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BACKGROUND: The proteomic profile of cryopreserved in vitro produced bovine embryos is little known but can provide insights on the successful application of cryo procedures in support of animal breeding. OBJECTIVE: To identify embryonic proteins and biomarkers related to improved cryotolerance of vitrified in vitro produced bovine embryos. MATERIALS AND METHODS: Proteins were isolated from embryo pools (n = 25 embryos per replicate) and analyzed using the nanoLC - MS/MS system. Further, the UniProtKB database (Uniprot -http://www.uniprot.org/) was used for protein identification. Proteins were classified based on their molecular mass, isoelectric point, and enzymatic activity. Post-translational modification predictions and functional gene ontology analysis were performed as well. Finally, a protein-protein interaction network was created to shed light on the embryo interactome. RESULTS: Based on the MS/MS approach, 66 proteins were identified from vitrified Bos taurus embryos. The retrieved proteins were presumably annotated, which allowed a description of the qualitative and functional aspects of the embryo proteome after the vitrification process. CONCLUSION: These findings allowed us to conclude that in vitro-produced vitrified embryos expressed proteins that underlie biological processes related to reproduction, stress and lipid metabolic process, which are essential to maintain embryo viability. doi.org/10.54680/fr22410110512.

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The objectives of this study were to evaluate goat sperm sorting in continuous Percoll® density gradients and gamete freezability, in the presence or absence of phenolic antioxidants. For this, semen pools were sorted, frozen, and evaluated. The non-selected group (NSg) presented lower progressive motility (PM), linearity (LIN), straightness (STR), and wobble (WOB) than the selected groups, and straight line velocity (VSL) compared to those with catechin or resveratrol. The amplitude of lateral head displacement (ALH) was higher in NSg, and quercetin reduced the mitochondrial membrane potential (MMP). After thawing, the NSg presented lower PM than the selected groups, VSL and VAP (average path velocity) than the selected group with or without catechin, LIN and WOB than the selected with or without catechin or resveratrol, and STR than the selected with catechin. Moreover, NSg presented higher ALH and BCF than the samples selected with or without catechin. Plasma membrane integrity and intact and living cells were higher in the selected groups, and MMP was lower in the NSg and the selected group with quercetin. Thus, centrifugation in Percoll® continuous density gradients is a viable methodology to select goat sperm compatible with the freezing, especially in the presence of catechin or resveratrol.(AU)

Objetivou-se avaliar a separação de espermatozoides caprinos em gradientes de densidade contínuos de Percoll® e a congelabilidade espermática, com ou sem antioxidantes fenólicos. Para tal, pools seminais foram selecionados, congelados e avaliados. O grupo não selecionado (gNS) apresentou menor motilidade progressiva (MP), linearidade (LIN), retilinearidade (STR) e oscilação (WOB) do que os selecionados, bem como menor velocidade linear progressiva (VSL) do que os com catequina ou resveratrol. A amplitude de deslocamento lateral de cabeça (ALH) foi maior no gNS e a quercetina reduziu o potencial de membrana mitocondrial (PMM). Após a descongelação, o gNS manifestou menor MP do que os selecionados, menor VSL e VAP (velocidade média da trajetória) do que os com ou sem catequina, menor LIN e WOB do que os com ou sem catequina ou resveratrol, e menor STR do que os com catequina, além de maior ALH e BCF do que os com ou sem catequina. A integridade da membrana plasmática e as células intactas e vivas foram maiores nas amostras selecionadas e o PMM foi inferior no gNS e no selecionado com quercetina. Portanto, a centrifugação em gradientes contínuos de densidade de Percoll® é uma metodologia viável para selecionar espermatozoides caprinos compatíveis com a congelação, especialmente na presença de catequina ou resveratrol.(AU)

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RESUMEN

The objectives of this study were to evaluate goat sperm sorting in continuous Percoll® density gradients and gamete freezability, in the presence or absence of phenolic antioxidants. For this, semen pools were sorted, frozen, and evaluated. The non-selected group (NSg) presented lower progressive motility (PM), linearity (LIN), straightness (STR), and wobble (WOB) than the selected groups, and straight line velocity (VSL) compared to those with catechin or resveratrol. The amplitude of lateral head displacement (ALH) was higher in NSg, and quercetin reduced the mitochondrial membrane potential (MMP). After thawing, the NSg presented lower PM than the selected groups, VSL and VAP (average path velocity) than the selected group with or without catechin, LIN and WOB than the selected with or without catechin or resveratrol, and STR than the selected with catechin. Moreover, NSg presented higher ALH and BCF than the samples selected with or without catechin. Plasma membrane integrity and intact and living cells were higher in the selected groups, and MMP was lower in the NSg and the selected group with quercetin. Thus, centrifugation in Percoll® continuous density gradients is a viable methodology to select goat sperm compatible with the freezing, especially in the presence of catechin or resveratrol.(AU)

Objetivou-se avaliar a separação de espermatozoides caprinos em gradientes de densidade contínuos de Percoll® e a congelabilidade espermática, com ou sem antioxidantes fenólicos. Para tal, pools seminais foram selecionados, congelados e avaliados. O grupo não selecionado (gNS) apresentou menor motilidade progressiva (MP), linearidade (LIN), retilinearidade (STR) e oscilação (WOB) do que os selecionados, bem como menor velocidade linear progressiva (VSL) do que os com catequina ou resveratrol. A amplitude de deslocamento lateral de cabeça (ALH) foi maior no gNS e a quercetina reduziu o potencial de membrana mitocondrial (PMM). Após a descongelação, o gNS manifestou menor MP do que os selecionados, menor VSL e VAP (velocidade média da trajetória) do que os com ou sem catequina, menor LIN e WOB do que os com ou sem catequina ou resveratrol, e menor STR do que os com catequina, além de maior ALH e BCF do que os com ou sem catequina. A integridade da membrana plasmática e as células intactas e vivas foram maiores nas amostras selecionadas e o PMM foi inferior no gNS e no selecionado com quercetina. Portanto, a centrifugação em gradientes contínuos de densidade de Percoll® é uma metodologia viável para selecionar espermatozoides caprinos compatíveis com a congelação, especialmente na presença de catequina ou resveratrol.(AU)

