Resumo
Abstract Coenzyme Q-10 (CoQ-10) is a cofactor for mitochondrial electron transport chain and may be an alternative to improve sperm quality of cryopreserved equine semen. This work aimed to improve stallion semen quality after freezing by adding CoQ-10 to the cryopreservation protocol. Seven saddle stallions were utilized. Each animal was submitted to five semen collections and freezing procedures. For cryopreservation, each ejaculate was divided in three treatments: 1) Botucrio® diluent (control); 2) 50 μmol CoQ-10 added to Botucrio® diluent; 3) 1 mmol CoQ-10 added to Botucrio® diluent. Semen batches were analyzed for sperm motility characteristics (CASA), plasma and acrosomal membranes integrity and mitochondrial membrane potential (by fluorescence probes propidium iodide, Hoechst 33342, FITC-PSA and JC-1, respectively), alterations in cytoskeletal actin (phalloidin-FITC) and mitochondrial function (diaminobenzidine; DAB). The 1 mmol CoQ-10 treatment presented higher (P<0.05) amount (66.8%) of sperm cells with fully stained midpiece (indicating high mitochondrial activity) and higher (P<0.05) amount (81.6%) of cells without actin reorganization to the post-acrosomal region compared to control group (60.8% and 76.0%, respectively). It was concluded that the addition of 1 mmol CoQ-10 to the freezing diluent was more effective in preserving mitochondria functionality and cytoskeleton of sperm cells submitted to cryopreservation process.
Resumo
Coenzyme Q-10 (CoQ-10) is a cofactor for mitochondrial electron transport chain and may be an alternative to improve sperm quality of cryopreserved equine semen. This work aimed to improve stallion semen quality after freezing by adding CoQ-10 to the cryopreservation protocol. Seven saddle stallions were utilized. Each animal was submitted to five semen collections and freezing procedures. For cryopreservation, each ejaculate was divided in three treatments: 1) Botucrio® diluent (control); 2) 50 μmol CoQ-10 added to Botucrio® diluent; 3) 1 mmol CoQ-10 added to Botucrio® diluent. Semen batches were analyzed for sperm motility characteristics (CASA), plasma and acrosomal membranes integrity and mitochondrial membrane potential (by fluorescence probes propidium iodide, Hoechst 33342, FITC-PSA and JC-1, respectively), alterations in cytoskeletal actin (phalloidin-FITC) and mitochondrial function (diaminobenzidine; DAB). The 1 mmol CoQ-10 treatment presented higher (P<0.05) amount (66.8%) of sperm cells with fully stained midpiece (indicating high mitochondrial activity) and higher (P<0.05) amount (81.6%) of cells without actin reorganization to the post-acrosomal region compared to control group (60.8% and 76.0%, respectively). It was concluded that the addition of 1 mmol CoQ-10 to the freezing diluent was more effective in preserving mitochondria functionality and cytoskeleton of sperm cells submitted to cryopreservation process.(AU)
Assuntos
Animais , Masculino , Cavalos/genética , Ubiquinona/administração & dosagem , Dinâmica Mitocondrial , Criopreservação , Espermatozoides/química , ActinasResumo
The testicular artery is responsible for the blood supply that reaches the testis and has great importance in heat radiation. Vascular changes in the testis may lead to damage in sperm production, reflected in sperm motility and morphology. The aim of the present study was to evaluate correlations between testicular vascularity and sperm characteristic. Eight adult Santa Ines rams showing different reproductive status were used. The testicular vascularity and sperm characteristics were evaluated fortnightly during 90 days. Color Doppler ultrasonography was used to evaluate the testicular hemodynamic. Resistance index (RI) and pulsatility index (PI) of the testicular artery were evaluated by spectral-Doppler mode. The color-Doppler mode was used to evaluate the blood flow of the pampiniform plexus and testicular parenchyma. The semen analyses assessed were volume, concentration, motility, and morphology. The data were submitted to Pearson´s linear correlations test (p 0.05 was considered significant). No correlations were found between motility and testicular hemodynamic. The percentage of total sperm defects was positively correlated to left and right parenchymal score and to left RI and PI. On the other hand, the pampiniform plexus score was positively correlated to the number of colored pixels and negatively correlated to the RI and PI, for both sides. This study showed that the increase of sperm defect can be related to increase of testicular blood flow; however, more studies are need(AU)
A artéria testicular é responsável pelo fluxo de sangue que chega aos testículos e tem grande importância na termorregulação. Mudanças vasculares nos testículos podem levar à queda da produção espermática, refletindo na motilidade e morfologia. O objetivo deste trabalho foi avaliar as correlações entre a vascularização testicular e as características espermáticas. Foram utilizados oito carneiros adultos Santa Inês com diferentes status reprodutivos. A vascularização testicular e as características seminais foram avaliadas por um período de 90 dias. A ultrassonografia Doppler colorida foi utilizada para avaliar a hemodinâmica testicular. Foram calculados os índices de resistência (RI) e os índices de pulsatilidade (PI) com o modo espectral do Doppler. O modo colorido do Doppler foi utilizado para analisar o fluxo sanguíneo do plexo pampiniforme e do parênquima testicular. As características seminais avaliadas foram o volume, concentração, motilidade e morfologia. Os dados foram submetidos ao teste de correlação linear de Person (P 0.05 foi considerado significativo). Não foram encontradas relações entre motilidade e a hemodinâmica testicular. A porcentagem de defeitos totais correlacionou-se positivamente com o escore de vascularização dos parênquimas direito e esquerdo, e com o RI e PI esquerdos. Também o escore de vascularização dos plexos se correlacionou positivamente com a média de pixels e negativamente com o RI e PI, de ambos os lados. Este trabalho mostrou que o aumento de defeitos espermáticos pode estar correlacionado com o aumento do fluxo sanguíneo nos testículos; contudo, mais estudos são necessários(AU)