Cholesterol addition protects membrane intactness during cryopreservation of stallion sperm

Oliveira, C.H.; Vasconcelos, André Belico de; Souza, Fernando Andrade; Martins‐Filho, Olindo Assis; Silva, M.X.; Varago, F. C.; Lagares, M. A.

doi:10.1016/j.anireprosci.2009.08.011

Cited by 41 publications

(29 citation statements)

References 29 publications

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“…However, the C/P ratio in cattle bull spermatozoa varied from 0.42 to 0.45 [20]. There is an active participation of sperm plasma membrane in the process of capacitation, mainly through loss of cholesterol [5]. Cholesterol efflux leads to changes in membrane architecture and fluidity that gives rise to the capacitation of the frozen sperm cells.…”

Section: Cholesterol-phospholipids Content and C/p Ratio Of Spermatozoamentioning

confidence: 99%

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Cholesterol loaded cyclodextrin increases freezability of buffalo bull (Bubalus bubalis) spermatozoa by increasing cholesterol to phospholipid ratio

Rajoriya¹,

Prasad²,

Ghosh³

et al. 2014

Vet World

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Aim: The study was conducted to investigate the effect of cholesterol loaded cyclodextrin (CLC) on freezability of buffalo spermatozoa. Materials and Methods:Murrah buffalo bull semen samples with progressive motility of 70% and greater were used. After the evaluation of motility and livability, four equal fractions of semen samples were made. Group I was kept as control and diluted with Tris, whereas Group II, III and IV were treated with CLC solution at the rate of 2.0, 3.0 and 4.0 mg/ml respectively to obtain 120 × 10 6 sperm/ml as final spermatozoa concentration. The aliquots of all the groups were incubated for action of CLC, followed by dilution and freezing. Evaluation at pre-freeze and post-thaw stage of progressive motility, viability and level of cholesterol and phospholipid was done. Results:The mean cholesterol content (μg/100 × 10 6 spermatozoa) of Group I, II, III and IV at pre-freeze stage was 21.55±0.63, 49.56±1.38, 55.67±0.45 and 47.79±1.01 and at post-thaw stage were 13.18±0.45, 34.27±0.71, 36.21±0.48 and 33.68±0.56, respectively. At pre-freeze stage, cholesterol content was significantly (p<0.01) higher in Group III in comparison to other groups. The mean cholesterol and phospholipids content of fresh sperm was 24.14±0.58 and 51.13±0.66 μg/100 × 10 6 sperm cells, respectively, and C/P ratio of spermatozoa at fresh stage was 0.47±0.067. Conclusion:CLC treatment maintains the C/P ratio and plays an important role in maintaining membrane architecture of spermatozoa. Hence, addition of CLC may be helpful in increasing freezability of buffalo spermatozoa by increasing the C/P ratio of spermatozoa.

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Section: Cholesterol-phospholipids Content and C/p Ratio Of Spermatozoamentioning

confidence: 99%

“…The integrity of the plasma membrane is important for the spermatozoa to withstand harmful effects of the cryopreservation process. Capacitation can be reduced by adding cholesterol or cholesterol analogs to the medium [5], and can be stimulated by cholesterol acceptors such as β-cyclodextrins [6].…”

Section: Introductionmentioning

confidence: 99%

Cholesterol loaded cyclodextrin increases freezability of buffalo bull (Bubalus bubalis) spermatozoa by increasing cholesterol to phospholipid ratio

Rajoriya¹,

Prasad²,

Ghosh³

et al. 2014

Vet World

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“…Indeed, during the transition phase between liquid and crystalline states, the loss of membrane lipids can lead to membrane rigidity and impermeability to extracellular molecules [3]. Additionally, it was previously stated that the enrichment of cholesterol in membrane spermatozoa could delay equine sperm capacitation and improves the fusion between the spermatozoa and the ovule [24] and pregnancy rates [33] of mares. However for bovine, Purdy and Graham [28] showed that the addition of cholesterol in semen did not significantly affect the in vivo and in vitro fertility.…”

Section: Discussionmentioning

confidence: 99%

Substitution of egg yolk by a cyclodextrin-cholesterol complex allows a reduction of the glycerol concentration into the freezing medium of equine sperm

