Concentração de lactato de cálcio e tempo de incubação sobre a capacidade de adesão e penetração de espermatozoides suínos na membrana perivitelina do ovo da galinha

Corcini, Carine Dahl; Silva, Betris Elert da; Brizolara, Rosa Marani Rodrigues; Gheller, Stela Mari Meneghello; Varela, Antônio Sérgio; Bongalhardo, Denise Calisto; Lucia, Thomaz

doi:10.1590/s0103-84782012000100023

Cited by 7 publications

(6 citation statements)

References 16 publications

(7 reference statements)

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“…As similaridades entre as glicoproteínas da zona pelúcida e da membrana perivitelina (MPV) do ovo de galinha, permitem a ligação dos espermatozoides de outras espécies a esta MPV (Mugnier et al 2009) e simula a interação dos gametas (Corcini 2010) sendo, portanto, um teste eficiente para se verificar a habilidade dos espermatozoides em sofrer a reação acrossomal (Corcini et al 2012). O uso da MPV do ovo de galinha, em relação aos ovócitos, é muito mais vantajoso porque permite, sobre diferentes fragmentos de uma mesma membrana, analisar diferentes partidas de sêmen, minimizando o efeito fêmea na análise e também por ser menos dispendioso que o ovócito (Csermak Junior 2011).…”

Section: Introductionunclassified

Características do sêmen a fresco e descongelado de garanhões da raça Nordestina

et al. 2015

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RESUMO.-Este estudo descreveu as características seminais, da membrana plasmática e do acrossoma de espermatozoide congelado/descongelado de 19 ejaculados de garanhões da raça Nordestina. Os aspectos analisados incluíram os parâmetros físicos do sêmen fresco; a motilidade e a longevidade do sêmen diluído e descongelado; a morfologia espermática, integridade funcional e estrutural da membrana plasmática do espermatozoide e a habilidade de ligação do espermatozoide à membrana perivitelina da gema do ovo de galinha do sêmen descongelado. This paper describes the seminal characteristics of the plasma membrane of frozenthawed sperm. Nineteen ejaculates of Nordestino horse breed. The aspects analyzed in the physical parameters of fresh semen were total and progressive motility and your longevity after dilution or thawed; sperm morphology, functional and structural integrity of the plasma membrane of the sperm and the sperm-binding ability to the perivitelline membrane of the yolk (MPV) after thawed. The variables were assessed by ANOVA with post hoc test of Student Newman-Keuls test (P<0.05). The total and progressive motility were higher in diluted semen than thawed (P<0.05). The average percentage of the major, minor and total defects was lower than the limit recommended by the CBRA. The percentage of reactive to hypo-osmotic swelling test was 14.21±1.12%, the intact membrane detected by supravitally test was 62.22±9.06% and the SYBR-14 was 81.47±26.9. The ability of sperm to bind to the MPV after thawing semen was 230.39±57.09. The total and progressive motility at time 0 min of termo resistance test was higher than to 150 minutes (P<0.05), and no difference was observed in the times 10 and 30 minutes. The results demonstrate that the use of additional laboratory tests help in the process of evaluation of samples, making possible to obtain more reliable and accurate information. Although cryopreservation has caused decrease in sperm motility and was used diluents with amides to diluted and cryopreservation protocol and this minimized the osmotic damage to sperm cells and maintained the morphological, functional and structural integrity of the plasma membrane of the sperm. These results are a reference for future studies since there are no comparative data on this breed.

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Section: Introductionunclassified

Características do sêmen a fresco e descongelado de garanhões da raça Nordestina

et al. 2015

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“…() and Corcini et al. (). However, sperm populations with low binding potential most likely have a smaller number of bound sperm regardless of their cellular traits.…”

Section: Discussionmentioning

confidence: 97%

“…The relatively broad range of fertility results in the case of animals with high binding potential may occur because the maximum fertilization potential requires traits other than motility and binding according to Barbato et al (1998) and Corcini et al (2012). However, sperm populations with low binding potential most likely have a smaller number of bound sperm regardless of their cellular traits.…”

Section: Discussionmentioning

confidence: 99%

Cryopreservation of Piau‐Breed Wild Boar Sperm: Assessment of Cooling Curves and Centrifugation Regimes

Shiomi

Pinho

Lima

et al. 2015

Reprod Domestic Animals

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This study aimed to assess the effects of different cooling curves and centrifugation regimes used in cryopreservation protocols on the post-thaw viability of Piau-breed wild boar (Sus scrofa) sperm using in vitro assessment tests. Two centrifugations (800 g for 10 min and 2400 g for 3 min) and two cooling curves (conventional cooling using nitrogen vapour - freezing 1 and automated cooling using a programmed freezing machine - freezing 2) were tested. Therefore, the treatments were divided into M3 - centrifugation at 2400 g for 3 min and freezing 2; M10 - centrifugation at 800 g for 10 min and freezing 2; R3 - centrifugation at 2400 g for 3 min and freezing 1; and R10 - centrifugation at 800 g for 10 min and freezing 1. No significant differences (p > 0.05) between treatments occurred post-thawing regarding the total sperm motility means recorded. The mean values of the different treatments were not different from each other regarding the supravital staining (SV), hypo-osmotic test (HO), sperm-egg binding assay or sperm morphology. This study showed that both the cooling curve and the centrifugation regime affected the quality of post-thaw sperm, and centrifugation for shorter times and cooling curves using automated cooling are the most suitable for minimizing sperm injury.

