Effect of Catalase and Superoxide Dismutase on Motility, Viability and Acrosomal Integrity of Frozen‐Thawed Cat Spermatozoa

Thiangtum, Khongsak; Pinyopummin, Anuchai; Hori, Tatsuya; Kawakami, Eiichi; Tsutsui, Toshihiko

doi:10.1111/j.1439-0531.2009.01420.x

Cited by 22 publications

(19 citation statements)

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“…In domestic cats, reduction on acrosomal integrity following sperm cryopreservation is a common finding [22,23,25]. Furthermore, although the inclusion of antioxidants to freezing extenders showed to be unable to minimize acrosome injury in cat spermatozoa [26][27][28], the addition of detergents such as Equex STM paste and OEP remains controversial in this matter [22,24,25].…”

Section: Discussionmentioning

confidence: 97%

“…In domestic cats, besides one report addressing the use of DMSO [20], all studies on sperm-freezing protocols have utilized glycerol as their cryoprotectant and in a final concentration ranging from 3% [11,21] to 7% [5,6,22], with 4% [7,9,10,23,24] and 5% [25][26][27][28] being the most commonly used. Regarding optimal final glycerol concentration, because different extenders and freezing/thawing methods were used in most studies, a comparison among them is very complicated and offers reduced reliability.…”

Section: Introductionmentioning

confidence: 99%

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Cryoprotective effect of different glycerol concentrations on domestic cat spermatozoa

et al. 2013

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Cryopreservation of spermatozoa is a pivotal tool in assisted reproduction, and studies aiming to establish optimal freezing/thawing protocols are essential to enhance sperm survival. The objectives of the present study were to (1) compare the cryoprotective efficiency of three different glycerol concentrations (3%, 5%, and 7%) on the basis of post-thaw sperm quality and (2) investigate whether the incidence of morphologically abnormal sperm in fresh samples is related to cryodamage sensitivity. Semen was collected from six tomcats using an artificial vagina (total 18 ejaculates). Each ejaculate was diluted using Tris-egg yolk-based extender (TEY), evaluated, equally divided into three aliquots, and rediluted using TEY with and without glycerol to achieve final concentrations of 3%, 5%, and 7%. Samples were loaded into 0.25 mL straws, equilibrated for 60 minutes at 5 °C, frozen, and then thawed at 46 °C for 12 seconds. Fresh and frozen-thawed samples were evaluated for sperm motion parameters (computer-assisted sperm analysis), plasma membrane integrity (PMI; propidium iodide and carboxyfluorescein diacetate), and DNA integrity (acridine orange). Plasma and acrosomal membrane integrity were assessed by flow cytometry (propidium iodide and fluorescein isothiocyanate-conjugated pea (Pisum sativum) agglutinin) immediately after thawing. Sperm motion parameters were also evaluated at 30 and 60 minutes of postincubation. For all treatment groups, cryopreservation significantly impaired the PMI and sperm motion parameters, except for straightness and amplitude of lateral head displacement. DNA integrity showed a slight reduction (P < 0.05) when 3% glycerol was used. The percentage of total motility, progressive motility, and rapid spermatozoa were significantly lower immediately after thawing and up to 60 minutes of incubation for the 3% glycerol group when compared with 5% and 7%. No difference (P > 0.05) was found for PMI, acrosome integrity, and DNA integrity among post-thaw groups. However, higher (P < 0.05) incidence of viable cells with reacted acrosome and dead cells with intact acrosome were observed with 7% and 3% glycerol, respectively. Percentage of morphologically abnormal spermatozoa in fresh sample was positively correlated with PMI only in the 3% glycerol group and negatively correlated with sperm motility in the 5% and 7% groups. In conclusion, the final concentration of 5% glycerol offered better cryoprotective effect for ejaculated cat sperm, and the relationship found between prefreezing sperm morphology and post-thaw sperm quality showed to be dependent on final glycerol concentration.

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Section: Discussionmentioning

confidence: 97%

Section: Introductionmentioning

confidence: 99%

Cryoprotective effect of different glycerol concentrations on domestic cat spermatozoa

et al. 2013

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“…Catalase, a natural antioxidant, improves antioxidant status and counters free radical damages to sperm plasma membrane. This improves motility, viability and functional integrity of spermatozoa (Thiangtum et al. 2009; Paudel et al.…”

Section: Introductionmentioning

confidence: 99%

Effect of Incorporation of Additives in Tris‐Based Egg Yolk Extender on Buffalo (Bubalus bubalis) Sperm Tyrosine Phosphorylation During Cryopreservation

