, to some extent, a native cumulus‐oocyte complex. Once generated, this 3D model can be used in a sperm‐binding assay to obtain valuable information about the molecular basis of gamete interaction, since different recombinant proteins can be used to coat the bead surface, thus generating a variety of models to be used for several species. Furthermore, thanks to the ability of the model to decoy sperm, the physiological status of the bound sperm can be studied, making this a powerful tool to select sperm with high fertilizing capacity, to unmask subfertile animals in livestock breeding centers, or for toxicological studies. Here, we describe how to generate and use this model for sperm‐binding assays, using porcine sperm as an example, and ZP2, a protein from zona pellucida, as the recombinant protein of interest. © 2020 Wiley Periodicals LLC. Basic Protocol 1: Generation of the in vitro 3D model Alternate Protocol 1: Binding cumulus oophorus cells to the model Basic Protocol 2: Quality control of the model by SDS‐PAGE electrophoresis and western blot Support Protocol 1: Immunochemistry to confirm proper protein distribution on surface of beads Support Protocol 2: Elution of recombinant conjugated proteins Basic Protocol 3: Sperm‐binding assay Alternate Protocol 2: Sperm preparation by the swim‐up method

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Delineation of downstream signalling components during acrosome reaction mediated by heat solubilized human zona pellucida

Cited by 17 publications

References 45 publications

Mammalian zona pellucida glycoproteins: structure and function during fertilization

Mammalian zona pellucida glycoproteins: structure and function during fertilization

Acrosome reaction: relevance of zona pellucida glycoproteins

Sperm‐Binding Assay Using an In Vitro 3D Model of the Mammalian Cumulus‐Oocyte Complex

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