Liquid storage of equine semen: Assessing the effect of d-penicillamine on longevity of ejaculated and epididymal stallion sperm

Brogan, P.T.; Beitsma, M.; Henning, Heiko; Gadella, B.M.; Stout, T.A.E.

doi:10.1016/j.anireprosci.2015.06.011

Cited by 19 publications

(9 citation statements)

References 47 publications

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“…Previous studies revealed that freezing and thawing-induced apoptosis is inversely correlated with spermatozoa motility and plasma membrane integrity during cryopreservation [15, 19, 22]. Moreover, for liquid storage, the increase in apoptosis exhibiting a significant relationship with the decrease in spermatozoa quality was reported in many species, such as the boar, bull, and stallion [6–8, 35]. In the current study, the occurrence of apoptosis in liquid-stored goat spermatozoa was detected both by TUNEL staining and Annexin-V-FITC/PI staining, which are considered as routine and effective methods to validate spermatozoa undergoing apoptosis and have been utilized in many similar studies [36, 37].…”

Section: Discussionmentioning

confidence: 99%

Mechanisms of ROS-induced mitochondria-dependent apoptosis underlying liquid storage of goat spermatozoa

Liu

Han

Zhou

et al. 2019

Aging

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Liquid storage of spermatozoa is important for artificial insemination and herd genetic breeding. However, the extended time of storage inducing the rapid decline in spermatozoa quality limits the development of this technology. The molecular mechanisms underlying liquid storage of spermatozoa remain largely unexplored. In this study, the effects of liquid storage on functional quality of spermatozoa were assessed in goat (Capra hircus). The time-dependent decline in spermatozoa motility showed a strong correlation with the significant increase in apoptosis. Moreover, apoptosis-related ultrastructural changes were observed, especially the defects in mitochondria. A significant decrease in mitochondrial membrane potential and changes in the expression of mitochondrial apoptosis-related proteins indicated mitochondrial dysfunction and mitochondrial apoptotic pathway activation. Notably, the abnormally high level of reactive oxygen species (ROS) caused by liquid storage resulted in oxidative damage to mitochondria and accelerated mitochondria-dependent apoptosis, as demonstrated by the addition of ROS scavenger N-acetylcysteine. Furthermore, critical differentially expressed proteins involved in mitochondria-dependent apoptosis and antioxidant defense were identified and profiled by quantitative proteomic analysis, facilitating the understanding of molecular regulation of ROS-induced mitochondria-dependent apoptosis. These outcomes provide insights into the mechanisms underlying liquid storage of goat spermatozoa and enhance the progress of semen storage technology.

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Section: Discussionmentioning

confidence: 99%

Mechanisms of ROS-induced mitochondria-dependent apoptosis underlying liquid storage of goat spermatozoa

Liu

Han

Zhou

et al. 2019

Aging

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“…In fact, sperm cryopreservation and sperm liquid storage by short-term refrigeration at +4 °C lead to a significant decline in sperm quality [ 10 ]. In particular, during sperm liquid storage, we observe a deterioration of sperm functions also followed by DNA damage [ 11 ].…”

