Lycopene and resveratrol improve post-thaw bull sperm parameters: sperm motility, mitochondrial activity and DNA integrity

The aim of this study was to determine the effects of trehalose and cysteine on sperm motility, viability, mitochondrial activity and acrosome integrity during liquid storage of Merino ram semen. Ejaculates were collected using artificial vaginas from five Merino rams, microscopically evaluated and pooled at 37 °C. The pooled semen samples were diluted in a Tris-based extender, including cysteine (2 mM and 4 mM), trehalose (10 mM and 25 mM) and no antioxidant (control). Diluted semen samples were kept in tubes and cooled from 37 to 5 °C in a cold cabinet, and maintained at 5 °C. Cooled samples were evaluated for sperm motility, viability, mitochondrial activity and acrosome integrity at 0, 24, 48, 72 and 96 h. Extender supplemented with trehalose (10 and 25 mM) and cysteine (2 and 4 mM) led to higher motility in comparison to the control at 24, 48, 72 and 96 h of liquid storage (P < 0.05). Trehalose at the doses of 10 mM, 25 mM and 2 mM cysteine led to higher viability between 24–48–72 h and at 96 h of liquid storage (P < 0.05). Further, 4 mM of cysteine improved sperm viability rates at 24 and 48 h of storage compared to the control group (P < 0.05), and resulted in improved acrosome integrity rates compared to the control group at 72 and 96 h of storage (P < 0.05). Extender supplemented with 10 and 25 mM trehalose at 24 and 72 h and 4 mM cysteine at 24 and 96 h of storage led to higher sperm mitochondrial activity rates when compared to the control group (P < 0.05). In conclusion, the findings of this study show that trehalose and cysteine provided significant protection to ram sperm parameters during liquid storage.

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Epicatechin Provides Antioxidant Protection to Bovine Spermatozoa Subjected to Induced Oxidative Stress

Molecules ◽

10.3390/molecules24183226 ◽

2019 ◽

Vol 24 (18) ◽

pp. 3226 ◽

Cited By ~ 5

Author(s):

Eva Tvrda ◽

Peter Straka ◽

Drahomir Galbavy ◽

Peter Ivanic

Keyword(s):

Oxidative Damage ◽

Sperm Motility ◽

Reproductive Technologies ◽

Protein Carbonyls ◽

Mitochondrial Activity ◽

Dna Integrity ◽

Ros Production ◽

Ros Scavenging ◽

Bovine Spermatozoa ◽

Dispersion Test

Epicatechin (EPI) is a natural flavonoid with antibacterial, anti-inflammatory and anti-cancer properties. Furthermore, the molecule exhibits powerful reactive oxygen species (ROS) scavenging and metal-chelating properties. In this study, we assessed the efficiency of EPI to reverse ROS-mediated alterations to the motility, viability, DNA integrity and oxidative profile of bovine spermatozoa. For the first experiment, spermatozoa were washed out of fresh semen and exposed to 12.5 μmol/L EPI, 25 μmol/L EPI, 50 μmol/L EPI and 100 μmol/L EPI in the presence of ferrous ascorbate (FeAA) during a 6 h in vitro culture. For the second experiment, the ejaculates were split into aliquots and cryopreserved with a commercial semen extender supplemented with 12.5 μmol/L EPI, 25 μmol/L EPI, 50 μmol/L EPI, 100 μmol/L EPI or containing no supplement. Sperm motility was assessed using the computer-aided sperm analysis and the cell viability was studied with the metabolic activity test. ROS production was quantified using luminometry, and DNA fragmentation was evaluated using the chromatin dispersion test. Cell lysates were prepared at the end of the culture in order to assess the concentration of protein carbonyls and malondialdehyde. Exposure to FeAA led to a significantly reduced sperm motility (p < 0.001), mitochondrial activity (p < 0.001), but increased the generation of ROS (p < 0.001), as well as oxidative damage to proteins (p < 0.001), DNA (p < 0.001) and lipids (p < 0.001). EPI supplementation, particularly at a concentration range of 50–100 μmol/L, resulted in higher preservation of the spermatozoa vitality (p < 0.001). Furthermore, 50–100 μmol/L EPI were significantly effective in the prevention of oxidative damage to sperm proteins (p < 0.001), lipids (p < 0.001) and DNA (p < 0.01 in relation to 50 μmol/L EPI; p < 0.001 with respect to 100 μmol/L EPI). In the case of the cryopreserved spermatozoa, the administration of 50–100 μmol/L EPI resulted in higher sperm motility (p < 0.001) and mitochondrial activity (p < 0.001). ROS production, the number of protein carbonyls, lipid peroxidation as well as oxidative DNA damage were found to be significantly decreased particularly in samples cryopreserved in the presence of 100 μmol/L EPI (p < 0.001). Our results suggest that EPI could behave as an effective antioxidant which may prevent oxidative insults to spermatozoa, and thus, preserve their vitality and functionality. Nevertheless, its potential to achieve higher fertilization rates in reproductive technologies needs to be validated.

