Enriching membrane cholesterol improves stability and cryosurvival of buffalo spermatozoa

Rajoriya, J. S.; Prasad, J.K.; Ramteke, Suman; Perumal, P.; Ghosh, Subrata; Singh, Mamta; Pande, Megha; Srivastava, N.

doi:10.1016/j.anireprosci.2015.11.014

Cited by 27 publications

(23 citation statements)

References 44 publications

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“…Our study revealed loss of cholesterol during cryopreservation. This was in agreement with the reports of Rajoriya et al (2016), who reported reduced cholesterol content in frozen-thawed sperm as compared to the prefreeze sperm.…”

Section: Discussionsupporting

confidence: 93%

Effect of various levels of dissolved oxygen on reactive oxygen species and cryocapacitation‐like changes in bull sperm

Amin

Prasad

Ghosh

et al. 2018

Reprod Domestic Animals

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The present investigation was carried out to study the effect of various levels of dissolved oxygen (DO) on reactive oxygen species (ROS) and cryocapacitation-like changes in bull sperm. Egg yolk-Tris-glycerol (EYTG) extender was split into four subextenders; viz., Extender I (control; no flushing with liquid nitrogen (LN )), Extender II, Extender III and Extender IV were flushed with LN for 40, 16 and 8 min, respectively. The DO levels were standardized to 11.7, 2, 4 and 8 ppm, respectively, in control (Extender I), Extender II, Extender III and Extender IV. Ejaculates with mass motility of ≥ 3+ were divided into group I (diluted with Extender I), group II (diluted with Extender II), group III (diluted with Extender III) and group IV (diluted with Extender IV) up to 80 × 10 sperm/ml. Extended semen samples were packed in French mini straws (0.25 ml), equilibrated and cryopreserved. Semen samples were evaluated at prefreeze and post-thaw stage for various parameters (DO, progressive motility (PM), viability (VIB), acrosomal integrity (AI), hypo-osmotic swelling (HOS) test, ROS, cholesterol (C) and phospholipid (P). The percentage of PM, VIB, AI, HOS test, cholesterol (C) and phospholipid (P) levels, and capacitated sperm were significantly (p < 0.05) higher in groups III and IV as compared to groups I and II. However, the acrosome-reacted sperm (%; pattern AR) were significantly (p < 0.05) decreased in group III as compared to all other groups. Besides the proportion of sperm displaying tyrosine-phosphorylated pattern, EA (fluorescence at both equatorial and anterior acrosomal regions, i.e. high capacitation level) was significantly (p < 0.05) reduced in group III compared to all other groups. In conclusion, varying DO levels in the extender significantly affect sperm quality, ROS production and capacitation-like changes in bulls.

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Section: Discussionsupporting

confidence: 93%

Effect of various levels of dissolved oxygen on reactive oxygen species and cryocapacitation‐like changes in bull sperm

Amin

Prasad

Ghosh

et al. 2018

Reprod Domestic Animals

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“…Cryoprotective actions of CLC on cryocapacitation and improvement of sperm integrity have been reported (Longobardi et al, 2017). In our previous study also, semen quality profiles were significantly higher in CLC treated than in control and of which, 3 mg CLC had shown significantly higher value than in other treatment groups (3 vs. 1, 2 and 4 mg/120 × 10 6 spermatozoa of CLC) (Rajoriya et al, 2014, 2016). However, studies on effect of CLC on prevention of cryocapacitation‐like changes, membrane fluidity, and on improvement of in vitro fertility was lacking in bubaline species.…”

Section: Discussionmentioning

confidence: 68%

“…The most interesting results arising from this study was that there was a strong reduction in sperm cryocapacitation when the buffalo sperm was processed with CLC. This is particularly important because the percentage of sperm undergoing cryocapacitation in buffalo sperm is much higher than in other domestic species (Elkhawagah et al, 2014) because the buffalo spermatozoon contains poor C: P ratio (0.34, 0.24 and 0.17 at fresh, pre‐freeze and post‐thaw, respectively, Rajoriya et al, 2014; Rajoriya et al, 2016) and easily susceptible to cold shock. These results clearly indicated that the Gr III (3 mg CLC) followed by Gr II (2 mg CLC) has protected the sperm cells from cryocapacitation and premature acrosomal reaction in buffalo semen.…”

Section: Discussionmentioning

confidence: 99%

Exogenous cholesterol prevents cryocapacitation‐like changes, membrane fluidity, and enhances in vitro fertility in bubaline spermatozoa

