Sperm capacitation pretreatment positively impacts bovine intracytoplasmic sperm injection

Águila, Luis; Zambrano, Fabiola; Arias, María Elena; Felmer, Ricardo

doi:10.1002/mrd.22834

Cited by 20 publications

(12 citation statements)

References 99 publications

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“…Additionally, we also observed a significant increase in tyrosine phosphorylation levels under these conditions both in Whitten's and HTF media. Merocyanine 540 has been previously used in boar (Harrison & Miller, ), dog (Steckler et al, ) and bovine sperm (Aguila et al, ) as an indicator of plasma membrane destabilization, because it binds to the segments of the plasma membrane that are in a higher state of lipid disorder (Langner & Hui, ; Leahy & Gadella, ; Williamson et al, ), indicating that changes in the membrane architecture related to sperm capacitation have initiated. These changes apparently result from cholesterol efflux supporting the addition of cholesterol acceptors such as BSA, which are normally included in the capacitation media.…”

Section: Discussionmentioning

confidence: 99%

“…Membrane fluidity, as an early capacitation event, was evaluated using the merocyanine assay (MC540, M6390). MC540 is a hydrophobic dye that stains cell membranes more intensely if their lipid components are in a higher state of disorder (Rathi et al, ) and has been used in dog (Steckler, Stout, Durandt, & Nothling, ), bovine (Aguila, Zambrano, Arias, & Felmer, ) and equine sperm (Ortgies et al, ; Rathi et al, ). Briefly, 5 μl of MC540 (270 μM stock solution) and 2 μl of SYTOX green (20 μM stock solution, Molecular Probes, Eugene, OR, USA) were added to the sperm suspension (2 × 10 6 sperm/ml) and incubated for 30 min at 38.5°C in air and darkness ( n = 3 replicates for each stallion).…”

Section: Methodsmentioning

confidence: 99%

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Effect of human tubal fluid medium and hyperactivation inducers on stallion sperm capacitation and hyperactivation

Arroyo-Salvo

Sanhueza

Fuentes

et al. 2018

Reprod Domestic Animals

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Conventional in vitro fertilization has not yet been implemented in the equine species. One of the main reasons has been the inability to develop a culture medium and incubation conditions supporting high levels of stallion sperm capacitation and hyperactivation in vitro. Although different culture media have been used for this purpose, human tubal fluid (HTF) medium, widely used in the manipulation of human and mice gametes, has not been reported so far in stallion sperm culture. The first part of this study aimed to compare HTF and Whitten's media on different stallion sperm quality and capacitation variables. Additionally, the effect of procaine, aminopyridine and caffeine in both media was evaluated on sperm motility parameters at different incubation times. Integrity and destabilization of the plasma membrane were evaluated by merocyanine 540/SYTOX Green (MC540), mitochondrial membrane potential (∆Ψm) using tetramethylrhodamine methyl ester perchlorate (TMRM), acrosome membrane integrity by PNA/FITC and tyrosine phosphorylation by P-tyrosine mouse mAb conjugated to Alexa Fluor® by flow cytometry. Motility parameters were evaluated using the integrated semen analysis system (ISAS®). We found no differences between Whitten's and HTF media and incubation time in terms of sperm viability, uninduced acrosome membrane damage or mitochondrial membrane potential at 30- and 120-min incubation. Membrane fluidity (MC540) increased in both media at 30- and 120-min incubation compared to noncapacitating conditions. Similarly, tyrosine phosphorylation increased in both media in capacitating conditions at 2- and 4-hr incubation compared to noncapacitating conditions. Although procaine showed the best result in terms of sperm hyperactivated motility in both media, aminopyridine also showed parameters consistent with the hyperactivation including an increase in curvilinear velocity and decrease in straightness. In conclusion, HTF medium and aminopyridine equally support capacitation-related parameters in stallion sperm.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Effect of human tubal fluid medium and hyperactivation inducers on stallion sperm capacitation and hyperactivation

Arroyo-Salvo

Sanhueza

Fuentes

et al. 2018

Reprod Domestic Animals

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“…Hyperactivation and protein tyrosine phosphorylation rises are the best characterized features of sperm capacitation, so advances in the understanding of in vitro capacitation after SU and DGC may help to improve the efficacy and success in ART. Indeed, it has been shown that bovine intracytoplasmic sperm injections using spermatozoa recovered by DGC and pre-incubated under capacitating conditions promote pronucleus formation rate and blastocyst rate [ 28 ].…”

