Effects of kinetin supplementation on the post-thaw motility, viability, and structural integrity of dog sperm

Qamar, Ahmad Yar; Fang, Xun; Bang, Seonggyu; Kim, Min Jung; Cho, Jongki

doi:10.1016/j.cryobiol.2020.05.015

Cited by 16 publications

(17 citation statements)

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“…Many investigators have focused on the use of antioxidants in the freezing media to reduce the negative effects of ROS on sperm [ 2 , 4 , 10 ]. Our previous findings showed that supplementation of dog cryopreservation medium with kinetin [ 42 ] and astaxanthin [ 41 ] improved the post-thaw motility, viability, and structural integrity of dog sperm, and we aimed to assess the effect of resveratrol (RSV) as an alternative antioxidant.…”

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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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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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“…Antioxidantsupplemented sperm showed reduced lipid peroxidation, enabling the plasma membrane to maintain normal physiological and metabolic activity, ultimately resulting in enhanced viability (Alvarez &Storey 1989). Several studies reported the useful effects of antioxidants during semen freezing to minimize the adverse impacts of ROS on spermatozoa has been reported, thereby improving post-thaw quality (Bansal &Bilaspuri 2010, Malo et al 2010, Qamar et al 2020, Setyawan et al 2016, Yoshimoto et al 2008). However, nding an appropriate species-speci c antioxidant is the target of several research groups to alleviate the cryodamage and maintaining functional integrity of spermatozoa during freezing process (Bansal &Bilaspuri 2010).…”

Section: Introductionmentioning

confidence: 99%

“…However, nding an appropriate species-speci c antioxidant is the target of several research groups to alleviate the cryodamage and maintaining functional integrity of spermatozoa during freezing process (Bansal &Bilaspuri 2010). For instance, α-Linoleic acid was shown to suppress ROS generation by stabilizing the plasma membrane during the cryopreservation of boar sperm (Qamar et al 2020). In canine species, several antioxidants and ROS scavenger supplements were used for sperm cryopreservation, including rosemary and spermine (Setyawan et al 2016, Vieira et al 2018).…”

Section: Introductionmentioning

confidence: 99%

“…Arti cial insemination using frozen dog semen is associated with a sluggish progress compared with that of other animal species (England 1993). Identifying the best cryoprotectant for preventing cryodamage-induced molecular toxicity in dog sperm post-thaw is a focus for several research groups, including our group (Farstad 2009, Qamar et al 2020). Reactive oxygen species (ROS) are resulted from physiological cellular metabolic processes, which pushes the cells to early aging and apoptosis (Redza-Dutordoir &Averill-Bates 2016).…”

Section: Introductionmentioning

confidence: 99%

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Quercetin Improves the Apoptotic Index and Oxidative Stress in Post-Thaw Dog Sperm

Bang

Qamar

Tanga

et al. 2021

Preprint

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This study was conducted to investigate if quercetin (QRN) may ameliorate apoptosis and oxidative stress in post-thaw dog sperm. Herein, we evaluated the post-thaw apoptosis and oxidative stress after treatment with QRN (control, 25, 50, and 100 µM) in freezing of dog semen. The oxidative stress index was significantly affected (p<0.05) between the various concentrations of QRN and the control (17.56 ± 1.02, 7.54 ± 0.48, 5.66 ±0.80, and 10.41 ± 0.69), respectively. The apoptosis index was 9.1 ± 1.34, 6.66 ± 0.58, 6.77 ± 0.66, and 5.38 ± 0.86 in the control, and 25, 50, and 100 µM QRN treatment groups, respectively (p< 0.05). The effects of ameliorated cryo-induced damage by QRN on post-thaw sperm quality were also observed through improved structural and functional tests. Sperm treated with 50 µM QRN showed significantly higher motility (51.8 ± 2.1% vs. 43.1 ± 1.4%, P < 0.05), survival rates (46.9 ± 0.7% vs. 43.9 ± 0.4%, P < 0.05), and mucus penetration than control group, respectively. Results demonstrate that supplementing freezing buffer with 50 µM QRN reduced oxidative damage and improved the quality of post-thaw dog sperm.

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“…However, some antioxidants have pro-oxidant effects, depending on their concentration and neighboring molecules [9,10]. Thus, unconventional antioxidants such as curcumin, kinetin, and quercetin have attracted interest in terms of improving the quality of thawed canine sperm, increasing its antioxidant capacity, and of expressing genes that modulate reactive oxygen species in sperm [11][12][13]. Therefore, we investigated the effects of 2-mercaptohistidine trimethylbetaine (ergothioneine (ERG)) and 2-isopropyl-3,6-dimethoxy-5-methylphenol (isoespintanol (ISO)), which have high antioxidant capacity and mitigate alterations in cryopreserved semen from other animal species [14][15][16].…”

Section: Introductionmentioning

confidence: 99%

Cryoprotective Effects of Ergothioneine and Isoespintanol on Canine Semen

Suarez

Tejera

Gómez

et al. 2021

Animals

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Sperm undergo oxidative stress due to excessive production of reactive oxygen species (ROS) during cryopreservation. Some unconventional natural antioxidants can reduce ROS-induced changes in cryopreserved canine sperm. This study aimed to identify the cryoprotective effects of ergothioneine and isoespintanol on the quality of thawed canine semen. Twelve ejaculates from six dogs were cryopreserved in a tris-yolk extender without (control) or with 50 (E50), 100 (E100), or 150 (E150) µM ergothioneine or 20 (I20), 40 (I40), or 60 (I60) µM isoespintanol. We evaluated the motility and kinetics of thawed sperm using computerized analysis; determined morphology by eosin-nigrosin staining; functional membrane integrity using hypoosmotic tests, and structural membrane and acrosome integrity; mitochondrial membrane potential by fluorescence microscopy; and ROS production by spectrophotometry. Data were statistically analyzed using mixed models and Tukey tests. E100 increased total (60.6% vs. 49.6%) and progressive (26.4% vs. 20.1%) motility, straight line velocity (41.3 vs. 35.9 µm/s), and rapid sperm (17.6% vs. 12.3%) compared with controls. However, E150 reduced the numbers of hyperactive sperm. E100, I40, and I60 reduced the abnormal morphology and ROS production, and all concentrations of both antioxidants increased acrosomal integrity. We concluded that ergothioneine and isoespintanol reduce deleterious sperm alterations and oxidative stress in thawed canine semen.

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Effects of kinetin supplementation on the post-thaw motility, viability, and structural integrity of dog sperm

Cited by 16 publications

References 61 publications

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

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

Quercetin Improves the Apoptotic Index and Oxidative Stress in Post-Thaw Dog Sperm

Cryoprotective Effects of Ergothioneine and Isoespintanol on Canine Semen

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