Effect of commercial long-term extenders on metabolic activity and membrane integrity of boar spermatozoa stored at 17°C

Dziekońska, Anna; Fraser, Leyland; Majewska, Anna; Lecewicz, Marek; Zasiadczyk, Łukasz; Kordan, Władysław

doi:10.2478/pjvs-2013-0072

Cited by 14 publications

(14 citation statements)

References 27 publications

(66 reference statements)

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“…With the method reported here consistently 10- to 20-fold higher ATP concentrations in boar spermatozoa were measured compared to previous reports (Long and Guthrie, 2006; Dziekońska and Strzeżek, 2011; Dziekońska et al, 2013). A key feature of the revised assay is an improved method for ATP extraction by phosphatase inhibition on ice and use of a boiling buffer.…”

Section: Discussionsupporting

confidence: 43%

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Energy metabolic state in hypothermically stored boar spermatozoa using a revised protocol for efficient ATP extraction

et al. 2016

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Mammalian spermatozoa utilize ATP as the energy source for key functions on the route to fertilization. ATP and its precursor nucleotides ADP and AMP are regularly investigated in sperm physiology studies, mostly by bioluminescence assays. Assay results vary widely, mainly due to different efficiencies in nucleotide extraction and prevention of their enzymatic degradation. Here, we describe a revised, validated protocol for efficient phosphatase inhibition and adenine nucleotide extraction resulting in consistently high ATP concentrations exceeding previously reported values for boar spermatozoa up to 20-fold. The revised assay is applicable for determining ATP concentrations and adenylate energy charge in extracts from fresh and frozen samples, thereby allowing simultaneous assessment of semen samples from long-term storage experiments. After validation, the assay was applied to liquid-preserved boar spermatozoa stored at 17°C and 5°C for 24 and 72 h. Cooling to 5°C, but not storage duration, reduced ATP concentration in spermatozoa (P<0.05), which was accompanied by the appearance of AMP and ADP in the preservation medium. ATP and energy charge were highly correlated to the proportion of membrane-intact spermatozoa, supporting the idea of nucleotides leaking through disrupted membranes in cold-shocked cells. The present assay allows highly standardized studies of energy metabolism in spermatozoa.

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Section: Discussionsupporting

confidence: 43%

“…Studies on fresh and stored boar semen using variants of the bioluminescence method report variable ATP contents ranging between 5 and 152 pmol/10 6 sperm (Long and Guthrie, 2006; Dziekońska and Strzeżek, 2011; Dziekońska et al, 2013). Ideally, an ATP assay should allow detection of all intracellular ATP.…”

Section: Introductionmentioning

confidence: 99%

Energy metabolic state in hypothermically stored boar spermatozoa using a revised protocol for efficient ATP extraction

et al. 2016

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“…The situation is not clear since other analysed parameters of spermatozoa quality (PMI, MMP, ATP content, GPx activity) were highest in Androhep. The obtained results remain partially different from those previously obtained (Dubé et al 2004, Dziekońska et al 2013, which demonstrated that the AH, as compared with the BTS, helped maintain better motility and the percentage of spermatozoa with progressive motion, as well as the integrity of membranes in the stored semen. However, the present study lacks statistically significant differences between the extenders in the analysis of sperm motility, in spite of the considerable difference between the values, which could indicate the need for more replications.…”

Section: Discussioncontrasting

confidence: 56%

Effects of storage in different semen extenders on the pre-freezing and post-thawing quality of boar spermatozoa

