Effects of Discontinuous Percoll Gradient Containing Alpha-linolenic Acid on Characteristics of Frozen-thawed Boar Spermatozoa

Kim, Doo-San; Hwangbo, Yong; Cheong, Hyeonsook; Park, Choon‐Keun

doi:10.12750/jarb.35.1.58

Cited by 7 publications

(5 citation statements)

References 22 publications

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“…The plasma membrane integrity of sperm was evaluated using sperm SYBR14 and PI doubling staining method (Kim et al, 2020). The sperm samples of 7.5 × 10 6 sperm/mL were diluted with 40 nM SYBR-14 and 2.0 μM PI which were incubated at 38℃ for 5 min at dark room and then, and these samples were centrifuged to remove supernatants at 410 g for 5 min.…”

Section: Detection Of Plasma Membrane Integritymentioning

confidence: 99%

Effect of bicarbonate and progesterone on plasma membrane integrity, acrosome reaction and proportion of fatty acids in boar sperm

Park

Lee

2022

J Anim Reprod Biotechnol

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This study investigated the influence of sodium bicarbonate (NaHCO 3 ) and progesterone on acrosome reaction and proportion of polyunsaturated fatty acid (PUFA) composition boar sperm. The sperm were diluted with semen extender and incubated with NaHCO 3 and progesterone at 38℃, 5% CO 2 for 6 h. Plasma membrane integrity and acrosome reaction were analyzed using SYBR14/propidium iodide (PI) and FITC-PNA/PI doubling staining method, and proportion of PUFA was analyzed using gas chromatography. In results, Plasma membrane integrity was significantly decreased in 50 mM NaHCO 3 group and acrosome reaction was significantly increased by over the 100 mM NaHCO 3 group compared to control group (p < 0.05). In addition, progesterone significantly increased decreased plasma membrane integrity at 100 mM progesterone and acrosome reaction at over the 5.0 μM progesterone (p < 0.05), but there was no difference among the 5.0 to 100 μM groups. PUFAs were significantly decreased in 100 mM NaHCO 3 and 50 μM progesterone treatments compared to control group. In summary NaHCO 3 and progesterone induce acrosome reaction and reduce PUFA composition in boar sperm, therefore, the results maybe help to understand basically knowledge for the acrosome reaction and PUFA composition in boar sperm.

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Section: Detection Of Plasma Membrane Integritymentioning

confidence: 99%

Effect of bicarbonate and progesterone on plasma membrane integrity, acrosome reaction and proportion of fatty acids in boar sperm

Park

Lee

2022

J Anim Reprod Biotechnol

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“…The novel idea of the possibility to preserve the genetic materials enables the conservation of endangered animal species, enables artificial reproduction, and the decision of breeding the required pedigree [8,20,23]. The cryopreservation method of genetic conservation, commonly employed in livestock reproduction and conservation, has various limitations in terms of maintaining the fertility parameters and one of the common limitations is cryo-induced damage [7,9,33,49].…”

Section: Introductionmentioning

confidence: 99%

Resveratrol supplementation into extender protects against cryodamage in dog post-thaw sperm

