Decreasing glycerol content by co-supplementation of trehalose and taxifolin hydrate in ram semen extender: Microscopic, oxidative stress, and gene expression analyses

Bucak, Mustafa Numan; Keskin, Nazan; İli, Pınar; Bodu, Mustafa; Akalın, Pınar Peker; Öztürk, Ali Erdem; Özkan, Hüseyin; Topraggaleh, Tohid Rezaei; Sari, Fikret; Başpınar, Nuri; Dursun, Şükrü

doi:10.1016/j.cryobiol.2020.09.001

Cited by 19 publications

(9 citation statements)

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“…The cryoprotective effect of trehalose has been elucidated in studies of sperm from many mammalian species, such as boars ( Gómez-Fernández et al, 2012 ), bulls ( Tuncer et al, 2011 ), rabbits ( Zhu et al, 2017 ), and rams ( Bucak and Tekin, 2007 ; Bucak et al, 2020 ; Bucak et al, 2022 ). Although the exact mechanism of action of trehalose is not fully understood, but it is suggested that trehalose protects cells by increasing the surface tension of the extender and stabilizing the plasma membrane, which is due to the direct interaction of trehalose with the polar headgroups of cell membrane phospholipids ( Iqbal et al, 2018 ), and decrease reactice oxygen species generated and reduce lipid peroxidation during cryopreservation through its own antioxidant effects ( Zhu et al, 2017 ).…”

Section: Discussionmentioning

confidence: 99%

“…Regarding the cryoprotective effect of trehalose on sperm, studies have shown that trehalose cannot enter sperm cells and will form a hyperosmotic environment outside the cell, reduce the formation of intracellular ice crystals, and increase the fluidity of the sperm plasma membrane by reducing the membrane transition temperature. Stabilized biological macromolecules, including proteins, lipids and DNA, effectively protect sperm from cold damage ( Aksan and Toner, 2004 ; Tuncer et al, 2010 ; Iqbal et al, 2017 ; Bucak et al, 2020 ; Olgenblum et al, 2020 ). It has been concluded that the antioxidant properties of trehalose may be related to its effectiveness in membrane cryopreservation ( Lavara et al, 2017 ).…”

Section: Introductionmentioning

confidence: 99%

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Metabolomics analysis of buck semen cryopreserved with trehalose

Wang

et al. 2022

Front. Genet.

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Trehalose is commonly used as an impermeable cryoprotectant for cryopreservation of cells, but its cryoprotective mechanism has now not but been determined. This study investigated the cryopreservation impact of trehalose on buck semen cryopreservation and finished metabolic profiling of freeze-thawed media by way of the GC–MS-based metabolomics for the first time. Metabolic pattern recognition and metabolite identification by means of principal component analysis (PCA), partial least squares discriminant analysis (PLS-DA) and metabolic pathway topology analysis revealed the results of trehalose on buck sperm metabolism at some point of cryopreservation. The results confirmed that trehalose drastically progressed sperm motility parameters and structural integrity after thawing. PCA and PLS-DA analysis discovered that the metabolic patterns of the freezing-thawing media of buck semen cryopreserved with trehalose (T group) or without trehalose (G group, Control) were certainly separated. Using screening conditions of VIP >1.5 and p vaule <0.05, a total of 48 differential metabolites have been recognized, whithin l-isoleucine, L-leucine, L-threonine, and dihydroxyacetone were notably enriched in valine, leucine and isoleucine biosynthesis, glycerolipid metabolism, and aminoacyl-tRNA biosynthesis pathways. In brief, trehalose can efficiently improve membrane structural integrity and motion parameters in buck sperm after thawing, and it exerts a cryoprotective impact with the aid of changing sperm amino acid synthesis and the glycerol metabolism pathway.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Metabolomics analysis of buck semen cryopreserved with trehalose

Wang

et al. 2022

Front. Genet.