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Red-Light Irradiation of Horse Spermatozoa Increases Mitochondrial Activity and Motility through Changes in the Motile Sperm Subpopulation Structure

Biology ◽

10.3390/biology9090254 ◽

2020 ◽

Vol 9 (9) ◽

pp. 254 ◽

Cited By ~ 3

Author(s):

Jaime Catalán ◽

Marion Papas ◽

Sabrina Gacem ◽

Yentel Mateo-Otero ◽

Joan E. Rodríguez-Gil ◽

...

Keyword(s):

Sperm Motility ◽

Consumption Rate ◽

Red Light ◽

Light Irradiation ◽

Mitochondrial Activity ◽

Dna Integrity ◽

O2 Consumption ◽

Sperm Viability ◽

Intracellular Atp ◽

Atp Levels

Previous studies in other mammalian species have shown that stimulation of semen with red-light increases sperm motility, mitochondrial activity, and fertilizing capacity. This study sought to determine whether red-light stimulation using a light emitting diode (LED) at 620–630 nm affects sperm motility and structure of motile subpopulations, sperm viability, mitochondrial activity, intracellular ATP levels, rate of O2 consumption and DNA integrity of horse spermatozoa. For this purpose, nine ejaculates were collected from nine different adult stallions. Upon collection, semen was diluted in Kenney extender, analyzed, its concentration was adjusted, and finally it was stimulated with red-light. In all cases, semen was packaged in 0.5-mL transparent straws, which were randomly divided into controls and 19 light-stimulation treatments; 6 consisted of a single exposure to red-light, and the other 13 involved irradiation with intervals of irradiation and darkness (light-dark-light). After irradiation, sperm motility was assessed using a Computerized Semen Analysis System (CASA). Flow cytometry was used to evaluate sperm viability, mitochondrial membrane potential and DNA fragmentation. Intracellular levels of ATP and O2 consumption rate were also determined. Specific red-light patterns were found to modify kinetics parameters (patterns: 4, 2-2-2, 3-3-3, 4-4-4, 5-1-5, and 5-5-5 min), the structure of motile sperm subpopulations (patterns: 2, 2-2-2, 3-3-3, and 4-1-4 min), mitochondrial membrane potential (patterns: 4, 3-3-3, 4-4-4, 5-1-5, 5-5-5, 15-5-15, and 15-15-15 min), intracellular ATP levels and the rate of O2 consumption (pattern: 4 min), without affecting sperm viability or DNA integrity. Since the increase in some kinematic parameters was concomitant with that of mitochondrial activity, intracellular ATP levels and O2 consumption rate, we suggest that the positive effect of light-irradiation on sperm motility is related to its impact upon mitochondrial activity. In conclusion, this study shows that red LED light stimulates motility and mitochondrial activity of horse sperm. Additional research is needed to address the impact of red-light irradiation on fertilizing ability and the mechanisms through which light exerts its effects.