Section: Introductionmentioning

confidence: 99%

REAC technology as optimizer of stallion spermatozoa liquid storage

Berlinguer

Pasciu

Succu

et al. 2017

Reprod Biol Endocrinol

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BackgroundREAC technology (acronym for Radio Electric Asymmetric Conveyor) is a technology platform for neuro and bio modulation. It has already proven to optimize the ions fluxes at the molecular level and the molecular mechanisms driving cellular asymmetry and polarization.MethodsThis study was designed to verify whether this technology could extend spermatozoa life-span during liquid storage, while preserving their functions, DNA integrity and oxidative status. At 0, 24, 48, and 72 h. of storage at 4 °C, a battery of analyses was performed to assess spermatozoa viability, motility parameters, acrosome status, and DNA integrity during REAC treatment. Spermatozoa oxidative status was assessed by determining lipid peroxidation, the activity of superoxide dismutase (SOD), and the total antioxidant capacity.ResultsDuring liquid storage REAC treated spermatozoa, while not showing an increased viability nor motility compared to untreated ones, had a higher acrosome (p > 0.001) and DNA integrity (p > 0.01). Moreover, the analysis of the oxidative status indicated that the mean activity of the intracellular superoxide dismutase (SOD) was significantly higher in REAC treated spermatozoa compared to untreated controls (p < 0.05), while the intracellular concentration of malondialdehyde (MDA), an end product of lipid peroxidation, at the end of the REAC treatment was higher in untreated controls (p > 0.05). The REAC efficacy on spermatozoa oxidative status was also evidenced by the higher trolox equivalent antioxidant capacity (TEAC) found in both the cellular extract (p < 0.05) and the storage media of REAC treated spermatozoa compared to untreated controls (p < 0.0001).ConclusionThe present study demonstrated that REAC treatment during liquid storage preserves spermatozoa acrosome membrane and DNA integrity, likely due to the enhancement of sperm antioxidant defenses. These results open new perspective about the extending of spermatozoa functions in vitro and the clinical management of male infertility.Electronic supplementary materialThe online version of this article (doi:10.1186/s12958-017-0229-6) contains supplementary material, which is available to authorized users.

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“…Sample concentration was measured with a Bürker Türk haemocytometer and ejaculates were diluted in INRA 96 ® to a concentration of 30 × 10 6 spermatozoa/mL. Motility was checked objectively using a computer-assisted semen analysis (CASA) system (SpermVision 3.5, Minitüb, Tiefenbach, Germany) as described by Brogan et al (2015) . Only samples with greater than or equal to 70% (total) motile sperm in the diluted semen were used for experiments.…”

Section: Methodsmentioning

confidence: 99%

Bicarbonate-Stimulated Membrane Reorganization in Stallion Spermatozoa

Maitan

Bromfield

Hoogendijk

et al. 2021

Front. Cell Dev. Biol.

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Classical in vitro fertilization (IVF) is still poorly successful in horses. This lack of success is thought to be due primarily to inadequate capacitation of stallion spermatozoa under in vitro conditions. In species in which IVF is successful, bicarbonate, calcium, and albumin are considered the key components that enable a gradual reorganization of the sperm plasma membrane that allows the spermatozoa to undergo an acrosome reaction and fertilize the oocyte. The aim of this work was to comprehensively examine contributors to stallion sperm capacitation by investigating bicarbonate-induced membrane remodelling steps, and elucidating the contribution of cAMP signalling to these events. In the presence of capacitating media containing bicarbonate, a significant increase in plasma membrane fluidity was readily detected using merocyanine 540 staining in the majority of viable spermatozoa within 15 min of bicarbonate exposure. Specific inhibition of soluble adenylyl cyclase (sAC) in the presence of bicarbonate by LRE1 significantly reduced the number of viable sperm with high membrane fluidity. This suggests a vital role for sAC-mediated cAMP production in the regulation of membrane fluidity. Cryo-electron tomography of viable cells with high membrane fluidity revealed a range of membrane remodelling intermediates, including destabilized membranes and zones with close apposition of the plasma membrane and the outer acrosomal membrane. However, lipidomic analysis of equivalent viable spermatozoa with high membrane fluidity demonstrated that this phenomenon was neither accompanied by a gross change in the phospholipid composition of stallion sperm membranes nor detectable sterol efflux (p > 0.05). After an early increase in membrane fluidity, a significant and cAMP-dependent increase in viable sperm with phosphatidylserine (PS), but not phosphatidylethanolamine (PE) exposure was noted. While the events observed partly resemble findings from the in vitro capacitation of sperm from other mammalian species, the lack of cholesterol removal appears to be an equine-specific phenomenon. This research will assist in the development of a defined medium for the capacitation of stallion sperm and will facilitate progress toward a functional IVF protocol for horse gametes.

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Liquid storage of equine semen: Assessing the effect of d-penicillamine on longevity of ejaculated and epididymal stallion sperm

Cited by 19 publications

References 47 publications

Mechanisms of ROS-induced mitochondria-dependent apoptosis underlying liquid storage of goat spermatozoa

Mechanisms of ROS-induced mitochondria-dependent apoptosis underlying liquid storage of goat spermatozoa

REAC technology as optimizer of stallion spermatozoa liquid storage

Bicarbonate-Stimulated Membrane Reorganization in Stallion Spermatozoa

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