Antioxidant Effect of a Polyphenol-Rich Murtilla (Ugni molinae Turcz.) Extract and Its Effect on the Regulation of Metabolism in Refrigerated Boar Sperm

The production of reactive oxygen species (ROS) in boar spermatozoa increases in refrigeration; this can have an impact on sperm quality and fertilization capacity. We evaluated the effect of polyphenol-rich aqueous extract of murtilla (Ugni molinae Turcz) on boar sperm stored at 17°C in order to reduce oxidative stress and improve sperm quality in the long term. Five experiments were performed: first, characterization of the polyphenol content from five genotypes of murtilla; second, determination of the genotype with the best antioxidant effect (MT-Ex); third, the antioxidant capacity on O2- and lipid peroxidation; fourth, the influence of MT-Ex on motility, calcium movement, cAMP, and metabolic parameters; and fifth, analysis of long-term refrigeration. The average phenolic content was 344 ppm; gallic acid, catechin, quercetin, myricetin, and kaempferol were detected. All extracts evaluated presented a concentration-dependent antioxidant effect. MT-Ex reduces intracellular O2-/peroxides but low lipid peroxidation. MT-Ex in nonstimulated ROS conditions reduces sperm motility, mitochondrial membrane potential, cAMP, and ATP, but the succinate dehydrogenase activity remained normal; also, we observed a reduction in calcium movement in in vitro sperm capacitation. The long-term analyses showed that MT-Ex improved sperm motility decay and reduced membrane damage and ROS at 168 h. Based on this study, we propose MT-Ex as a supplement in semen extenders.

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Effect of L-carnitine on sperm quality during liquid storage of boar semen

Asian-Australasian Journal of Animal Sciences ◽

10.5713/ajas.19.0455 ◽

2020 ◽

Vol 33 (11) ◽

pp. 1763-1769

Author(s):

Kang Yang ◽

Na Wang ◽

Hai-Tao Guo ◽

Jing-Ran Wang ◽

Huan-Huan Sun ◽

...

Keyword(s):

Sperm Motility ◽

Sperm Quality ◽

Membrane Integrity ◽

Quality Parameters ◽

Atp Content ◽

Developmental Potential ◽

Positive Effects ◽

Liquid Storage ◽

Boar Sperm

Objective: This study was conducted to investigate the effect of L-carnitine on the pig semen characteristics during storage.Methods: Spermatozoa samples were examined for spermatozoa quality and then randomly divided into 5 groups: 0 (control), 12.5, 25, 50, and 100 mM L-carnitine. Sperm motility, plasma membrane integrity and antioxidant parameters (total reactive oxygen species, total antioxidant capacity, and malondialdehyde) were evaluated after 0, 3, 5, and 10 day cooledstorage at 17°C. Moreover, ATP content, mitochondria activity as well as sperm-binding and in vitro fertilizing ability of preserved boar sperm were also investigated.Results: Supplementation with 50 mM L-carnitine could effectively maintain boar sperm quality parameters such as sperm motility and membrane integrity. Besides, we found that L-carnitine had positive effects on boar sperm quality mainly through improving antioxidant capacities and enhancing ATP content and mitochondria activity. Interestingly, by assessing the effect of L-carnitine on sperm fertility and developmental potential, we discovered that the extender containing L-carnitine could improve sperm quality and increase the number of sperms bounding to zona pellucida, without improving in vitro fertility and development potential.Conclusion: These findings suggested that the proper addition of L-carnitine to the semen extender improved boar sperm quality during liquid storage at 17°C.

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Cooling of equine semen at 16°C for 36 hours with addition of different glutathione concentrations

Semina Ciências Agrárias ◽

10.5433/1679-0359.2015v36n6p3699 ◽

2015 ◽

Vol 36 (6) ◽

pp. 3699

Author(s):

Rodrigo Arruda de Oliveira ◽

Marco Antônio De Oliveira Viu ◽

Maria Lúcia Gambarini

Keyword(s):