Blommaert¹,

Franck

Donnay

et al. 2016

Cryobiology

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“…Despite the improved freezability of bull semen (MOCÉ; GRAHAM, 2006), the fertility rates were not different between the group in which cholesterol was added to the semen and the control group (PURDY; GRAHAM, 2004). Spizziri et al (2010), observed the improvement of progressive motility of equine semen when cholesterol combined with cyclodextrin was added to the centrifugation extender; however, Oliveira et al (2010) did not observe an improvement in motility after thawing when cholesterol was added to the semen of stallions. In the present study, the addition of a plant substance equivalent of cholesterol, alone or combined with OLA, did not result in significantly different values of motility and vigor in the treatments of Experiments 1 and 3.…”

Section: Discussionmentioning

confidence: 99%

Effect of oleic-linoleic acid and ?-sitosterol to freezing extender of bulls and stallions semen

et al. 2015

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Addition of polyunsaturated fatty acids and/or cholesterol to a freezing diluent can modify the sperm plasma membrane composition, influencing its behavior during cryopreservation, thus, favoring seminal cryoresistance. The present study aimed to evaluate the effects of the addition of oleiclinoleic acid, (OLA); β-sitosterol (β-sit), a plant analog of cholesterol; and OLA + β-sit in combination to a freezing diluent, on the cryopreservation bull and stallion semen. The following variables were analyzed: motility/vigor, plasma and acrosomal membrane integrity (by Trypan Blue/Giemsa staining), mitochondrial activity (by DAB staining), and lipid peroxidation (by a TBARS assays). The lipids were added according to experimental treatments: C -control group, A1 and A2 -OLA at concentrations of 37 μM and 74 μM, B1 and B2 -β-sit at concentrations of 1 µg mL -1 and 2 µg mL -1 ; AB1 and AB2 -OLA 37 μM + β-sit 1 µg mL -1 and OLA 74 μM + β-sit 2 µg mL -1 , respectively. The study was divided into three experiments; in Experiment 1, the concentrations of the groups A1, B1, and AB1 were evaluated, whereas in Experiment 2 the concentrations of the groups A2, B2, and AB2 were analyzed, both experiments were performed with bull semen. We conducted Experiment 3 using equine semen with the addition of lipids at all of the concentrations described. Data were subjected to analysis of variance, using the GLM procedure of SAS, with treatment means compared by Duncan test considering 5% significance. These variables differed significantly after thawing the semen post-collection. However, there was no significant difference between treatments when variables were compared within the same time point, except for Experiment 2, where there was a decrease in motility and vigor decrease postthaw in the groups following β-sit addition (C -51.0 ± 13.7%/2.9 ± 0.4; B2 -35.8 ± 15.8%/2.3 ± 0.6; AB2 -38.5 ± 16.6%/2.5 ± 0.5, respectively; p < 0.05). In conclusion, the tested concentrations of these lipids did not confer greater cryoresistance to the spermatozoa, and were not effective in preserving the structural integrity of plasma and acrosomal membranes after thawing. Furthermore, there was no change in the mitochondrial activity and lipid peroxidation due to lipids addition. Key words: Cholesterol, cryopreservation, PUFAs, spermatozoa ResumoA adição de ácidos graxos poli-insaturados e/ou colesterol ao diluidor pode modificar a composição da membrana plasmática do espermatozoide, influenciando seu comportamento frente à criopreservação e favorecendo, assim, a crioresistência seminal. O presente trabalho teve como objetivo avaliar o efeito da adição do ácido oleico-linoleico (AOL), β-sitosterol (β-sit), um análogo vegetal do colesterol, e da associação AOL + β-sit, ao diluente de congelação de sêmen bovino e equino. As variáveis analisadas foram motilidade/ vigor, integridade das membranas plasmática e acrossomais (Trypan Blue/Giemsa), atividade mitocondrial (DAB) e nível de peroxidação lipídica (TBARS). Os lipídios foram adicionados de acor...

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Cholesterol addition protects membrane intactness during cryopreservation of stallion sperm

Cited by 41 publications

References 29 publications

Cholesterol loaded cyclodextrin increases freezability of buffalo bull (Bubalus bubalis) spermatozoa by increasing cholesterol to phospholipid ratio

Cholesterol loaded cyclodextrin increases freezability of buffalo bull (Bubalus bubalis) spermatozoa by increasing cholesterol to phospholipid ratio

Substitution of egg yolk by a cyclodextrin-cholesterol complex allows a reduction of the glycerol concentration into the freezing medium of equine sperm

Effect of oleic-linoleic acid and ?-sitosterol to freezing extender of bulls and stallions semen

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