Efeito da adição de glutationa peroxidase e cisteína ao diluidor de congelação do sêmen equino

Barros, L.O.; Silva, Sildivane Valcácia; Almeida, Felipe Costa; Silva, E. C. B.; Carneiro, Gustavo Ferrer; Guerra, Maria Madalena Pessoa

doi:10.1590/s0102-09352013000200019

Cited by 4 publications

(4 citation statements)

References 16 publications

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“…Alguns autores afirmam que a variabilidade na resistência do espermatozóide ao processo de criopreservação está relacionada a fatores genéticos 4 . Para equinos, vários diluentes tem sido formulados, a maioria à base de gema de ovo ou leite desnatado 5 . A gema de ovo é o mais eficiente dos crioprotetores não penetrantes na proteção do espermatozóide contra o choque térmico, mas não apresenta a mesma eficiência em todas as espécies 6 .…”

Section: Introductionunclassified

Eficiência dos diluidores Tris e Botu-crio® sobre os parâmetros seminais de garanhões das raças quarto de milha e mangalarga marchador

et al. 2014

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O principal fator a limitar o processo de criopreservação de sêmen equino está relacionado à própria espécie, por apresentar uma grande variabilidade nas características dos ejaculados pós-descongelação. O objetivo desta pesquisa foi avaliar a viabilidade espermática pós-descongelação, do sêmen de garanhões das raças Quarto de Milha e Mangalarga Marchador utilizando-se dois diluentes (Tris e Botu-crio®) na criopreservação. Para tanto, foram analisadas as características físicas do sêmen fresco, funcionalidade da membrana espermática pelo Teste Hiposmótico (HOST), motilidade total e vigor pelo Teste de Termoresistência (TTR), motilidade progressiva pelo sistema computadorizado CASA (Computer Assisted Sêmen Anlyses) e integridade da membrana espermática, do acrossoma e funcionalidade de mitocôndria por microscopia de epifluorescência. O diluente Botu-crio® preservou melhor os valores de motilidade e vigor e a integridade de membrana plasmática, pós-descongelamento. Não houve diferença significativa entre as raças quanto à motilidade total e vigor no sêmen fresco, bem como para a análise desses parâmetros no Teste de Termoresistência pós-descongelamento. Garanhões da raça Quarto de Milha apresentaram percentual maior de defeitos maiores na análise de patologia espermática.

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Section: Introductionunclassified

Eficiência dos diluidores Tris e Botu-crio® sobre os parâmetros seminais de garanhões das raças quarto de milha e mangalarga marchador

et al. 2014

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“…The addition of antioxidants such as glutathione peroxidase or cysteine to equine semen extenders improves sperm VCL and VAP after thawing (BARROS et al, 2013). In humans, Moubasher et al (2013) reported increased of the percentage of progressive motility and sperm viability in samples cryopreserved in extenders with catalase.…”

Section: Resultsmentioning

confidence: 99%

Effect of catalase, superoxide dismutase and reduced glutathione in LDL extender on ovine cryopreserved sperm viability

et al. 2015

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The aim of this study was to evaluate the motility, kinetics and membrane integrity of ovine sperm cryopreserved in extenders containing 8% LDL with enzymatic antioxidants at different concentrations. Four Santa Inês rams were used to form four pools of semen (each pool containing ejaculates from four ram, totaling four ejaculates per animal). Each seminal pool was divided into eight aliquots for the following treatments: 1) Tris-glucose-glycerol (TGG) + (16%) egg yolk (control 1); 2) TGG + 8% (w/v) LDL (control 2); 3) TGG + 8% LDL + catalase 100 U/mL; 4) TGG + 8% LDL + catalase 200 U/mL; 5) TGG + 8% LDL + superoxide dismutase 100 U/mL; 6) TGG + 8% LDL + superoxide dismutase 200 U/mL; 7) TGG + 8% LDL + reduced glutathione 5 mM; and 8) TGG + 8% LDL + reduced glutathione 10 mM. The samples were packed into 0.25 mL straws, cooled (-0.25 °C/ min), maintained at 5 °C for 2 h and then frozen (-25 °C/ min) using a TK4000 ® . Immediately after thawing (38 °C/ 30 s), sperm motility and movement characteristics were assessed by computer sperm analysis (CASA). The structural integrity of the plasma and acrosomal membranes was analyzed using fluorescent dyes. The functional integrity of membranes was assessed using a hypoosmotic swelling test. As assessed by ANOVA, significant differences (P<0.05) among treatments were only observed for VCL, VSL and VAP. For the VCL variable, the 2, 3, 4, 5, 6 and 7 extenders were similar and higher than 1 and 8 extenders, the latter being similar to each other. For the VSL variable, the 3, 4, 5, 6 and 7 extenders were similar and higher than 1, 2 and 8 extenders, the latter being similar to each other. For the VAP variable, the 3, 4 and 6 extenders were similar and higher than 1, 2, 5, 7 and 8 extenders, the latter being similar to each other. In conclusion, enzymatic antioxidants as catalase and superoxide dismutase improve the protective activity of extenders containing LDL on frozen ovine sperm. concentrações. Quatro carneiros da raça Santa Inês foram utilizados para formar quatro pools de sêmen (cada pool contendo ejaculados provenientes dos quatro carneiros, totalizando quatro ejaculados por animal). Cada pool de sêmen foi dividido em oito alíquotas para os seguintes tratamentos: 1) Trisglicose-glicerol (TGG) + (16%) gema de ovo (controle 1); 2) TGG + 8% (g/L) LDL (controle 2); 3) TGG + 8% LDL + catalase 100 U/mL; 4) TGG + 8% LDL + catalase 200 U/mL; 5) TGG + 8% LDL + superóxido dismutase 100 U/mL; 6) TGG + 8% LDL + superóxido dismutase 200 U/mL; 7) TGG + 8% LDL + glutationa reduzida 5 mM; and 8) TGG + 8% LDL + glutationa reduzida 10 mM. As amostras foram envasadas em palhetas de 0,25 mL, resfriadas (-0,25 °C/min), mantidas a 5 °C por duas horas e em seguida congeladas (-25 °C/ min) usando uma máquina de congelar TK4000 ® . Imediatamente depois da descongelação (38 °C/30 s), as amostras foram submetidas à análise computadorizada (CASA) para avaliação da motilidade e cinética. A integridade estrutural das membranas plasmática e acrossomal foi analisada utilizando corantes fluor...