Lipid Peroxidation ◽

Sperm Motility ◽

Membrane Integrity ◽

Membrane Lipid ◽

Sperm Cells ◽

Sperm Viability ◽

Membrane Lipid Peroxidation ◽

Acrosomal Membrane ◽

In Vitro Effects

Handling equine semen during the refrigeration process reduces sperm viability, and consequently causes membrane lipid peroxidation, among other challenges. The present study aimed to evaluate the in vitro effects of glutathione (control, 1. 0, 1. 5, and 2. 5 mM) on equine semen in a refrigeration protocol of 16ºC for 36 hours. The following variables were evaluated after 0, 12, 24, and 36 hours refrigeration: total sperm motility, vigor, viability, and plasma and acrosomal membrane integrity. Motility was higher with 2. 5mM of glutathione (57. 8 ± 7. 3) after 12 hours of refrigeration compared to the control (53. 2 ± 8. 3) (P < 0. 05). After 36 hours of refrigeration, motility was higher with 1. 5 mM (43. 4 ± 12. 7) and 2. 5mM glutathione (45. 5 ± 6. 2), than it was with 1mM glutathione (38. 2 ± 9) and the control (35. 5 ± 18. 4) (P < 0. 05), respectively. Vigor was highest with 1. 5mM glutathione (3. 7 ± 0. 3) after 36 hours compared to the control (3. 2 ± 1. 1), (P < 0. 05). Viability differed between control and 1mM treatments (79. 5 ± 1. 8) only after 24 hours (75. 5 ± 9. 7) (P < 0. 05). Throughout the investigation, no significant differences were noted in plasma and acrosomal membrane integrity (P > 0. 05). The 1. 5 and 2. 5mM glutathione levels were more efficient in protecting sperm cells and yielded higher total motility values after 36 hours of refrigeration.

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Effect of storage for 24 h at 18°C on sperm quality and a comparison of two assays for sperm membrane lipid peroxidation

Canadian Journal of Animal Science ◽

10.4141/cjas09122 ◽

2010 ◽

Vol 90 (3) ◽

pp. 389-392 ◽

Cited By ~ 3

Author(s):

N. Am-in ◽

R N Kirkwood ◽

M. Techakumphu ◽

W. Tantasuparuk

Keyword(s):

Lipid Peroxidation ◽

Membrane Permeability ◽

Sperm Motility ◽

Sperm Quality ◽

Membrane Lipid ◽

Membrane Lipid Peroxidation ◽

Group A ◽

Sperm Membrane ◽

Fresh Semen

Boars having normal (71.1 ± 1.2%; n = 10) or low (35.12 &plusmn 3.9%; n = 10) sperm motility 24 h after collection were used, and semen was evaluated following storage in Beltsville Thawing Solution (BTS) for 24 h at 18°C. Sperm lipids were extracted and lipid peroxidation quantified. No differences were evident in fresh semen, but after 24 h, sperm motility, viability and membrane permeability in the low motility group were lower (P < 0.001) compared with the normal motility group. Sperm membrane lipid peroxidation was greater (P < 0.001) in the low motility group. A factor influencing sperm storability is membrane lipid peroxidation, which can be accurately assayed using a commercial kit.Key words: Boars, sperm motility, sperm quality, lipid peroxidation

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Effect of transfection and co-incubation of bovine sperm with exogenous DNA on sperm quality and functional parameters for its use in sperm-mediated gene transfer

Zygote ◽

10.1017/s096719941600037x ◽

2016 ◽

Vol 25 (1) ◽

pp. 85-97 ◽

Cited By ~ 7

Author(s):

María Elena Arias ◽

Esther Sánchez-Villalba ◽

Andrea Delgado ◽

Ricardo Felmer

Keyword(s):

Gene Transfer ◽

Sperm Quality ◽

Computer Assisted ◽

Exogenous Dna ◽

Sperm Analysis ◽

Functional Parameters ◽

Transgenic Embryos ◽

Fertilization Capacity ◽

Motility Parameters

SummarySperm-mediated gene transfer (SMGT) is based on the capacity of sperm to bind exogenous DNA and transfer it into the oocyte during fertilization. In bovines, the progress of this technology has been slow due to the poor reproducibility and efficiency of the production of transgenic embryos. The aim of the present study was to evaluate the effects of different sperm transfection systems on the quality and functional parameters of sperm. Additionally, the ability of sperm to bind and incorporate exogenous DNA was assessed. These analyses were carried out by flow cytometry and confocal fluorescence microscopy, and motility parameters were also evaluated by computer-assisted sperm analysis (CASA). Transfection was carried out using complexes of plasmid DNA with Lipofectamine, SuperFect and TurboFect for 0.5, 1, 2 or 4 h. The results showed that all of the transfection treatments promoted sperm binding and incorporation of exogenous DNA, similar to sperm incorporation of DNA alone, without affecting the viability. Nevertheless, the treatments and incubation times significantly affected the motility parameters, although no effect on the integrity of DNA or the levels of reactive oxygen species (ROS) was observed. Additionally, we observed that transfection using SuperFect and TurboFect negatively affected the acrosome integrity, and TurboFect affected the mitochondrial membrane potential of sperm. In conclusion, we demonstrated binding and incorporation of exogenous DNA by sperm after transfection and confirmed the capacity of sperm to spontaneously incorporate exogenous DNA. These findings will allow the establishment of the most appropriate method [intracytoplasmic sperm injection (ICSI) orin vitrofertilization (IVF)] of generating transgenic embryos via SMGT based on the fertilization capacity of transfected sperm.

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