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26 SPERM REPRODUCTIVE POTENTIAL OF FETAL FIBROBLAST CELLS ON A NUCLEAR TRANSFER - CLONED BULL AND ON AN ORIGINAL PATERNAL BULL: A COMPARATIVE ANALYSIS

Reproduction Fertility and Development ◽

10.1071/rdv19n1ab26 ◽

2007 ◽

Vol 19 (1) ◽

pp. 131

Author(s):

S.-H. Bae ◽

D.-H. Kim ◽

S.-W. Kim ◽

G.-S. Im ◽

M.-J. Lee ◽

...

Keyword(s):

Sperm Motility ◽

Treated Group ◽

Flow Cytometer ◽

Mitochondrial Activity ◽

Computer Assisted ◽

Fibroblast Cells ◽

Sperm Viability ◽

Fetal Fibroblast ◽

Reproductive Ability ◽

Bull Sperm

Recently, cloning technology has produced newborn calves from fetal fibroblast cells and somatic cells. These cloned calves appear to be normal and healthy in terms of reproductive ability. Therefore, we undertook this study to compare the reproductive ability of cloned sperm from the clones themselves and sperm obtained from the frozen semen of the paternal bull (KPN-178). Two cloned bulls from fetal fibroblast cell nuclear transfers (Korean HanWoo Clone-38, 34 months old, and Clone-39, 30 months old) were selected. Sperm from the cloned groups (C-38 and C-39), cloned-38 fresh sperm, and sperm from the paternal bull (KPN-178) were analyzed for the following sperm reproductive characteristics: sperm motility, by computer-assisted sperm analysis (CASA); sperm mitochondrial activity, by flow cytometer after mitotracker staining; and sperm viability, by flow cytometer after staining with live/dead sperm viability kits, both PI and SYBR. Sperm motility was evaluated with Percoll-treated or non-Percoll-treated frozen-thawed spermatozoa. In the Percoll-treated group, total motility of the paternal bull sperm (KPN-178: 75.14%) was significantly lower than that of cloned sperm (C-38: 90.42%; C-39: 87.62%: C-38 fresh: 93.97%; P < 0.05). The non-Percoll-treated group showed different results compared to the Percoll results (P < 0.05), i.e. the total motility of C-38 (92.47%) and C-38 fresh (92.47%) sperm was significantly higher than that of C-39 (79.52%) and KPN-178 (78.41%) sperm. The mitochondrial activity staining rate of KPN-178 (58.22%) sperm was low compared to that of C-38 (88.65%), C-39 (89.45%), C-38 fresh (88.66%) sperm. The sperm viability tests showed similar results (P < 0.05) for all sperm groups: C-38 (PI: 42.53% and SYBR: 46.28%), C-39 (PI: 39.03% and SYBR: 66.18%), C-38 fresh (PI: 34.32% and SYBR: 57.10%), and KPN-178 (PI: 14.43% and SYBR: 47.86%). However, the PI staining of KPN-178 was lower than that of the other sperm groups. Next, in vitro fertilization (IVF) was investigated to compare the reproductive ability of C-38, C-39, KPN-178, and Control (normal bull) sperm. The results, development to the 2-cell stage (157/228, 68.90%; 85/116, 73.28%; 78/114, 68.42%; 61/79, 77.21%, respectively) and blastocyst rates (41/228, 18.0%; 35/116, 30.17%; 26/114, 22.81%; 20/79, 25.32%, respectively) were evaluated (P < 0.05). The IVF rates were not different among cloned bull, paternal bull, and normal bull sperm. A cloned female bovine was successfully impregnated by cloned bull sperm (C-38) using artificial insemination. The calf resulting from this prenancy showed no signs of phenotypical abnormalities. These results suggest that the physiology of cloned bulls and the quality of semen from cloned bulls show no evidence of any deleterious effects.

