Estrous sheep serum as a potent agent for ovine IVF: Effect on cholesterol efflux from spermatozoa and the acrosome reaction

Huneau, D.; Crozet, Nicole; Ahmed-Ali, Michèle

doi:10.1016/0093-691x(94)90123-z

Cited by 32 publications

(20 citation statements)

References 24 publications

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“…In our study, when SOF was supplemented with BSA-heparin-hypotaurine (SOF BSA ), the proportion of spermatozoa displaying high membrane fluidity was significantly higher at the beginning of incubation, when compared to all other media tested with the exception of SOF ESS which yielded similar results. It was previously shown that ESS supported cholesterol efflux from the plasma membrane of ram spermatozoa [32]. Moreover, on the basis of LIN and ALH results at 60 minutes of incubation, spermatozoa treated with both BSA or ESS developed hyperactivated motility, one of the hallmarks of sperm capacitation [3].…”

Section: Discussionmentioning

confidence: 98%

“…Despite advances in knowledge regarding additives that may support mammalian sperm capacitation in vitro, in small ruminants, incubation of sperm in the presence of estrous sheep serum (ESS) is still required to achieve optimal rates of IVF [21,[31][32][33]. However, a main drawback of using ESS is that it is not easily attainable given that it requires extraction and purification from blood samples and, therefore, its composition can be variable from batch to batch.…”

Section: Introductionmentioning

confidence: 98%

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Effect of different media additives on capacitation of frozen–thawed ram spermatozoa as a potential replacement for estrous sheep serum

García-Álvarez¹,

Maroto-Morales²,

Jiménez-Rabadán

et al. 2015

Theriogenology

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Capacitation is a key process through which spermatozoa acquire their fertilizing ability. This event is required for the successful application of assisted reproductive technologies such as IVF. The aim of the present study was to investigate the effect of using a synthetic oviductal fluid medium supplemented with either heparin-hypotaurine alone, in combination with progesterone (P4), 17β-estradiol (E2), or BSA, or just β-cyclodextrin, in replacement for estrous sheep serum (ESS) for ram sperm capacitation. After incubation in the corresponding media for 15 (time 0) or 60 minutes, sperm function was evaluated by computerized sperm motility analysis and flow cytometry (plasma membrane status and fluidity). Treatments rendering the best results in regards to sperm function parameters related to capacitation were used for an IVF test. Herein, neither heparin-hypotaurine (alone), or in combination with P4, or E2, nor β-cyclodextrin induced capacitation-related changes in frozen-thawed ram spermatozoa. Only the medium supplemented with heparin-hypotaurine-BSA was able to induce changes compatible with in vitro capacitation relating to sperm motility pattern and plasma membrane fluidity, comparable to those in ESS-containing medium. Both media yielded sperm parameter values that differed (P < 0.05) from those obtained in the rest of the media tested. However, after the IVF trial, BSA was unable to support cleavage rates (21.80%) comparable to those obtained with ESS (52.60%; P < 0.05). We conclude that heparin-hypotaurine, P4, E2, β-cyclodextrin, or BSA is not suitable for replacing ESS in capacitation and fertilization media for ram spermatozoa.

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Section: Discussionmentioning

confidence: 98%

Section: Introductionmentioning

confidence: 98%

Effect of different media additives on capacitation of frozen–thawed ram spermatozoa as a potential replacement for estrous sheep serum

García-Álvarez¹,

Maroto-Morales²,

Jiménez-Rabadán

et al. 2015

Theriogenology

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“…TALP, SOF) do not stimulate protein tyrosine phosphorylation and cholesterol efflux of ram spermatozoa (Huneau et al 1994, Li et al 2006, Colas et al 2008, Leahy & Gadella 2015. Since sheep serum has been previously reported to cause sperm agglutination (Senger & Saacke 1976, Lindahl & Sjö blom 1981, Senger et al 1981, and SOF supplemented with serum (2-20%) is the most commonly used sheep IVF fertilising medium, we tested if PEN could prevent agglutination in SOF medium containing 2.5% or 10% sheep serum.…”

