Capacitation Induces Tyrosine Phosphorylation of Proteins in the Boar Sperm Plasma Membrane

Flesch, Frits M.; Colenbrander, B.; Golde, L.M.G. Van; Gadella, B.M.

doi:10.1006/bbrc.1999.1300

Cited by 145 publications

(89 citation statements)

References 25 publications

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“…2C). The possibility of a relationship between plasma membrane phosphoprotein expression and zona binding has been alluded to in other mammalian species (Flesch et al, 1999;Flesch et al, 2001;Tardif et al, 2001). However, this is the first report of the appearance of tyrosinephosphorylated proteins on the surface of live mouse spermatozoa during capacitation.…”

Section: Discussionmentioning

confidence: 94%

Tyrosine phosphorylation activates surface chaperones facilitating sperm-zona recognition

Asquith

Baleato

McLaughlin

et al. 2004

Journal of Cell Science

182

183

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Mammalian spermatozoa undergo a series of molecular and biochemical changes collectively termed capacitation prior to acquiring the ability to fertilise the oocyte. Although phosphorylation of sperm proteins on tyrosine residues has been recognised as an important component of this process, the precise relationship between the phosphorylation status of mammalian spermatozoa and their capacity for fertilisation has remained unclear. In this study we demonstrate a causal relationship between tyrosine phosphorylation in spermatozoa and sperm-zona interaction. The phosphotyrosine expression associated with sperm capacitation localised to internal flagellar structures in permeabilised cells but could also be detected on the exterior surface of the sperm head in live cells. Importantly, almost all spermatozoa bound to the zona pellucida demonstrated this pattern of phosphoprotein localisation, compared to fewer than 15% of the free-swimming population. These data suggest that tyrosine phosphorylation plays a significant role in remodelling the sperm surface, so that these cells are able to recognise the zona pellucida. Phosphoproteome analysis yielded the first evidence of molecular chaperones, endoplasmin (erp99) and heat shock protein 60 (hsp60), as targets for phosphorylation on the surface of mouse spermatozoa, whereas immunofluorescence localised these proteins to the precise region of the sperm head that participates in zona recognition. Based on these results, we propose a novel mechanism for mammalian gamete interaction whereby the activation of sperm-surface chaperones by tyrosine phosphorylation during capacitation may trigger conformational changes facilitating the formation of a functional zona pellucida receptor complex on the surface of mammalian spermatozoa.

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Section: Discussionmentioning

confidence: 94%

Tyrosine phosphorylation activates surface chaperones facilitating sperm-zona recognition

Asquith

Baleato

McLaughlin

et al. 2004

Journal of Cell Science

182

183

View full text Add to dashboard Cite

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“…Both samples were randomly divided into three groups (n ϭ 3) for experimental replication. To rule out individual variation, each group was pooled, and pooled samples were washed at 500 ϫ g for 20 min with a discontinuous (70% [v/v] and 35% [v/v]) Percoll gradient (Sigma, St Louis, MO) to remove seminal plasma and dead spermatozoa (21).…”

Section: Methodsmentioning

confidence: 99%

Discovery of Predictive Biomarkers for Litter Size in Boar Spermatozoa*

Kwon

Rahman

Lee

et al. 2015

Molecular & Cellular Proteomics

124

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Conventional semen analysis has been used for prognosis and diagnosis of male fertility. Although this tool is essential for providing initial quantitative information about semen, it remains a subject of debate. Therefore, development of new methods for the prognosis and diagnosis of male fertility should be seriously considered for animal species of economic importance as well as for humans. In the present study, we applied a comprehensive proteomic approach to identify global protein biomarkers in boar spermatozoa in order to increase the precision of male fertility prognoses and diagnoses. We determined that L-amino acid oxidase, mitochondrial malate dehydrogenase 2, NAD (MDH2), cytosolic 5-nucleotidase 1B, lysozyme-like protein 4, and calmodulin (CALM) were significantly and abundantly expressed in high-litter size spermatozoa. We also found that equatorin, spermadhesin AWN, triosephosphate isomerase (TPI), Ras-related protein Rab-2A (RAB2A), spermadhesin AQN-3, and NADH dehydrogenase [ubiquinone] iron-sulfur protein 2 (NDUFS2) were significantly and abundantly expressed in low-litter size spermatozoa (>3-fold). Moreover, RAB2A, TPI, and NDUFS2 were negatively correlated with litter size, whereas CALM and MDH2 were positively correlated. This study provides novel biomarkers for the prediction of male fertility. To the best of our knowledge, this is the first work that shows significantly increased litter size using male fertility biomarkers in a field trial. Moreover, these protein markers may provide new developmental tools for the selection of superior sires as well as for the prognosis and diagnosis of male fertility.