Kumar

Atreja

2011

Reprod Domestic Animals

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Phosphorylation of tyrosine residues on sperm protein is a known indicator of capacitation and a major intracellular signalling event. There is evidence that sperm cryopreservation promotes tyrosine phosphorylation and is associated with reduced fertility of spermatozoa. Under this study, cryoprotective role of different additives namely taurine, trehalose, catalase and 4-bromophenacyl bromide on buffalo sperm quality was evaluated. Buffalo semen was cryopreserved in tris-based egg yolk extender supplemented with additives like taurine (50 mm) or trehalose (100 mm) or 4-bromophenacyl bromide (200 μm) or catalase (100 U/ml) and used for assessment of levels of tyrosine phosphorylation in frozen-thawed spermatozoa. The results obtained were compared with the level of protein tyrosine phosphorylation of semen cryopreserved in tris-based egg yolk extender without additives. Proteins were extracted from a total number of nine ejaculates from three individual buffalo bulls chosen at random and analysed for tyrosine phospho-proteins using SDS-PAGE followed by immunoblotting. Monoclonal anti-phosphotyrosine antibody (Clone pT-154) was used as primary antibody followed by treatment with HRP-conjugated secondary antibody. Signals were detected on X-ray film using chemiluminescence. Nine proteins (p20, p30, p32, p38, p49, p56, p59, p72 and p86) were found to be tyrosine phosphorylated in cryopreserved spermatozoa. Supplementation of additives significantly (p<0.05) reduced the level of protein tyrosine phosphorylation in spermatozoa. Moreover, this study showed improved (p<0.05) post-thaw motility, viability and membrane integrity of spermatozoa on addition of these additives. The results obtained clearly indicate reduced level of capacitation like changes on supplementation of additives in terms of protein tyrosine phosphorylation.

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“…The extended semen was packaged in 0.25 ml plastic straws and sealed with heat pins. The straws were then cryopreserved using a one-step freezing method described by Thiangtum et al (2009). Briefly, the straws were stored horizontally on a metal rack in a Styrofoam box, 7 cm above the liquid nitrogen surface, for 10 min and then plunged into liquid nitrogen.…”

Section: Methodsmentioning

confidence: 99%

“…However, freeze/thawing process reduces sperm survival and fertilizing ability of cat spermatozoa (Luvoni, 2006;Thiangtum et al, 2009;Terrell et al, 2012), being sperm motility the most sensitive sperm parameter altered due to cryo-damage (Filliers et al, 2008).…”

Section: Introductionmentioning

confidence: 99%

Effect of Sperm Selection Techniques in Frozen/Thawed Cat Spermatozoa on Sperm Motility Analyzed by CASA System

2017

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CHEUQUEMÁN, C.; SÁNCHEZ, R. & RISOPATRÓN, J.Effect of sperm selection techniques in frozen/thawed cat spermatozoa on sperm motility analyzed by CASA system. Int. J. Morphol., 35(4):1495Morphol., 35(4): -1501Morphol., 35(4): , 2017. SUMMARY:Freeze/thawing process reduces sperm survival and fertilizing ability of cat spermatozoa, with sperm motility being the most sensitive sperm parameter altered, due to cryo-damage. In this context, swim-up and density gradient processing methods can help to recover high motile and normal spermatozoa. Maximizing the use of frozen semen sample is essential, especially in endangered felids or high value cats in which sample size, number of samples or access to semen collection is reduced. To our knowledge, there is no previous report describing an in depth analysis of sperm motility improvement, after sperm selection techniques in frozen cat semen. Accordingly, we evaluated the effect of percoll gradient (PG) and swim up (SU) sperm selection techniques on sperm motility parameters and sperm recovery rate in frozen/thawed spermatozoa of domestic cat. Next, we evaluated the individual effect of the cat over sperm motility after PG sperm selection of frozen/thawed spermatozoa. SU and PG improved significantly all sperm motility parameters of frozen/thawed cat spermatozoa compared to simple washing. However, PG allows better sperm recovery from the original frozen sample and works mostly homogeneously among individual cats. This new information could help to maximize the use of frozen semen in endangered felids or high value domestic cats for its subsequent application on in vitro fertilization and artificial insemination.

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Effect of Catalase and Superoxide Dismutase on Motility, Viability and Acrosomal Integrity of Frozen‐Thawed Cat Spermatozoa

Cited by 22 publications

References 19 publications

Cryoprotective effect of different glycerol concentrations on domestic cat spermatozoa

Cryoprotective effect of different glycerol concentrations on domestic cat spermatozoa

Effect of Incorporation of Additives in Tris‐Based Egg Yolk Extender on Buffalo (Bubalus bubalis) Sperm Tyrosine Phosphorylation During Cryopreservation

Effect of Sperm Selection Techniques in Frozen/Thawed Cat Spermatozoa on Sperm Motility Analyzed by CASA System

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