Dziekońska¹,

Zasiadczyk²,

Lecewicz³

et al. 2015

Polish Journal of Veterinary Sciences

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The aim of this study was to investigate the effects of storage of semen in different commercial extenders on the pre-freezing and post-thawing quality of boar spermatozoa. Semen was diluted in BTS, Androhep (AH) and Gedil (GD), stored for 24 h at 17 o C, and then frozen in accordance with the cryopreservation protocol. Analyses of the quality of spermatozoa included: motility, normal apical ridge (NAR) acrosome, plasma membrane integrity (PMI), mitochondrial membrane potential (MMP), measurements of ATP content and activity of superoxidase dismutase (SOD) and glutathione peroxidase (GPx). Prior to the freezing process, no significant effect of the extender on the sperm quality parameters was noted. After thawing the spermatozoa it was demonstrated that the type of extender used influenced PMI, MMP, ATP content and activity of GPx. In the AH extender the percentage of spermatozoa with PMI and ATP content in spermatozoa was significantly higher (P<0.05) as compared to the BTS or GD extenders. In addition, semen stored in the AH was characterised by a statistically higher (P<0.05) percentage of spermatozoa with MMP and increased activity of GPx as compared with the BTS. The results obtained indicate that for the cryopreservation process, boar spermatozoa stored for 24 hours in liquid state can be used. However, the type of extender used prior to freezing may have a significant effect on the post-thawing quality of the spermatozoa. The AH extender better secured the quality of thawed boar spermatozoa as compared with the BTS or GD.

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“…In the present study, semen stored at 16 °C failed to meet the 60% threshold of spermatozoa with progressive motility after the sixth day, while in the semen preserved at 5 °C and 25 °C this happened on the second day. The particular extenders used in porcine AI may have different effects on motility and other features of sperm quality (Johnson et al, 2000;Fantinati et al, 2009;Martín-Hidalgo et al, 2013a;Dziekońska et al, 2013). A drop in motility below 60% could, therefore, occur at different points in time (Martin-Hidalgo et al, 2011;Dziekońska et al, 2013) and under various semen storage temperatures (Paulenz et al, 2000).…”

Section: The Influence Of Temperature and Time Of Semen Storage On Thmentioning

confidence: 99%

Storage temperature of boar semen and its relationship to changes in sperm plasma membrane integrity, mitochondrial membrane potential, and oxidoreductive capability

Gączarzewicz¹,

Udała²,

Piasecka³

et al. 2015

Turk J Biol

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The aim of this study was to analyze changes in sperm plasma membrane integrity, mitochondrial activity, and extracellular environment during storage of boar semen at 5 °C, 16 °C, and 25 °C for 10 days. Progressive sperm motility, plasma membrane integrity (SYBR-14/PI-test and HOS test), aspartate aminotransferase (AAT), and mitochondrial activity (JC-1-test and NADH-dependent NBT assay), as well as pH and osmolality were assessed in nine ejaculates of Polish Landrace boars. The plasma membrane integrity of the semen stored at 5 °C was similar to that of the semen stored at 16 °C and 25 °C; however, an increase in AAT activity of the semen stored at 5 °C revealed sperm membrane disorders as early as day 2. Significant differences in the progressive motility of the semen stored at 5 °C and 16 °C were observed at each of the evaluation times. This reduced motility was consistent with decreased sperm mitochondrial transmembrane potential and oxidoreductive capability. We inferred that the cooling of boar semen to 5 °C increases the permeability of the sperm plasma membrane, which may not be revealed by SYBR-14/PI staining or HOS testing. The loss of boar sperm motility that occurs during hypothermic liquid preservation may be connected with alterations in the plasma membrane stability and disorders of mitochondrial transmembrane potential and oxidoreductive capability.

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Effect of commercial long-term extenders on metabolic activity and membrane integrity of boar spermatozoa stored at 17°C

Cited by 14 publications

References 27 publications

Energy metabolic state in hypothermically stored boar spermatozoa using a revised protocol for efficient ATP extraction

Energy metabolic state in hypothermically stored boar spermatozoa using a revised protocol for efficient ATP extraction

Effects of storage in different semen extenders on the pre-freezing and post-thawing quality of boar spermatozoa

Storage temperature of boar semen and its relationship to changes in sperm plasma membrane integrity, mitochondrial membrane potential, and oxidoreductive capability

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