Effect of Dimethylacetamide Concentration on Motility, Quality, Antioxidant Biomarkers, Anti-Freeze Gene Expression, and Fertilizing Ability of Frozen/Thawed Rooster Sperm

Mehaisen, Gamal M. K.; Elomda, Ahmed M.; Hamad, Shaimaa K.; Ghaly, Mona M.; Sun, Yanyan; Li, Yunlei; Zong, Yunhe; Chen, Jilan; Partyka, Agnieszka; Nazmi, Ali; Abbas, Ahmed O.; Stino, F. K. R.

doi:10.3390/ani12202739

Cited by 12 publications

(5 citation statements)

References 55 publications

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“…Qi et al [27] reported that RPL29 expression in rooster sperm was substantially downregulated following cryopreservation. However, our study did not show any significant effect on the RPL29 expression using various levels of DMA in postthawed sperm, and these results are in agreement with other studies [2]. On the other hand, Riesco and Robles [56] reported that cryopreservation of Zebrafish genital ridges downregulated most of the gene transcripts and demonstrated that this effect was a result of the freezing/thawing procedure rather than exposure to CPAs themselves.…”

Section: Discussionsupporting

confidence: 92%

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The In Vitro Evaluation of Rooster Semen Pellets Frozen with Dimethylacetamide

Hamad

Elomda

Sun

et al. 2023

Animals

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Sperm cryopreservation is an effective technique for conserving animal genetic diversity and transmitting superior genetic backgrounds, maintained via a non-invasive sampling and collection of huge quantities of sperm. Nevertheless, cryopreservation in avian species is not commercially viable because of the rooster sperm’s susceptibility to damage. This study aims to estimate the impact of dimethylacetamide (DMA) as a cryoprotectant at different levels (3%, 6%, or 9%) on the post-thawed sperm quality, motility, antioxidant-biomarkers, and the expression of anti-freeze related genes. Semen samples were collected twice a week from twelve roosters aged 40 wk, weighing 3400 ± 70 g, and belonging to the Cairo-B2 chicken strain. Fresh semen samples were rapidly appraised, pooled, diluted with two volumes of a basic extender, and divided equally into three groups. The diluted groups were chilled at −20 °C for 7 min, then gently supplemented with 3, 6, or 9% pre-cooled DMA and equilibrated at 5 °C for a further 10 min. Semen pellets were formed by pipetting drops 7 cm above liquid nitrogen (LN2), which were then kept inside cryovials in the LN2. Thawing was performed 2 months later by taking 3–4 pellets of the frozen semen into a glass tube and warming it in a water bath for 8 s at 60 °C. The results showed that 3% DMA increased the proportion of total motile sperm, progressivity, viability, and plasma membrane integrity (%) compared to the 6% and 9% DMA groups. The lipid peroxidation and antioxidant enzyme activity were improved in the 3% group. At the same time, some anti-freeze-related genes’ (including ras homolog family member A (RHOA), heat shock protein 70 (HSP70), and small nuclear ribonucleoprotein polypeptide A (SNRPA1)) expressions were upregulated within the 3% DMA group relative to other groups. In conclusion, the 3% DMA group maintained higher post-thawed sperm quality than the other tested groups.

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Section: Discussionsupporting

confidence: 92%

“…Rooster spermatozoa have smaller cytoplasm contents, antioxidants, and mitochondria and higher polyunsaturated fatty acid levels within the plasma membrane than their counterpart in mammals. Thus, rooster spermatozoa are more susceptible to damage throughout the freezing-thawing process [1,2].…”

Section: Introductionmentioning

confidence: 99%

The In Vitro Evaluation of Rooster Semen Pellets Frozen with Dimethylacetamide

Hamad

Elomda

Sun

et al. 2023

Animals

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“…Despite difficulties, successful cryopreservation protocols have been developed for valuable genetic lines of chickens, turkeys, ducks and geese using glycerol or dimethylacetamide (DMA) as permeating cryoprotectants 6 – 8 . Optimal cooling rates, straw sizes and thawing rates have been elucidated 4 , 8 , 9 . Non-permeating agents like liposomes, egg yolk or polymers provide additional sperm protection when added to standard glycerol or DMA freezing media 10 – 12 .…”

