Comprehensive Flow-Cytometric Quality Assessment of Ram Sperm Intended for Gene Banking Using Standard and Novel Fertility Biomarkers

Vašíček, Jaromír; Baláži, Andrej; Svoradová, Andrea; Vozaf, Jakub; Dujíčková, Linda; Makarevich, Alexander V.; Bauer, Miroslav; Chrenek, Peter

doi:10.3390/ijms23115920

Cited by 20 publications

(12 citation statements)

References 83 publications

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“…Different probes can be used to assess ROS and changes in the oxidative status of ram sperm supplemented with different antioxidants. Some probes are specific and react with a particular anion (e.g., superoxide anion), and others are nonspecific and react with several types of ROS [ 61 ]. Fluorescent probes are employed to evaluate sperm capacitation, acrosome and plasma membrane integrity.…”

Section: Discussionmentioning

confidence: 99%

Effect of In Vitro Addition of Melatonin and Glutathione on Seminal Parameters of Rams in Diluted Semen and after Thawing

Carrico

Barbas

Pimenta

et al. 2023

Veterinary Sciences

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The present study aimed to evaluate the effects of melatonin (MLT), glutathione (GSH), and their combination on ram semen quality after thawing. During eight weekly sessions, semen from three Merino rams was pooled, diluted with an egg-yolk-based semen extender, and divided into four groups: control, 1 mM MLT, 5 mM GSH, and 1 mM MLT + 5 mM GSH. Diluted semen was evaluated before and after the freezing process. The supplementation of diluted semen with GSH at 5 mM had a deleterious effect on total motility progressive (TPM), curvilinear velocity (VCL), straight-line velocity (VSL), average-path velocity (VAP), linearity (LIN), and straightness (STR) and increased slow spermatozoa (%). MLT at 1 mM also had a negative effect on TPM, VSL, and STR in diluted semen. In thawed semen, 1 mM MLT increased the TPM compared with the control group. VSL was lower in the 5 mM GSH group than in the 1 mM MLT group. Additionally, the combination of both antioxidants attenuated the negative effect of 5 mM GSH on TPM, VSL, and BCF. These results indicate that 5 mM GSH impairs or does not improve sperm kinetic parameters in either diluted or thawed semen. They also suggest that MLT combined with GSH plays a protective role against these effects.

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Section: Discussionmentioning

confidence: 99%

Effect of In Vitro Addition of Melatonin and Glutathione on Seminal Parameters of Rams in Diluted Semen and after Thawing

Carrico

Barbas

Pimenta

et al. 2023

Veterinary Sciences

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“…There are different interpretations of the expression “sperm quality”; in this review, it is taken to mean an indirect assessment of the potential ability of a sperm sample to achieve fertilization. Thus, factors such as sperm motility, membrane integrity, acrosome integrity, chromatin integrity, and mitochondrial function, are considered to be indicative of future fertilizing potential [ 13 ]. Other methods, such as “omics” or molecular assays, require even more advanced instrumentation and expertise.…”

Section: Sperm Quality Pre- and Post-cryopreservationmentioning

confidence: 99%

Freezing Stallion Semen—What Do We Need to Focus on for the Future?

Al-Kass,

Morrell

2024

Veterinary Sciences

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Artificial insemination (AI) is used frequently in the breeding of sport horses, apart from Thoroughbreds. Most AIs are carried out with cooled semen rather than frozen semen because of the difficulties in identifying a protocol that is suitable for freezing most ejaculates and the necessity to inseminate close to ovulation because of the short life of the thawed spermatozoa. More widespread use of frozen semen would improve biosecurity, allow greater choice of stallions, and offer more flexibility when managing deliveries of semen to the stud. It would even decrease the amount of antibiotics used in semen extenders, since the volume of frozen semen is smaller than when cooled semen is inseminated. However, there is considerable variability in the cryosurvival of spermatozoa from different stallions, leading to the classification of stallions as good or bad freezers. Improvements could be made at the level of stallion nutrition, the semen collection regimen, the extender, the removal of seminal plasma, and the cooling protocol, among others. Stallion sperm membranes are highly susceptible to lipid peroxidation, but research on antioxidants has failed to identify an additive that would benefit all stallions. In the future, biomarkers for sperm freezability could be used as an aid in identifying suitable ejaculates for cryopreservation.

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“…All chemicals were sourced from ThermoFisher Scientific (USA) unless specified otherwise. Fresh and post-thaw semen samples were incubated with the above-mentioned reagents either following the producer's manuals or as described previously (Vasicek et al 2022).…”

Section: Flow Cytometry Analysesmentioning

confidence: 99%

The effect of ellagic acid on rabbit sperm in vitro parameters after cryopreservation

Grba,

Kuželová,

Makarevich

et al. 2024

Czech J. Anim. Sci.

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Cryopreservation reduces the antioxidant activity of spermatozoa and makes them more sensitive to damage caused by reactive oxygen species (ROS). The addition of antioxidants to the freezing medium could prevent cryo-damage by mitigating the harmful effects of ROS and, thus, protecting the spermatozoa. This study aimed to evaluate the effect of ellagic acid (EA) on the rabbit sperm traits after freezing-thawing. Semen samples collected from New Zealand White rabbit males were cryopreserved in a BotuCrio freezing medium (Nidacon, Sweden) supplemented with different concentrations of ellagic acid (EA at 0, 0.5, 1.5 and 2.5 mM) using the manual slow freezing procedure. After thawing, sperm motility parameters were evaluated by CASA. The parameters of viability (DRAQ7), apoptosis (Yo-Pro-1), acrosome integrity (peanut agglutinin; PNA), intracellular ROS (Cell-ROX) and mitochondrial activity (MitoTracker) were evaluated by flow cytometry. EA added to the freezing medium at all concentrations led to a significant reduction (P < 0.05) in intracellular ROS in frozen-thawed sperm cells. However, this effect was not reflected in motility parameters. Semen supplemented with 1.5 mM EA also yielded a lower proportion of apoptotic cells compared to the control group. In conclusion, EA supplementation of semen extender demonstrated its antioxidative properties protecting spermatozoa against oxidative damage during cryopreservation. Nevertheless, to draw a definitive conclusion regarding the effect of EA on spermatozoa functionality, additional research is necessary.

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Comprehensive Flow-Cytometric Quality Assessment of Ram Sperm Intended for Gene Banking Using Standard and Novel Fertility Biomarkers

Cited by 20 publications

References 83 publications

Effect of In Vitro Addition of Melatonin and Glutathione on Seminal Parameters of Rams in Diluted Semen and after Thawing

Effect of In Vitro Addition of Melatonin and Glutathione on Seminal Parameters of Rams in Diluted Semen and after Thawing

Freezing Stallion Semen—What Do We Need to Focus on for the Future?

The effect of ellagic acid on rabbit sperm in vitro parameters after cryopreservation

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