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“…However, a cAMP-dependent increase in 32-kDa tyrosine-phosphorylated protein (called TyrP32 or p32) in boar spermatozoa is exceptionally suppressed by inhibition of tyrosine phosphatases and barely affected by inhibition of protein tyrosine kinases [6]. This tyrosine-phosphorylated protein is primarily found as a capacitation-related protein [7][8][9] and has recently been identified as a (pro)acrosin-binding protein [10,11]. We have previously shown that the cAMP-dependent increase in TyrP32 is greater in samples containing many spermatozoa that undergo calcium-dependent changes in acrosomal morphology (acrosome reaction) [6].…”

mentioning

confidence: 99%

A 32-kDa Tyrosine-phosphorylated Protein Shows a Protease-dependent Increase in Dead Boar Spermatozoa

Tabuchi

Shidara

Harayama

2008

Journal of Reproduction and Development

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Abstract. Boar sperm TyrP32 is a 32-kDa tyrosine-phosphorylated protein that increases during the capacitation and acrosome reaction and during cryocapacitation. However, it is still unclear whether the increase in TyrP32 is an event that is limited to the process of sperm fertilization, including cryocapacitation. The aims of the present study were to demonstrate that TyrP32 is increased in dead spermatozoa after freeze-thawing without a cryoprotectant and to find the causal factors for this increase. Washed spermatozoa were resuspended in a salt solution and then frozen. The frozen samples were rapidly thawed in a warm water bath and then used for sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE)/Western blotting to detect TyrP32, SDS-PAGE/silver staining of sperm proteins and staining of acrosomal contents with fluorescein isothiocyanate (FITC)-conjugated peanut agglutinin (PNA). In the samples before freezing, TyrP32 was barely detectable, and the distribution of the acrosomal contents was normal in most spermatozoa. One cycle of freeze-thawing induced an increase in TyrP32, a decrease in major sperm proteins and disorder in the acrosomal contents. However, the addition of a protease inhibitor (APMSF, 1 mM) suppressed the increase in TyrP32 and the decrease in the major sperm proteins, although it did not have any influence on the disorder in the acrosomal contents. Additionally, the spermatozoa did not exhibit any flagellar movement after freeze-thawing, which showed that almost all of them were dead. These results indicate that TyrP32 can show a protease-dependent increase in dead spermatozoa after freeze-thawing without a cryoprotectant even though the dead spermatozoa do not undergo cryocapacitation. Key words: Acrosome, Boar, Peanut agglutinin (PNA), Protease, Sperm, Tyrosine (J. Reprod. Dev. 54: [502][503][504][505][506][507] 2008) n mammalian spermatozoa, a variety of tyrosine-phosphorylated proteins increase during incubation under capacitationsupporting conditions. These proteins have been considered to be involved in regulation of fertilization-related events [1,2]. It is generally believed that the increase in tyrosine-phosphorylated proteins results from activation of protein tyrosine kinases and inactivation of protein tyrosine phosphatases that are controlled by cyclic adenosine 3',5'-monophosphate (cAMP)/protein kinase A (PKA) signaling [1,[3][4][5]. However, a cAMP-dependent increase in 32-kDa tyrosine-phosphorylated protein (called TyrP32 or p32) in boar spermatozoa is exceptionally suppressed by inhibition of tyrosine phosphatases and barely affected by inhibition of protein tyrosine kinases [6]. This tyrosine-phosphorylated protein is primarily found as a capacitation-related protein [7][8][9] and has recently been identified as a (pro)acrosin-binding protein [10,11]. We have previously shown that the cAMP-dependent increase in TyrP32 is greater in samples containing many spermatozoa that undergo calcium-dependent changes in acrosomal morphology (acrosome reaction) ...

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Capacitation Induces Tyrosine Phosphorylation of Proteins in the Boar Sperm Plasma Membrane

Cited by 145 publications

References 25 publications

Tyrosine phosphorylation activates surface chaperones facilitating sperm-zona recognition

Tyrosine phosphorylation activates surface chaperones facilitating sperm-zona recognition

Discovery of Predictive Biomarkers for Litter Size in Boar Spermatozoa*

A 32-kDa Tyrosine-phosphorylated Protein Shows a Protease-dependent Increase in Dead Boar Spermatozoa

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