Melatonin can improve viability and functional integrity of cooled and frozen/thawed rabbit spermatozoa

Fadl, Aya M.; Ghallab, A. M.; Abou-Ahmed, M.M.; Moawad, Adel R.

doi:10.1111/rda.13853

Cited by 16 publications

(21 citation statements)

References 63 publications

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“…In addition, the highest degree of progressive motility corresponded to this same concentration of MT; however, no significant difference was observed in the frozen control group. The results of this part of the study are consistent with those of a study by Fadl et al [ 18 ] that found that concentrations of 1 mM MT corresponded to the highest degree of progressive motility compared to other doses as well as the control group. In addition, the present study showed that adding Se to the extender did not significantly improve rat sperm motility after thawing.…”

Section: Discussionsupporting

confidence: 92%

“…Semen samples with a concentration of 20×10 6 sperm/mL to 30×10 6 sperm/mL were diluted in a TL-HEPES extender solution, and the semen of each rat was divided into six groups, with four groups containing MT and Se at concentrations of 0.5 mM and 1 mM and two groups containing no added material, which were used as fresh and cryopreservation control groups, respectively [ 18 , 19 ]. The samples were cooled to 4°C and equilibrated at this temperature for 2 hours.…”

Section: Methodsmentioning

confidence: 99%

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Melatonin and selenium supplementation in extenders improves the post-thaw quality parameters of rat sperm

Shahandeh

Ghorbani

Mokhlesabadifarahani

et al. 2022

Clin Exp Reprod Med

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Objective: The aim of this study was to determine the effects of melatonin and selenium in freezing extenders on frozen-thawed rat sperm.Methods: Semen samples were collected from 20 adult male Wistar albino rats. Following dilution, the samples were divided into six groups: four cryopreserved groups with 1 mM and 0.5 mM melatonin and selenium supplements, and two fresh and cryopreserved control groups. The rapid freezing technique was used to freeze the samples. Flow cytometry was used to assess plasma membrane integrity, mitochondrial membrane potential, and DNA damage, while computer-assisted sperm analysis was used to assess motility.Results: Total motility was higher in the 1 mM melatonin supplementation group than in the cryopreserved control group (mean±standard error of the mean, 69.89±3.05 vs. 59.21±1.31; p≤0.05). The group with 1 mM selenium had the highest plasma membrane integrity (42.35%±1.01%). The cryopreserved group with 0.5 mM selenium had the highest mitochondrial membrane potential, whereas the cryopreserved control group had the lowest (45.92%±4.53% and 39.45%±3.52%, respectively).Conclusion: Cryopreservation of rat semen supplemented with 1 mM melatonin increased sperm motility after freeze-thawing, while supplementation with 0.5 mM selenium increased mitochondrial activity.

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Section: Discussionsupporting

confidence: 92%

Section: Methodsmentioning

confidence: 99%

Melatonin and selenium supplementation in extenders improves the post-thaw quality parameters of rat sperm

Shahandeh

Ghorbani

Mokhlesabadifarahani

et al. 2022

Clin Exp Reprod Med

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“…Melatonin, as an antioxidant, anti-apoptotic and free-radical scavenger, is involved in cell protection [ 20 ]. In recent years, the protective effect of adding melatonin to the freezing medium has been tested on sperm viability after thawing [ 38 – 40 ]. The results demonstrated that melatonin at a concentration of 100 μM increased the viability of frozen–thawed SSCs.…”

Section: Discussionmentioning

confidence: 99%

Melatonin in cryopreservation media improves transplantation efficiency of frozen–thawed spermatogonial stem cells into testes of azoospermic mice

et al. 2022

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Background Cryostorage of spermatogonial stem cells (SSCs) is an appropriate procedure for long-term storage of SSCs for fertility preservation. However, it causes damage to cellular structures through overproduction of ROS and oxidative stress. In this study, we examined the protective effect of melatonin as a potent antioxidant in the basic freezing medium to establish an optimal cryopreservation method for SSCs. Methods SSCs were obtained from the testes of neonatal male mice aged 3–6 days. Then, 100 μM melatonin was added to the basic freezing medium containing DMSO for cryopreservation of SSCs. Viability, apoptosis-related markers (BAX and BCL2), and intracellular ROS generation level were measured in frozen–thawed SSCs before transplantation using the MTT assay, immunocytochemistry, and flow cytometry, respectively. In addition, Western blotting and immunofluorescence were used to evaluate the expression of proliferation (PLZF and GFRα1) and differentiation (Stra8 and SCP3) proteins in frozen–thawed SSCs after transplantation into recipient testes. Results The data showed that adding melatonin to the cryopreservation medium markedly increased the viability and reduced intracellular ROS generation and apoptosis (by decreasing BAX and increasing BCL2) in the frozen–thawed SSCs (p < 0.05). The expression levels of proliferation (PLZF and GFRα1) and differentiation (Stra8 and SCP3) proteins and resumption of spermatogenesis from frozen–thawed SSCs followed the same pattern after transplantation. Conclusions The results of this study revealed that adding melatonin as an antioxidant to the cryopreservation medium containing DMSO could be a promising strategy for cryopreservation of SSCs to maintain fertility in prepubertal male children who suffer from cancer.

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“…Zhu and colleague investigated the effects of supplementation of amin E analogue [ 64 ], cysteine [ 65 ], glutamine [ 66 ], trehalose [ 67 ] and melatonin [ 68 ], and revealed that supplementation of these antioxidative agents in Tris-citrate-glucose extender decreased ROS levels and improved the quality of frozen–thawed rabbit sperms. Fadl et al confirmed 1.0 mM melatonin improved the motility, viability, membrane and acrosome integrities, and DNA integrity of frozen–thawed rabbit sperms in different extender (INRA-82) [ 69 ] Additionally, curcumin and curcumin nanoparticles were confirmed to improve the post-thawed quality of rabbit sperms via redox signaling and reduce the apoptosis process [ 70 ]. On the other hand, Maya-Soriano et al reported that supplementation of bovine serum albumin, retinol and retinyl in the sperm freezing extender has no beneficial effect on the viability, mortality, progressivity, and acrosome and morphological integrity of frozen–thawed rabbit sperms at the concentrations they tested [ 71 ].…”

Section: Effects Of Supplementsmentioning

confidence: 99%

Strategies for Highly Efficient Rabbit Sperm Cryopreservation

Nishijima

Kitajima

Matsuhisa

et al. 2021

Animals

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The rabbit is a valuable animal for both the economy and biomedical sciences. Sperm cryopreservation is one of the most efficient ways to preserve rabbit strains because it is easy to collect ejaculate repeatedly from a single male and inseminate artificially into multiple females. During the cooling, freezing and thawing process of sperms, the plasma membrane, cytoplasm and genome structures could be damaged by osmotic stress, cold shock, intracellular ice crystal formation, and excessive production of reactive oxygen species. In this review, we will discuss the progress made during the past years regarding efforts to minimize the cell damage in rabbit sperms, including freezing extender, cryoprotectants, supplements, and procedures.

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Melatonin can improve viability and functional integrity of cooled and frozen/thawed rabbit spermatozoa

Cited by 16 publications

References 63 publications

Melatonin and selenium supplementation in extenders improves the post-thaw quality parameters of rat sperm

Melatonin and selenium supplementation in extenders improves the post-thaw quality parameters of rat sperm

Melatonin in cryopreservation media improves transplantation efficiency of frozen–thawed spermatogonial stem cells into testes of azoospermic mice

Strategies for Highly Efficient Rabbit Sperm Cryopreservation

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