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“…19 It has been tested before that reducing the glycerol content from 5% to 3% with 60 mM trehalose supplement increased the post-thawed semen quality. 20 Boron is an element found in soil, rock, nutrients, and water with various applications in industry, agriculture, and medicine. Note that 72.1% of world boron reserves are located in Turkey.…”

Section: Introductionmentioning

confidence: 99%

“…The accessible results of trehalose on post‐thaw semen motility, mitochondrial activity, and acrosome integrity have been also reported in ram semen 19 . It has been tested before that reducing the glycerol content from 5% to 3% with 60 m m trehalose supplement increased the post‐thawed semen quality 20 …”

Section: Introductionmentioning

confidence: 99%

Combination of trehalose and low boron in presence of decreased glycerol improves post‐thawed ram sperm parameters: A model study in boron research

et al. 2021

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Background: Sperm cryopreservation has been widely used in the field of reproductive biotechnology. It applies to certain males of economic and scientific values, including livestock breeds or endangered animal species. The development of a semen extender with a low cryoprotectant concentration and an appropriate amount of trehalose and boron can prevent the deterioration of sperm parameters. Objective:The main goal of this study is to establish a suitable ram extender model, by examining different combinations of high (5%) and low (3%) glycerol concentrations (to reduce its toxic effects on sperm freezing), a fixed amount of trehalose and an increased dose of boron to prevent the deterioration of sperm parameters, and investigate the levels of gene expressions. Materials and methods:The Merino ram ejaculates were collected. The collected ejaculates providing the defined criteria were pooled. The pooled ejaculates were divided into eight aliquots and diluted with the Tris extender including different combinations of glycerol (5% and 3%) and boron (0.25, 0.5, and 1 mM) concentrations and a fixed amount of trehalose, then frozen. After freeze-thawing process, sperm motility, mitochondrial membrane activity, plasma membrane integrity, acrosomal membrane integrity, DNA damage (single cell gel electrophoresis (COMET) and TUNEL assays)

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“…Arsenite can induce the production of ROS by Candida albicans, stimulating the accumulation of trehalose and the activation of catalase in the cells (Urrialde et al, 2017). Trehalose pretreatment effectively prevents the oxidative damage of yeast cells by superoxide (Herdeiro et al, 2006;Uysal et al, 2009;Bucak et al, 2020;Öztürk et al, 2020). Trehalose also protects sperm from oxidative stress and maintains the integrity of the sperm acrosome membrane and plasma membrane (Hu et al, 2010).…”

Section: Introductionmentioning

confidence: 99%

Trehalose can effectively protect sheep epididymis epithelial cells from oxidative stress

et al. 2021

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Abstract. Trehalose, a naturally nontoxic disaccharide that does not exist in mammals, stabilizes cell membrane integrity under oxidative stress conditions, the mechanism of which is still unclear. Here, we analyzed the effects of trehalose on sheep epididymis epithelial cell (EEC) proliferation and its possible mechanisms. To study the effect of trehalose on EECs, EECs were isolated from testes of 12-month-old sheep; cell counting kit-8 (CCK-8) was used to measure the growth of the cells. Cell proliferation was evaluated by assaying cell cycle and apoptosis, and RT-PCR was utilized to identify the epididymal molecular markers glutathione peroxidase 5 (GPX5) and androgen receptor (AR). Next, reactive oxygen species (ROS) content was evaluated by a dichloro-dihydro-fluorescein diacetate (DCFH-DA) probe. Superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) activities were evaluated by enzyme chemistry methods, and GPX5 expression was evaluated by qRT-PCR and enzyme-linked immunosorbent assay (ELISA). The results showed that 100 mM trehalose significantly improved the proliferation potential of EECs, in which the cells could be serially passaged 14 times with continued normal GPX5 and AR marker gene expression in vitro. The trehalose can increase significantly a proportion of EECs in S phase (P<0.01) and decrease significantly the apoptotic rate of EECs (P<0.01) compared to the control. Moreover, the trehalose decreased ROS significantly (P<0.01) and increased CAT (P<0.01) and GSH-Px (P<0.05) activities significantly in EECs. GPX5 mRNA and protein expression were also significantly upregulated in trehalose-treated EECs (P<0.05 and P<0.01 respectively). Our study suggested that exogenous trehalose exhibited antioxidant activity through increasing the activities of CAT, GSH-Px, and the expression level of GPX5 and could be employed to maintain vitality of sheep EECs during long-term in vitro culture.

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Decreasing glycerol content by co-supplementation of trehalose and taxifolin hydrate in ram semen extender: Microscopic, oxidative stress, and gene expression analyses

Cited by 19 publications

References 71 publications

Metabolomics analysis of buck semen cryopreserved with trehalose

Metabolomics analysis of buck semen cryopreserved with trehalose

Combination of trehalose and low boron in presence of decreased glycerol improves post‐thawed ram sperm parameters: A model study in boron research

Trehalose can effectively protect sheep epididymis epithelial cells from oxidative stress

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