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“…In the absence of CAT, as demonstrated in collared peccary (Santos et al., 2018), GPX may perhaps act in a compensatory way, protecting spermatozoa from the oxidative attack triggered by H 2 O 2 (Petrujkić et al., 2014). Thus, the supplementation of the diluent medium with GPX (1 U or 5 U) has shown positive effects on the kinetic parameters of spermatozoa subjected to freezing in horses (Barros et al., 2013), and could be an alternative to be tested in the future.…”

Section: Discussionmentioning

confidence: 99%

Catalase and superoxide dismutase supplementation to the diluent for cryopreservation of collared peccary semen

Moreira,

dos Santos,

da Silva

et al. 2024

Reprod Domestic Animals

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We evaluated the effects of supplementation with catalase (CAT) and superoxide dismutase (SOD) antioxidants, isolated or combined, on the freezability of collared peccary sperm. Semen aliquots from ten individuals were diluted in Tris‐yolk with or without the presence of antioxidants CAT (200 or 400 IU/mL) and SOD (150 or 300 IU/mL) alone or in combination (CAT: 200 IU/mL + SOD: 150 IU/mL), and then cryopreserved in liquid nitrogen (−196°C). After 1 week, samples were thawed and evaluated for kinetic parameters, membrane functionality and integrity, mitochondrial activity, chromatin integrity, sperm morphology, sperm cell binding capacity and intracellular oxidative levels. Regardless of the use of antioxidants, the treatments did not differ in any of the evaluated parameters, except for the treatment with 300 IU/mL SOD, which showed the highest percentage of static cells (69.1 ± 4.4%) when compared to the group without antioxidant (51.6 ± 3.3%). In general, all samples maintained around 50% of functional membranes, 40% of intact membranes with mitochondrial activity, more than 99% of cells with condensed chromatin, as well as more than 70% of morphologically normal cells. Regarding the binding capacity, the minimum and maximum values found ranged from 33.8 to 194 spermatozoa bound to the perivitelline membrane. Finally, regardless of the presence of antioxidants, all thawed groups showed similar levels of reactive oxygen species (ROS). In conclusion, supplementation with CAT or SOD antioxidants, at the tested concentrations, does not bring benefits to freezability, and the increase in SOD concentration resulted in negative effects on sperm motility in collared peccary semen.

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Efeito da adição de glutationa peroxidase e cisteína ao diluidor de congelação do sêmen equino

Abstract: Efeito da adição de glutationa peroxidase e cisteína ao diluidor de congelação do sêmen equino

Cited by 4 publications

References 16 publications

Eficiência dos diluidores Tris e Botu-crio® sobre os parâmetros seminais de garanhões das raças quarto de milha e mangalarga marchador

Eficiência dos diluidores Tris e Botu-crio® sobre os parâmetros seminais de garanhões das raças quarto de milha e mangalarga marchador

Effect of catalase, superoxide dismutase and reduced glutathione in LDL extender on ovine cryopreserved sperm viability

Catalase and superoxide dismutase supplementation to the diluent for cryopreservation of collared peccary semen

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