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This study aimed to investigate the morphological, functional and molecular changes in frozen–thawed ram sperm using an extender containing different concentrations of hydrated carbon 60 fullerene (C60HyFn), a nanotechnological product. Semen taken from each of the seven Akkaraman rams were pooled. Semen collection was done twice a week and it continued for 3 weeks. Each pooled semen sample was divided into six equal groups and diluted with tris + egg yolk extender including 0 (control), 200, 400, 800 nM, 1 and 5 μM concentrations of C60HyFn at 37°C. They were then frozen in liquid nitrogen vapour at −140°C, stored in liquid nitrogen container (−196°C) and thawed at 37°C for 25 s before analysis. In comparison with control, C60HyFn addition prior to freezing procedure provided significant increases in total and progressive motility rates, glutathione peroxidase, catalase activities and percentage of highly active mitochondria, and significant decreases in dead and abnormal sperm rates, lipid peroxidation, caspase‐3 and DNA fragmentation levels in frozen–thawed ram semen. When compared to control, C60HyFn supplementation significantly down‐regulated the expression levels of miR‐200a and KCNJ11, and significantly up‐regulated the expression levels of miR‐3958‐3p (at the concentrations of 200, 400, 800 nM and 1 μM), CatSper1 (at the concentrations of 200, 400 nM and 5 μM), CatSper2 (at the concentrations of 1 and 5 μM), CatSper3 (at the concentrations of 200, 400 nM, 1 and 5 μM), CatSper4 (at all concentrations), ANO1 (at the concentrations of 800 nM, 1 and 5 μM) and TRPV5 (at the concentrations of 200, 400 and 800 nM). The addition of C60HyFn had no effect on global DNA methylation rates. As a result, C60HyFn supplementation to ram semen extenders may be beneficial in reducing some of the functional, structural and molecular damages in sperm induced by the freeze–thawing procedure. [ABSTRACT FROM AUTHOR]

This study was carried out to investigate the effect of the semen freeze-thawing process on the functionality and molecular structure of ram spermatozoa. The temperature of pooled and diluted semen at 38°C (group 1, control) was lowered to 5°C (group 2), and it was subjected to glycerolisation-equilibration (group 3), frozen and thawed (group 4). Compared to the control, deterioration in spermatological parameters and significant increases in lipid peroxidation and global DNA methylation levels were observed in groups 3 and 4. When compared with the control, significant downregulation in the levels of miR-485 of group 2, miR-29a of group 3 and let-7a, miR-485 and miR-29a of group 4, and significant upregulation in the levels of miR-107 of group 3 and miR-127 of groups 3 and 4 were detected. In comparison to the control, significant upregulation in the levels of CatSper1, CatSper2, CatSper3, CatSper4, ANO1 and TRPM3 of group 2, CatSper4, ANO1 and TRPM3 of group 3 and KCNJ11 of group 4, and significant downregulation in the CatSper 3 level of group 4 were determined. As a result, the semen freeze-thawing process causes motility and morphological disorders in rams. This may be due to molecular changes associated with lipid peroxidation in spermatozoa. [ABSTRACT FROM AUTHOR]

The aim of this study was to determine the effect of different doses of hydrated C60 fullerene (C60HyFn) nanoparticles on ram semen during cool storage at 5 °C. Semen samples were collected from 7 healthy Akkaraman rams by artificial vagina during nonbreeding season (April) 3 times for two weeks (3 replications), and samples having good initial sperm motility (> 70 %) and count (> 2 billion / mL) were pooled and diluted with tris-egg yolk based extender at 37 °C. Then, the diluted semen samples were divided into 15 aliquots control and 14 different C60HyFn groups (25 nM, 50 nM, 100 nM, 200 nM, 400 nM, 800 nM, 1 µM, 5 µM, 10 µM, 20 µM, 40 µM, 60 µM, 80 µM, and 100 µM), and stored at 5 °C up to 144th h. The motility (total, progressive, rapid, medium, slow, and static), kinematic (VCL, VSL, VAP, LIN, STR, WOB, ALH, and BCF), and some other spermatological parameters (dead, abnormal, damaged acrosome, and membrane intactness) were examined at 0, 12, 24, 48, 72, 96, 120, and 144th h. All C60HyFn doses except 25 nM, 50 nM, 60 µM, 80 µM, and 100 µM had numerically higher total and progressive motility, membrane intactness and numerically lower static, dead and total abnormal spermatozoon ratios at the end of 144th h than control. The differences between 400 nM dose and control in membrane intactness, and also between 400, 800 nM doses and control in dead spermatozoon ratio were statistically significant (P < 0.0125). In conclusion, it is suggested that supplementation of any doses of C60HyFn between 100 nM and 40 µM may be beneficial to keep some spermatological parameters at high level during the short-term storage of ram semen at 5 °C. [ABSTRACT FROM AUTHOR]