Section: Introductionmentioning

confidence: 99%

Apigenin supplementation substantially improves rooster sperm freezability and post-thaw function

Najafi,

Mohammadi,

Sharifi

et al. 2024

Sci Rep

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This pioneering research investigated apigenin potential to augment rooster sperm cryosurvival in an extender model. Apigenin is a natural antioxidant flavonoid showing promise for improved post-thaw sperm function. However, its effects on avian semen cryopreservation remain unexplored. This first study supplemented rooster sperm Lake extender with 0, 50, 100, 200, 400 μmol/L apigenin to determine the optimal concentrations for post-thaw quality. Supplementation with 100 μmol/L apigenin resulted in significant enhancements in total motility (from 41.5% up to 71.5%), progressive motility (18.1% to 29.1%) (p < 0.05), membrane integrity (40% to 68%), mitochondrial function (p < 0.001), viability (37% to 62%) and total antioxidant capacity (p < 0.001) compared to the control. It also substantially reduced percentages of abnormal morphology, reactive oxygen species and apoptosis (p < 0.001). Although 200 μmol/L apigenin significantly enhanced some attributes, effects were markedly lower than 100 μmol/L. Higher doses did not improve cryoprotective parameters. This indicates 100 μmol/L as the optimal apigenin concentration. This represents the first report of apigenin protecting rooster sperm from cryodamage. The natural antioxidant improved post-thaw sperm quality, likely by suppressing oxidative stress and apoptosis. Apigenin shows promise for enhancing rooster sperm cryosurvival.

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“…The previous study analysed the antioxidant biomarkers and the anti-freeze-associated gene expression in the postthawed sperm of rooster (Mehaisen et al 2022). Testes are the site of spermatozoa production, while the epididymis has stored spermatozoa (Hedger 2011).…”

Section: Introductionmentioning

confidence: 99%

Differential expression and localisation of heat shock protein 70 (HSP70) and glutathione peroxidase 5 (GPX5) in the testis and epididymis of Sonid Bactrian camels

Hasi

Sodnompil

et al. 2023

Reprod. Fertil. Dev.

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Context Heat shock protein 70 (HSP70) and glutathione peroxidase 5 (GPX5) are biomarkers of oxidative stress and stress in temperate, tropical environments, which are crucial for male reproduction. Their expression and distribution patterns in the testis and epididymis of Bactrian camels are still unknown. Aims This study aims to investigate the HSP70 and GPX5 expression and localisation in 3- and 6-year-old Bactrian camel testis and epididymis. Methods Reverse transcription quantitative polymerase chain reaction (qRT-PCR), Western blot and immunohistochemistry were used to detect HSP70 in the testis and epididymis (caput, corpus and cauda) and GPX5 in the epididymis at two developmental stages (3-year-old puberty group and 6-year-old adult group). Key results HSP70 was upregulated in the testis. Immunohistochemistry results indicated the HSP70 protein was mainly detected in spermatids and Leydig cells of testicular tissue. In the epididymis, HSP70 was located at the luminal spermatozoa, the epithelium lining the epididymal and the epididymal interstitium. GPX5 expression was significantly higher in the caput epididymis than in the corpus and cauda epididymis. GPX5 protein was observed in the epithelium lining the epididymal, interstitium and luminal spermatozoa in the epididymis by immunohistochemistry. Conclusions Bactrian camel HSP70 and GPX5 exhibited spatiotemporal expression specificity. Implications HSP70 and GPX5 may be essential for germ cell development and reproductive success after sexual maturation in Sonid Bactrian camels.

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Effect of Dimethylacetamide Concentration on Motility, Quality, Antioxidant Biomarkers, Anti-Freeze Gene Expression, and Fertilizing Ability of Frozen/Thawed Rooster Sperm

Cited by 12 publications

References 55 publications

The In Vitro Evaluation of Rooster Semen Pellets Frozen with Dimethylacetamide

The In Vitro Evaluation of Rooster Semen Pellets Frozen with Dimethylacetamide

Apigenin supplementation substantially improves rooster sperm freezability and post-thaw function

Differential expression and localisation of heat shock protein 70 (HSP70) and glutathione peroxidase 5 (GPX5) in the testis and epididymis of Sonid Bactrian camels

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