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“…However, to our knowledge no studies regarding the standardization and evaluation of the efficiency of sperm-binding test to the perivitelline membrane of chicken eggs have been undertaken for canine sperm, mainly for post-thaw sperm samples. Conversely, tests have been performed for domestic birds, bulls, stallions, ram, boars, mice, and humans (ROBERTSON et al, 1997;BARBATO et al, 1998;WACLAWEK et al, 1998;AMANN et al, 1999;CORCINI et al, 2012;LOSANO et al, 2015). Thus, the aim of this study was to standardize and evaluate the efficiency of the sperm-binding test to the perivitelline membrane of chicken eggs as a functional method for canine semen evaluation.…”

Section: Introductionmentioning

confidence: 99%

Sperm-binding to the perivitelline membrane of chicken egg yolk as a functional test for sperm evaluation in dogs

Brito¹,

Losano²,

Angrimani³

et al. 2017

Braz. J. Vet. Res. Anim. Sci.

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During fertilization, spermatozoa interact with the zona pellucida (ZP) through the binding between the acrosome and proteins 2 and 3 (ZP2 and ZP3). The perivitelline membrane of chicken egg yolk is homologous to the mammalian ZP3, which allows the binding of sperm of several species. The aim of this study was to standardize and evaluate the efficiency of sperm-binding to the perivitelline membrane of chicken eggs as a functional method for canine semen evaluation. For this purpose, nine post-thaw sperm samples were used, which were divided into two aliquots: the first was kept in water bath at 37ºC (live sample) and the second was submitted to cold shock to induce cellular damage (dead sample). The two aliquots were mixed on five proportions, corresponding to 0%, 25%, 50%, 75%, and 100% of viable cells, and the binding test was performed by analyzing the number of spermatozoa bonded to the perivitelline membrane by means of computerized assessment of sperm motility (CASA) or conventional microscopy. Additionally, samples were submitted to sperm motility analysis, evaluation of plasmatic and acrosomal membrane integrity, and sperm mitochondrial activity. The sperm-binding test to the perivitelline membrane of chicken egg yolk was considered a feasible sperm analysis test for both fertilizing capacity and overall sperm attributes evaluation, mainly when the analysis is performed by a conventional microscope, which expands its practicality to the majority of canine reproduction laboratories. Keywords: Sperm cryopreservation. Egg yolk. Sperm analysis. Perivitelline membrane. Canine. ResumoDurante a fecundação, os espermatozoides interagem com a zona pelúcida (ZP) por meio da ligação entre o acrossomo e as proteínas 2 e 3 (ZP2 e ZP3). A membrana perivitelínica da gema de ovo de galinhas é homóloga à ZP3 de mamíferos, possibilitando a ligação espermática de diversas espécies. Este trabalho padronizou e avaliou a eficiência do teste de ligação espermática à membrana perivitelínica da gema de ovo de galinhas como avaliação funcional do sêmen de cães. Para tal, foram utilizadas nove amostras seminais previamente criopreservadas. Cada amostra foi dividida em duas alíquotas: a primeira foi mantida em banho-maria à 37ºC (vivos) e a segunda submetida a choque térmico com o intuito de induzir dano celular (mortos). As duas alíquotas foram misturadas, correspondendo a 0, 25, 50, 75 e 100% de células viáveis. As amostras foram avaliadas quanto ao número de espermatozoides ligados à membrana perivitelínica por meio da análise computadorizada da motilidade (CASA) ou microscopia convencional. Ademais, as amostras foram avaliadas quanto à motilidade espermática, integridade das membranas acrossomal e plasmática e atividade mitocondrial espermática. O teste de ligação espermática à membrana perivitelínica de ovos de galinha foi considerado um teste de análise seminal exequível tanto para avaliar a capacidade fecundante dos espermatozoides como atributos seminais gerais, especialmente quando realizado em microscopia convenci...

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Concentração de lactato de cálcio e tempo de incubação sobre a capacidade de adesão e penetração de espermatozoides suínos na membrana perivitelina do ovo da galinha

Abstract: RESUMO Este estudo testou o efeito de diferentes concentrações de lactato de cálcio e tempos de incubação sobre a capacidade de adesão e a penetração dos espermatozoides suínos nas membranas perivitelinas (MP) de

Cited by 7 publications

References 16 publications

Características do sêmen a fresco e descongelado de garanhões da raça Nordestina

Características do sêmen a fresco e descongelado de garanhões da raça Nordestina

Cryopreservation of Piau‐Breed Wild Boar Sperm: Assessment of Cooling Curves and Centrifugation Regimes

Sperm-binding to the perivitelline membrane of chicken egg yolk as a functional test for sperm evaluation in dogs

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