Rajoriya

Prasad

Ramteke

et al. 2020

Reprod Domestic Animals

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A study was conducted to determine the optimum dosage of the exogenous cholesterol‐loaded cyclodextrins (CLC) to get maximum cryoprotection for bubaline spermatozoa. In the present study, 120 × 106 spermatozoa were incubated in 2, 3 and 4 mg of CLC as grouped as Gr II, III and IV, respectively, and sperm progressive motility, intracellular Ca2+, capacitation status by protein tyrosine phosphorylation (PTP) assay and zona binding per cent (ZBP) and cleavage rate (CR) of the cryopreserved buffalo spermatozoa by in vitro fertility assay were assessed in comparison with an untreated control group (Gr I). Results revealed that there was a significant (p < .05) linear decrease in percentage of sperm population with higher intracellular Ca2+ and percentage of sperm population with medium or high capacitated by PTP in CLC treated from 2 to 3 mg and then increased to 4 mg/120 × 106 spermatozoa whereas sperm progressive motility, percentage of sperm population with low capacitated, ZBP and CR were increased significantly (p < .05) in sperm population treated from 2 to 3 mg CLC and then decreased to 4 mg/120 × 106 spermatozoa. The study has clearly indicated that CLC at 3 mg/120 × 106 spermatozoa has maximum beneficial effects in protection of sperm progressive motility, membrane fluidity (low intracellular Ca2+); prevention of cryocapacitation (low capacitation pattern in immunolocalization) and enhancement of in vitro ZBP and CR. Post‐thaw motility of the CLC‐treated sperm has shown positively significant (p < .05) correlation with sperm population with low intracellular Ca2+, low capacitated sperm population, ZBP and CR, whereas it was negatively (p < .05) correlated with sperm population with high intracellular Ca2+, medium or high capacitated sperm. The present study has revealed for the first time that incubation of spermatozoa with CLC of higher dose (>3 mg/120 × 106 spermatozoa) had adverse effects on sperm cryopreservation, although incubation of sperm with 3 mg/120 million prior to processing had minimised the freezing–thawing‐associated damages in bubaline species.

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“…The cholesterol decrease in this membrane also plays a role in the induction of the acrosome reaction. The cholesterol/phospholipid ratio of the sperm membrane is a major determinant in membrane fluidity and stability during cryopreservation [ 22 ]. Cholesterol reduces the transition temperature of membrane, and maintains them in a fluid state at reduced temperatures, thereby, reducing the membrane damage that occurs at low temperatures [ 23 ].…”

Section: Discussionmentioning

confidence: 99%

Effect of methyl-beta-cyclodextrin on the viability and acrosome damage of sex-sorted sperm in frozen-thawed bovine semen

Lee

et al. 2016

J of Biol Res-Thessaloniki

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BackgroundThe regulation of methyl-beta-cyclodextrin (MBCD) on cryodamage on X- and Y-sperm during cryopreservation of semen was investigated. The semen was collected from ten healthy bulls of proven fertility by an artificial vagina. The bovine sperm treated with MBCD fresh solution (0, 1, 5, 10, and 20 mM). The sperms were evaluated for viability and acrosome damage using flow cytometry. Moreover, X- and Y-sperm in frozen-thawed bovine semen were sorted by flow cytometry after Hoechst 33342-dyed, and the viability and acrosome damage of sperms were analyzed.ResultsSperm viability in frozen-thawed semen was decreased by MBCD (p < 0.05), also the acrosome damage of sperm was significantly increased (p < 0.05). Moreover, we sorted X- and Y-sperm from frozen-thawed bovine semen for observing the viability and acrosome damage on the separated X- and Y-sperm after MBCD treatment. Viability of X-sperm was significantly lower than that of Y-sperm (p < 0.05). Also, acrosome damage of X-sperm was significantly higher than Y-sperm (p < 0.05).ConclusionsMethyl-beta-cyclodextrin enhances damage of sperm in frozen-thawed bovine semen, and X-sperm is more sensitive than Y-sperm in cell damage. These results demonstrate that MBCD can inhibit viability of spermatozoa in frozen-thawed bovine semen (for X-sperm, especially).

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Enriching membrane cholesterol improves stability and cryosurvival of buffalo spermatozoa

Cited by 27 publications

References 44 publications

Effect of various levels of dissolved oxygen on reactive oxygen species and cryocapacitation‐like changes in bull sperm

Effect of various levels of dissolved oxygen on reactive oxygen species and cryocapacitation‐like changes in bull sperm

Exogenous cholesterol prevents cryocapacitation‐like changes, membrane fluidity, and enhances in vitro fertility in bubaline spermatozoa

Effect of methyl-beta-cyclodextrin on the viability and acrosome damage of sex-sorted sperm in frozen-thawed bovine semen

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