The aim of this study was to investigate the effect of Michauxia campanuloides L'Hér. on some semen parameters in rats. The plant material of the study is Michauxia campanuloides L'Hér. having antioxidant property. The animal material of the study consisted of 18 male Wistar albino rats. The rats were divided into three groups as control (C; n=6), treatment 1 (T1; n=6) and treatment 2 (T2; n=6). Lyophilized aqueous extract of M. campanuloides L'Hér. was administered orally at a dose of 20 mg/kg/day for rats in the T1 group and 40 mg/kg/day for rats in the T2 group for 21 days. At the end of the study, all rats were examined for blood total antioxidant capacity (TAC), semen density, semen motility, abnormal spermatozoon ratio along with testicular, epididymal, seminal gland and ventral prostate weight. Semen density was significantly higher in the T2 group compared to the C and T1 groups (P=0.002). Rates of abnormal spermatozoa were significantly lower in T1 and T2 groups than the C group (P=0.003). The weight of seminal gland was significantly higher in the T1 and T2 groups compared to the C group (P=0.016). In conclusion, oral consumption of M. campanuloides extract provides an increase in semen quality in rats. [ABSTRACT FROM AUTHOR]

This study aimed to investigate the morphological, functional and molecular changes in frozen-thawed ram sperm using an extender containing different concentrations of hydrated carbon 60 fullerene (C 60 HyFn), a nanotechnological product. Semen taken from each of the seven Akkaraman rams were pooled. Semen collection was done twice a week and it continued for 3 weeks. Each pooled semen sample was divided into six equal groups and diluted with tris + egg yolk extender including 0 (control), 200, 400, 800 nM, 1 and 5 μM concentrations of C 60 HyFn at 37°C. They were then frozen in liquid nitrogen vapour at -140°C, stored in liquid nitrogen container (-196°C) and thawed at 37°C for 25 s before analysis. In comparison with control, C 60 HyFn addition prior to freezing procedure provided significant increases in total and progressive motility rates, glutathione peroxidase, catalase activities and percentage of highly active mitochondria, and significant decreases in dead and abnormal sperm rates, lipid peroxidation, caspase-3 and DNA fragmentation levels in frozen-thawed ram semen. When compared to control, C 60 HyFn supplementation significantly down-regulated the expression levels of miR-200a and KCNJ11, and significantly up-regulated the expression levels of miR-3958-3p (at the concentrations of 200, 400, 800 nM and 1 μM), CatSper1 (at the concentrations of 200, 400 nM and 5 μM), CatSper2 (at the concentrations of 1 and 5 μM), CatSper3 (at the concentrations of 200, 400 nM, 1 and 5 μM), CatSper4 (at all concentrations), ANO1 (at the concentrations of 800 nM, 1 and 5 μM) and TRPV5 (at the concentrations of 200, 400 and 800 nM). The addition of C 60 HyFn had no effect on global DNA methylation rates. As a result, C 60 HyFn supplementation to ram semen extenders may be beneficial in reducing some of the functional, structural and molecular damages in sperm induced by the freeze-thawing procedure., (© 2023 Wiley-VCH GmbH. Published by John Wiley & Sons Ltd.)