The role of different trehalose concentrations and cooling rates in freezing of ram semen

Ongun;BUCAK, UYSAL

doi:10.1501/vetfak_0000002176

Cited by 28 publications

(1 citation statement)

References 24 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…It was prepared according to the manufacturer's specifications -excluding egg yolk -and was stored at 5°C. Then, five treatments were developed by the supplementation to the extender with CEY: chicken egg yolk at 20% v/v (Waheed et al, 2012); CCEY: centrifuged CEY at 20% v/v (Nouri et al, 2013); LDL: LDL at 8% v/v (Moussa et al, 2002); T: trehalose at 100 mM (Uysal & Bucak, 2009); and TLDL: trehalose at 100 mM plus LDL at 8% v/v. The semen samples were extended in the different treatments to a final sperm concentration of 30 × 10 6 ml −1 .…”

Section: Treatmentsmentioning

confidence: 99%

Membrane stability and mitochondrial activity of bovine sperm frozen with low‐density lipoproteins and trehalose

Varela

Rojas

Betancur

2019

Reprod Domestic Animals

View full text Add to dashboard Cite

Cryopreservation results in the destabilization of the sperm plasma membrane, leading to negative side effects such as premature cryocapacitation, apoptosis and the low mitochondrial activity of bovine spermatozoa. Low‐density lipoproteins (LDL) and trehalose have been used in seminal freezing to protect the integrity and stability of sperm membranes. Likewise, trehalose can increase the mitochondrial activity of sperm. The objective of this study was to evaluate the membrane stability and mitochondrial activity of bovine sperm after being frozen and treated with LDL sources and trehalose. Ten ejaculates from five bulls were cryopreserved under the treatments, CEY: chicken egg yolk (20% v/v); CCEY: centrifuged CEY (20% v/v); LDL: LDL (8% v/v); T: trehalose (100 mM); and TLDL: T (100 mM) plus LDL (8% v/v). After thawing, membrane stability and mitochondrial membrane potential (ΔΨM) were assessed by flow cytometry through the M‐540/Yopro‐1 and DiOC6/PI probes. The structural membrane integrity (SMI) was evaluated by fluorescence microscopy using SYBR14/PI dyes. A generalized linear model was adjusted, and the means were compared using the Tukey test. Centrifuged chicken egg yolk and LDL had a higher proportion of non‐cryocapacitated non‐apoptotic sperm (M−Y−), while CEY and T had the largest populations of cryocapacitated non‐apoptotic sperm (M+Y−) and cryocapacitated apoptotic sperm (M+Y+). Centrifuged chicken egg yolk also showed a higher proportion of sperm with high‐ΔΨM. Treatments that included egg yolk or purified LDL had a positive effect on SMI. Centrifuged chicken egg yolk has a superior cryoprotective effect on membrane stability and mitochondrial activity of bovine semen over the conventional use of CEY or the individual or simultaneous use of LDL and trehalose.

show abstract