Effect of sex sorting on stallion spermatozoa: Heterologous oocyte binding, tyrosine phosphorylation and acrosome reaction assay

Silva, C Balao da; Spinaci, Marcella; Bucci, Diego; Giaretta, Elisa; Peña, Fernando J.; Mari, Gaetano; Galeati, Giovanna

doi:10.1016/j.anireprosci.2013.07.008

Cited by 15 publications

(21 citation statements)

References 44 publications

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“…However, equine sperm-bovine ZP binding can be functional, as acrosome reaction and penetration through bovine ZP have been shown to take place (Sinowatz et al 2003). In our experiments, we observed an individual variability among stallions in the number of sperm attached to bovine ZP, as described previously (Balao da Silva et al 2013). Additionally, incubation with MbCD did not affect sperm binding in two stallions, but in one stallion (IP), incubation with 1 mM MbCD in MWCCC increased sperm binding over MWCCC alone.…”

Section: Discussionsupporting

confidence: 84%

Effect of calcium, bicarbonate, and albumin on capacitation-related events in equine sperm

Macías‐García¹,

González‐Fernández²,

Loux³

et al. 2015

Reproduction

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Repeatable methods for IVF have not been established in the horse, reflecting the failure of standard capacitating media to induce changes required for fertilization capacity in equine sperm. One important step in capacitation is membrane cholesterol efflux, which in other species is triggered by cholesterol oxidation and is typically enhanced using albumin as a sterol acceptor. We incubated equine sperm in the presence of calcium, BSA, and bicarbonate, alone or in combination. Bicarbonate induced an increase in reactive oxygen species (ROS) that was abolished by the addition of calcium or BSA. Bicarbonate induced protein tyrosine phosphorylation (PY), even in the presence of calcium or BSA. Incubation at high pH enhanced PY but did not increase ROS production. Notably, no combination of these factors was associated with significant cholesterol efflux, as assessed by fluorescent quantitative cholesterol assay and confirmed by filipin staining. By contrast, sperm treated with methyl-b-cyclodextrin showed a significant reduction in cholesterol levels, but no significant increase in PY or ROS. Presence of BSA increased sperm binding to bovine zonae pellucidae in all three stallions. These results show that presence of serum albumin is not associated with a reduction in membrane cholesterol levels in equine sperm, highlighting the failure of equine sperm to exhibit core capacitation-related changes in a standard capacitating medium. These data indicate an atypical relationship among cholesterol efflux, ROS production, and PY in equine sperm. Our findings may help to elucidate factors affecting failure of equine IVF under standard conditions.

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Section: Discussionsupporting

confidence: 84%

Effect of calcium, bicarbonate, and albumin on capacitation-related events in equine sperm

Macías‐García¹,

González‐Fernández²,

Loux³

et al. 2015

Reproduction

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“…; Balao da Silva et al. ). As in the case of the homologous assay, the process of capacitation is needed for stallion spermatozoa to bind to heterologous oocytes (Clulow et al.…”

Section: Discussionmentioning

confidence: 98%

Epigallocatechin‐3‐Gallate (EGCG) Reduces Rotenone Effect on Stallion Sperm–Zona Pellucida Heterologous Binding

Dávila

Bucci

Galeati

et al. 2015

Reprod Domestic Animals

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Stallion spermatozoa are highly dependent on oxidative phosphorylation for ATP production to achieve normal sperm function and to fuel the motility. The aim of this study was to evaluate the response of equine sperm under capacitating conditions to the inhibition of mitochondrial complex I by rotenone and to test whether epigallocatechin-3-gallate (EGCG), a natural polyphenol component of green tea, could counteract this effect. After 2-h incubation of stallion spermatozoa in modified Tyrode's medium, rotenone (100 nm, 500 nm and 5 μm) and EGCG (10, 20 and 60 μm), alone or in combination, did not induce any significant difference on the percentage of viable cells, live sperm with active mitochondria and spermatozoa with intact acrosome. The inhibition of complex I of mitochondrial respiratory chain of stallion sperm with rotenone exerted a negative effect on heterologous ZP binding ability. EGCG at the concentrations of 10 and 20 μm (but not of 60 μm) induced a significant increase in the number of sperm bound to the ZP compared with that for control. Moreover, when stallion sperm were treated with rotenone 100 nm, the presence of EGCG at all the concentrations tested (10, 20 and 60 μm) significantly increased the number of sperm bound to the ZP up to control levels, suggesting that this green tea polyphenol is able to reduce the toxicity of rotenone.

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“…Sex-sorting induces capacitation-like changes to sperm [10,38], with the removal of many beneficial seminal plasma components resulting in membrane destabilisation and greater acrosome reactivity [39]. Optimal insemination timing becomes increasingly important when sperm longevity has been compromised by exposure to stressors such as liquid storage, cryopreservation and sex-sorting.…”

Section: Discussionmentioning

confidence: 99%

Field fertility of liquid stored and cryopreserved flow cytometrically sex‐sorted stallion sperm

Gibb

Grupen

Maxwell

et al. 2016

Equine Veterinary Journal

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Effect of sex sorting on stallion spermatozoa: Heterologous oocyte binding, tyrosine phosphorylation and acrosome reaction assay

Cited by 15 publications

References 44 publications

Effect of calcium, bicarbonate, and albumin on capacitation-related events in equine sperm

Effect of calcium, bicarbonate, and albumin on capacitation-related events in equine sperm

Epigallocatechin‐3‐Gallate (EGCG) Reduces Rotenone Effect on Stallion Sperm–Zona Pellucida Heterologous Binding

Field fertility of liquid stored and cryopreserved flow cytometrically sex‐sorted stallion sperm

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