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Effects of arginine and trehalose on post-thawed bovine sperm quality

Acta Veterinaria Hungarica ◽

10.1556/004.2017.040 ◽

2017 ◽

Vol 65 (3) ◽

pp. 429-439 ◽

Cited By ~ 2

Author(s):

Caner Öztürk ◽

Şükrü Güngör ◽

Mehmet Bozkurt Ataman ◽

Mustafa Numan Bucak ◽

Nuri Başpinar ◽

...

Keyword(s):

Sperm Quality ◽

Protective Role ◽

Mitochondrial Activity ◽

Dna Integrity ◽

Bovine Sperm ◽

Sperm Dna ◽

Bull Sperm ◽

Semen Extender ◽

And Oxidative Stress

The present study was conducted to examine the protective role of arginine and trehalose on post-thaw bull sperm and oxidative stress parameters. Five ejaculates for each bull were used in the study. Each ejaculate, split into three equal aliquots and diluted at 37 °C with base extenders containing 2 mM arginine, 25 mM trehalose and no antioxidant (control) was cooled to 5 °C and then frozen. Frozen straws were thawed in a water bath for evaluation. Supplementation of the semen extender with arginine decreased the percentages of post-thawed subjective motility (29 ± 8.21%), CASA motility (12.2 ± 5.69%) and progressive motility (3.52 ± 2.13%), compared with the controls (43 ± 2.73%, 55.4 ± 6.78% and 33.48 ± 4.14%, respectively, P < 0.05). Supplementation of the semen extender with trehalose produced a higher mitochondrial activity and sperm viability (36.3 ± 3.99% and 44.1 ± 2.18%) compared with the control (13 ± 8.15 and 31.7 ± 3.94%, respectively, P < 0.05). It was established that trehalose (95.1%) and arginine (92.8%) protect DNA integrity compared to the control (90.4%) (P < 0.05). Trehalose supplementation in semen extenders provided great benefit in terms of viability, mitochondrial activity, and intact sperm DNA on frozen-thawed bull sperm.

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Crocin Improves the Quality of Cryopreserved Goat Semen in Different Breeds

Animals ◽

10.3390/ani10061101 ◽

2020 ◽

Vol 10 (6) ◽

pp. 1101

Author(s):

Valentina Longobardi ◽

Gianluigi Zullo ◽

Alessio Cotticelli ◽

Angela Salzano ◽

Giuseppe Albero ◽

...

Keyword(s):

Dna Fragmentation ◽

Sperm Motility ◽

Membrane Integrity ◽

Egg Yolk ◽

Sperm Parameters ◽

Freezing Process ◽

Control Group ◽

Dna Integrity ◽

Semen Extender ◽

Dose Trial

The effect of crocin in the semen extender before cryopreservation was evaluated on sperm parameters of 20 bucks of five different breeds: Garganica (GA), Jonica (JO), Maltese (MA), Mediterranean Red (MR) and Saanen (SA). Semen samples were centrifuged, to remove seminal plasma, divided in two aliquots and diluted with Tris-egg-yolk-based extender, containing 0 (control group) and 1 mM crocin. Crocin concentration was established after a preliminary dose trial. On fresh and frozen-thawed sperm, motility, viability, morphology, membrane integrity, DNA fragmentation and ROS levels were evaluated. The freezing process led to a decrease (p < 0.05) in all the sperm parameters recorded, confirming the deleterious effect of cryopreservation on goat semen. The most interesting result regarding the inclusion of crocin in the extender before cryopreservation was as follows: Crocin significantly improved (p < 0.05) sperm motility in all breeds, except for Mediterranean Red, compared to the control group. Furthermore, 1 mM crocin reduced percentage of spermatozoa with DNA fragmentation with a marked decrement (p < 0.05) in Garganica and Saanen, as compared to the control group. Finally, intracellular ROS decreased (p < 0.01) in the crocin-treated sperm of all breeds, as compared to the control. In conclusion, supplementation of 1 mM crocin in the extender decreased oxidative stress, improving sperm motility and the DNA integrity of frozen-thawed sperm in different breeds.

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