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The aim of this study was to evaluate the effects of different concentrations of (+)-catechin or (-)-epigallocatechin gallate (EGCG) on goat semen freezability. Poolsof semen were processed (Experiment 1: 0, 15, 25, 50, 75, or 100µM (+)-catechin; Experiment 2: 0, 15, 25, 50, 75, or 100µM EGCG) and frozen. After thawing, the samples were evaluated for kinematics, plasma membrane (PMi) and acrosome integrity, morphology, and oxidative stress, at 0 and 1h. In Experiment 1, at 0h, VSL and VAP were greater (P<0.05) with 15µM than with 50 and 100; WOB was lower (P<0.05) with 100µM than with 0, 15, and 25; and BCF was higher (P<0.05) with 75 and 100µM than with 0. In turn, in Experiment 2, progressive motility was higher (P<0.05) with0 and 15µM than with50 and 75; LIN was lower (P<0.05) with75 and100µM than with0 and 15; WOB was higher (P<0.05) with0 and 15µM; and PMi was greater (P<0.05) with100µM than 0. Thus, (+)-catechin or EGCG at higher concentrations inhibits the kinematics of frozen goat sperm, in a transitory way, and 100µM of EGCG preserves the PMi.(AU)

Objetivou-se avaliar o efeito de diferentes concentrações de (+)-catequina ou (-)-epigalocatequina galato (EGCG) sobre a congelabilidade do sêmen caprino. Poolsseminais foram processados (experimento 1: 0, 15, 25, 50, 75 ou 100µM de (+)-catequina; experimento 2: 0, 15, 25, 50, 75 ou 100µM de EGCG) e congelados. Após a descongelação, foram avaliadas a cinética, a integridade de membrana plasmática (iMP) e acrossomal, a morfologia e o estresse oxidativo, a zero e a uma hora. No experimento 1, a zero hora, VSL e VAP foram maiores (P<0,05) com 15µM do que com 50 e100; WOB foi menor (P<0,05) com 100µM do que com 0, 15 e 25; e BCF foi maior (P<0,05) com 75 e 100µM do que com 0. No experimento 2, a motilidade progressiva foi maior (P<0,05) com 0 e 15µM do que com 50 e 75; LIN foi menor (P<0,05) com 75 e 100µM do que com 0 e 15; WOB foi maior (P<0,05) com 0 e 15µM; e iMP foi maior (P<0,05) com 100µM do que com 0. Assim, (+)-catequina ou EGCG em altas concentrações inibem, transitoriamente, a cinética de espermatozoides congelados caprinos, e 100µM de EGCG preserva a iMP.(AU)

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The aim of this study was to evaluate the effects of different concentrations of (+)-catechin or (-)-epigallocatechin gallate (EGCG) on goat semen freezability. Poolsof semen were processed (Experiment 1: 0, 15, 25, 50, 75, or 100µM (+)-catechin; Experiment 2: 0, 15, 25, 50, 75, or 100µM EGCG) and frozen. After thawing, the samples were evaluated for kinematics, plasma membrane (PMi) and acrosome integrity, morphology, and oxidative stress, at 0 and 1h. In Experiment 1, at 0h, VSL and VAP were greater (P<0.05) with 15µM than with 50 and 100; WOB was lower (P<0.05) with 100µM than with 0, 15, and 25; and BCF was higher (P<0.05) with 75 and 100µM than with 0. In turn, in Experiment 2, progressive motility was higher (P<0.05) with0 and 15µM than with50 and 75; LIN was lower (P<0.05) with75 and100µM than with0 and 15; WOB was higher (P<0.05) with0 and 15µM; and PMi was greater (P<0.05) with100µM than 0. Thus, (+)-catechin or EGCG at higher concentrations inhibits the kinematics of frozen goat sperm, in a transitory way, and 100µM of EGCG preserves the PMi.(AU)

Objetivou-se avaliar o efeito de diferentes concentrações de (+)-catequina ou (-)-epigalocatequina galato (EGCG) sobre a congelabilidade do sêmen caprino. Poolsseminais foram processados (experimento 1: 0, 15, 25, 50, 75 ou 100µM de (+)-catequina; experimento 2: 0, 15, 25, 50, 75 ou 100µM de EGCG) e congelados. Após a descongelação, foram avaliadas a cinética, a integridade de membrana plasmática (iMP) e acrossomal, a morfologia e o estresse oxidativo, a zero e a uma hora. No experimento 1, a zero hora, VSL e VAP foram maiores (P<0,05) com 15µM do que com 50 e100; WOB foi menor (P<0,05) com 100µM do que com 0, 15 e 25; e BCF foi maior (P<0,05) com 75 e 100µM do que com 0. No experimento 2, a motilidade progressiva foi maior (P<0,05) com 0 e 15µM do que com 50 e 75; LIN foi menor (P<0,05) com 75 e 100µM do que com 0 e 15; WOB foi maior (P<0,05) com 0 e 15µM; e iMP foi maior (P<0,05) com 100µM do que com 0. Assim, (+)-catequina ou EGCG em altas concentrações inibem, transitoriamente, a cinética de espermatozoides congelados caprinos, e 100µM de EGCG preserva a iMP.(AU)

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