Bang

Qamar

Tanga

et al. 2021

The Journal of Veterinary Medical Science

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Antioxidants have multiple protective roles in a variety of cells and thus can be used to protect sperm against cryo-damage during freezing, which affects fertility. The antioxidant resveratrol (3,5,4-trihydroxytrans-stilbene; RSV) has been reported to protect the animal sperm during cryopreservation, including human sperm. In this study, we assessed the protective effects of RSV supplementation on dog sperm cryopreservation. Semen was collected from four dogs and the effect of different concentrations of RSV (0, 100, 200, and 400 µM) on post-thaw sperm quality was examined. After thawing, sperm motility was assessed using computer-aided sperm analysis, and the structural integrity of the plasma membrane, acrosome, and chromatin was examined. In addition, their mitochondrial activity and gene expression were also assessed. Dog sperm cryopreserved with 200 µM RSV showed significant improvement in post-thaw sperm motility and viability compared with that of the control group ( P <0.05). Moreover, RSV-supplemented samples showed significantly higher numbers of sperm with an intact plasma membrane, active mitochondria, and structural integrity of acrosomes and chromatin than that of control samples ( P <0.05). Furthermore, gene expression showed that RSV supplemented samples showed lower expression of pro-apoptotic ( BAX ), reactive oxygen species (ROS) modulator oxidative stress-related ( ROMO1 ) and 8-oxoguanine DNA glycosylase 1 ( OGG1 ) whereas higher expression levels of anti‐apoptotic ( BCL2 ), protamine-2 ( PRM2 ), protamine-3 ( PRM3 ) and sperm acrosome‐associated 3 ( SPACA3 ) genes than control. Our results suggest that RSV, at its optimum concentration, can be efficiently used as an antioxidant in the cryopreservation of dog sperm.

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“…Sperm freezing was performed according to Kim et al [ 37 ]. In brief, pooled semen samples were washed using centrifugation at 500 g with basic diluent supplemented with D(+)-Glucose (6.00 g/L), EDTA (0.45g/L), sodium citrate (1.38g/L), sodium bicarbonate (0.2g/L), Trizma base (1.00 g/L), citric acid (0.5g/L), cysteine (0.01g/L), BSA (0.8g/L), and kanamycin sulfate (0.06g/L).…”

Section: Methodsmentioning

confidence: 99%

Cryopreservation of Pig Semen Using a Quercetin-Supplemented Freezing Extender

Bang

Tanga

Fang

et al. 2022

Life

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Reactive oxygen species (ROS) produced during freeze–thaw procedures cause oxidative damage to the sperm, reducing fertility. We aimed to improve the post-thaw quality of pig sperm by quercetin (QRN) supplementation to reduce the cryodamage associated with the freeze–thaw procedure. Four equal aliquots of pooled boar semen were diluted with a freezing extender supplemented with different concentrations of QRN (0, 25, 50, and 100 µM) and then were subjected to cryopreservation in liquid nitrogen. Semen analysis was performed following 7 days of cryopreservation. Results demonstrated that the semen samples supplemented with 50 µM QRN significantly improved the post-thaw sperm quality than those subjected to other supplementations (p < 0.05). Semen samples supplemented with 50 µM QRN showed significantly improved plasma membrane functional integrity (47.5 ± 1.4 vs. 43.1 ± 4.1, 45.3 ± 1.7, and 44.1 ± 1.4) and acrosome integrity (73.6 ± 3.4 vs. 66.3 ± 2.4, 66.7 ± 3.6, and 68.3 ± 32.9) as compared to the control, 25 µM, and 100 µM QRN groups, respectively. The mitochondrial activity of the 50 µM QRN group was greater than control and 25 µM QRN groups (43.0 ± 1.0 vs. 39.1 ± 0.9 and 41.9 ± 1.0) but showed no difference with the 100 µM QRN group. Moreover, the 50 µM QRN group showed a higher sperm number displaced to 1 cm and 3 cm points in the artificial mucus than other groups. Therefore, supplementing the freezing extender with QRN can serve as an effective tool to reduce the magnitude of oxidative damage associated with sperm freezing.

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Effects of Discontinuous Percoll Gradient Containing Alpha-linolenic Acid on Characteristics of Frozen-thawed Boar Spermatozoa

Cited by 7 publications

References 22 publications

Effect of bicarbonate and progesterone on plasma membrane integrity, acrosome reaction and proportion of fatty acids in boar sperm

Effect of bicarbonate and progesterone on plasma membrane integrity, acrosome reaction and proportion of fatty acids in boar sperm

Resveratrol supplementation into extender protects against cryodamage in dog post-thaw sperm

Cryopreservation of Pig Semen Using a Quercetin-Supplemented Freezing Extender

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