Section: Discussionmentioning

confidence: 99%

“…Ram spermatozoa have proven to be difficult to capacitate in vitro and require supplementation with both dibutyryl-cAMP (db-cAMP) and phosphodiesterase (PDE) inhibitors to stimulate the cAMP pathway and achieve maximal tyrosine phosphorylation of sperm proteins (Colas et al 2008). Ram spermatozoa also show a unique requirement for sheep serum to be present in the fertilising medium for IVF success (Huneau et al 1994, Li et al 2006 as TALP alone does not stimulate cholesterol efflux in this species (Huneau et al 1994, Colas et al 2008, Leahy & Gadella 2015. Another example is the addition of the highly negatively charged glycosaminoglycan heparin to bovine IVF protocols (Parrish et al 1988).…”

Section: Introductionmentioning

confidence: 99%

Penicillamine prevents ram sperm agglutination in media that support capacitation

Leahy¹,

Rickard²,

Aitken³

et al. 2016

Reproduction

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Ram spermatozoa are difficult to capacitate in vitro. Here we describe a further complication, the unreported phenomenon of headto-head agglutination of ram spermatozoa following dilution in the capacitation medium Tyrodes plus albumin, lactate and pyruvate (TALP). Sperm agglutination is immediate, specific and persistent and is not associated with a loss of motility. Agglutination impedes in vitro sperm handling and analysis. So the objectives of this study were to investigate the cause of sperm agglutination and potential agents which may reduce agglutination. The percentage of non-agglutinated, motile spermatozoa increased when bicarbonate was omitted from complete TALP suggesting that bicarbonate ions stimulate the agglutination process. D-penicillamine (PEN), a nucleophilic thiol, was highly effective at reducing agglutination. The inclusion of 250 mM PEN in TALP reduced the incidence of motile, agglutinated spermatozoa from 76.7G2.7% to 2.8G1.4%. It was then assessed if PEN (1 mM) could be included in existing ram sperm capacitation protocols (TALP C1 mM dibutyryl cAMP, caffeine and theophylline) to produce spermatozoa that were simultaneously capacitated and non-agglutinated. This protocol resulted in a sperm population which displayed high levels of tyrosine phosphorylated proteins and lipid disordered membranes (merocyanine-540) while remaining motile, viable, acrosome-intact and non-agglutinated. In summary, PEN (1 mM) can be included in ram sperm capacitation protocols to reduce sperm agglutination and allow for the in vitro assessment of ram sperm capacitation.

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“…Female biological fluids have also been used for cholesterol efflux from sperm. Huneau et al (60) used 20% estrous sheep serum in an incubation medium for cholesterol efflux and reported higher cholesterol efflux (60% vs. 14%) and fertility (85% vs. 0.7%) in the treated ram sperm compared to the control. In another study, Langlais et al (19) incubated human spermatozoa in a medium supplemented with 5% human female serum or follicular fluid.…”

Section: Miscellaneous Agentsmentioning

confidence: 99%

Cholesterol efflux from sperm: approaches and applications

Naseer¹,

Ahmad²,

Aksoy³

2014

Turk J Vet Anim Sci

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Following ejaculation, sperm appear morphologically mature, but functionally they are inactive. To attain functional capability or capacitation in vivo, sperm must reside in the female reproductive tract for a certain period of time before fertilization. In the female reproductive tract, cholesterol efflux from the sperm plasma membrane is considered vital for the initiation of capacitation. A number of in vitro studies have used biological and nonphysiological means to understand the phenomenon of cholesterol efflux and capacitation. In this review, we present a short overview of the role of cholesterol efflux in regulating sperm capacitation using various agents. This review provides insight to researchers designing new procedures for sperm preparation to be used in assisted reproductive technology.

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Estrous sheep serum as a potent agent for ovine IVF: Effect on cholesterol efflux from spermatozoa and the acrosome reaction

Cited by 32 publications

References 24 publications

Effect of different media additives on capacitation of frozen–thawed ram spermatozoa as a potential replacement for estrous sheep serum

Effect of different media additives on capacitation of frozen–thawed ram spermatozoa as a potential replacement for estrous sheep serum

Penicillamine prevents ram sperm agglutination in media that support capacitation

Cholesterol efflux from sperm: approaches and applications

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