Section: Discussionmentioning

confidence: 99%

Effects of Semen Processing on Sperm Function: Differences between Swim-Up and Density Gradient Centrifugation

Hernández-Silva

López-Torres

Rosas³

et al. 2021

World J Mens Health

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Andrology research has evolved notoriously in the latest years, particularly since male factor contribution to couple infertility has been undoubtedly demonstrated. However, sperm function investigations results are sometimes contradictory, probably as a result of the use of different sperm processing techniques. In this work, we underwent a systematic functional comparison of human sperm samples simultaneously processed by swim-up and density gradient centrifugation, which are the preferred sperm processing methods used in basic and clinical laboratories. Materials and Methods: Materials and Methods: To compare functional characteristics of sperm isolated by swim-up and density gradient centrifugation followed by incubation at different times under capacitating conditions. Results: Results: Semen samples processed in parallel by these two procedures resulted in sperm preparations with significant differences in redox state, spontaneous intracellular calcium oscillations, hyperactivation, protein tyrosine phosphorylation, and acrosome reaction responsivity to calcium ionophore. Such differences showed time-dependent specific patterns for spontaneous intracellular calcium oscillations, hyperactivation and protein tyrosine phosphorylation. Sperm retrieved by density gradient centrifugation showed more hyperactivation and tyrosine phosphorylation than swim-up sperm, suggesting a higher degree of capacitation. Conclusions: Conclusions: Our results account for functional differences observed in spermatozoa processed with these two methods and therefore may contribute to a better interpretation of outcomes obtained in different laboratories as well as to improve experimental designs aimed to study sperm physiology and fertility potential.

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“…The difficulty in remodeling the male chromatin following sperm injection is not unique to the bovine, as earlier studies noted impaired and asynchronous chromatin remodeling following ICSI in the mouse (Shoukir et al 1998, Ajduk et al 2006), pig (Kren et al 2003, Lee et al 2003, Nakai et al 2011) and humans (Dozortsev et al 1994, Sousa & Tesarik 1994, Flaherty et al 1995). Several common factors have been suggested to hinder the remodeling of injected sperm, such as the lack of capacitation and acrosome reaction (Sekhavati et al 2012, Zambrano et al 2016, Aguila et al 2017). Nevertheless, unique features of the bovine spermatozoa may further complicate PN formation in this species.…”

Section: Discussionmentioning

confidence: 99%

Defective sperm head decondensation undermines the success of ICSI in the bovine

et al. 2017

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The efficiency of intracytoplasmic sperm injection (ICSI) in the bovine is low compared to other species. It is unknown whether defective oocyte activation and/or sperm head decondensation limit the success of this technique in this species. To elucidate where the main obstacle lies, we used homologous and heterologous ICSI and parthenogenetic activation procedures. We also evaluated whether maturation negatively impacted the early stages of activation after ICSI. Here we showed that injected bovine sperm are resistant to nuclear decondensation by bovine oocytes and this is only partly overcome by exogenous activation. Remarkably, when we used heterologous ICSI,-matured mouse eggs were capable of mounting calcium oscillations and displaying normal PN formation following injection of bovine sperm, although -matured mouse oocytes were unable to do so. Together, our data demonstrate that bovine sperm are especially resistant to nuclear decondensation by-matured oocytes and this deficiency cannot be simply overcome by exogenous activation protocols, even by inducing physiological calcium oscillations. Therefore, the inability of a suboptimal ooplasmic environment to induce sperm head decondensation limits the success of ICSI in the bovine. Studies aimed to improve the cytoplasmic milieu of -matured oocytes and to replicate the molecular changes associated with capacitation and acrosome reaction will deepen our understanding of the mechanism of fertilization and improve the success of ICSI in this species.

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Sperm capacitation pretreatment positively impacts bovine intracytoplasmic sperm injection

Cited by 20 publications

References 99 publications

Effect of human tubal fluid medium and hyperactivation inducers on stallion sperm capacitation and hyperactivation

Effect of human tubal fluid medium and hyperactivation inducers on stallion sperm capacitation and hyperactivation

Effects of Semen Processing on Sperm Function: Differences between Swim-Up and Density Gradient Centrifugation

Defective sperm head decondensation undermines the success of ICSI in the bovine

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