ABSTRACT
The present study analyzed the changes in gene expression induced by the Cryotop vitrification technique in bovine blastocyst-stage embryos, using Agilent EmbryoGENE microarray slides. Bovine in vitro-produced embryos were vitrified and compared with nonvitrified (control) embryos. After vitrification, embryos were warmed and cultured for an additional 4 hours. Survived embryos were used for microarray analysis and quantitative polymerase chain reaction (qPCR) quantification. Survival rates were higher (P < 0.05) in the control embryos (100%) than in the vitrified embryos (87%). Global gene expression analysis revealed that only 43 out of 21,139 genes exhibited significantly altered expression in the vitrified embryos compared to the control embryos, with a very limited fold change (P < 0.05). A total of 10 genes were assessed by qPCR. Only the FOS-like antigen 1 (FOSL1) gene presented differential expression (P < 0.05) according to both the array and qPCR methods, and it was overexpressed in vitrified embryos. Although, the major consequence of vitrification seems to be the activation of the apoptosis pathway in some cells. Indeed, FOSL1 is part of the activating protein 1 transcription factor complex and is implicated in a variety of cellular processes, including proliferation, differentiation, and apoptosis. Therefore, our results suggest that a limited increase in the rate of apoptosis was the only detectable response of the embryos to vitrification stress.
Subject(s)
Cattle/embryology , Cryopreservation/veterinary , Fertilization in Vitro/veterinary , Gene Expression Regulation, Developmental/physiology , Transcriptome , Vitrification , Animals , Blastocyst/physiology , Embryo Culture Techniques , Polymerase Chain Reaction/methods , Polymerase Chain Reaction/veterinary , Protein Array Analysis/veterinaryABSTRACT
Para avaliar a digestibilidade in vitro da jitirana com inóculo cecal de avestruzes, usou-se delineamento inteiramente casualizado em esquema fatorial (4x2), com quatro diferentes idades de corte (60; 75; 90 e 105 dias) e duas formas de uso, in natura ou em feno. Houve interação significativa para digestibilidade in vitro da matéria seca e de frações proteicas da jitirana, com média total de 59,57% para matéria seca; 28,07% proteína bruta e 21,70% para proteína insolúvel em detergente neutro. Não foi observada interação significativa para a digestibilidade in vitro da fração fibrosa da jitirana, com média total de 45,20% para fibra em detergente neutro e 49,83% fibra em detergente neutro corrigida para cinzas e proteína. Em relação às diferentes idades de corte da jitirana, para a planta in natura, as médias de digestibilidade in vitro da matéria seca variaram entre 71,75 e 53,87%, enquanto, para a planta fenada, as médias variaram entre 61,36 e 50,23%. Tanto para a planta in natura quanto para o feno de jititrana, a idade de corte aos 60 dias propiciou melhor digestibilidade, especialmente em comparação aos 105 dias, em virtude de uma melhor qualidade e disponibilidade nutricional para atividade fermentativa cecal.
To evaluated the in vitro digestibility of jitirana using ostriches ceacal liquor, it were used entirely randomized design on factorial schedule (4x2), with four cut age of forage (60; 75; 90 and 105 days old) and two types forage in natura or hay. There was a significative interaction between cut age and jitirana type to in vitro digestibility of dry matter and proteic fractions, and the means obtained were 59.57% to dry matter, 28.07% to crude protein and 21.7% to neutral detergent insoluble protein. It were not observed significative interaction to in vitro digestibility of fiber fraction, and the means obtained were 45.20% to neutral detergent fiber and 49.83% to neutral detergent fiber corrected to ash and protein. In relation to the different cut age of jitirana plant, to the in vitro digestibility of dry matter to the in natura type, the means stayed between 71.75 and 53.87%, while to the jitirana hay type the in vitro digestibility of dry matter the means stayed between 61.36 and 50.23%. For both jitirana type in natura or hay, the cut age on 60 days old propitiated the best values of digestibility when compared to the cut age on 105 days old, as a higher available and nutritional quality to the fermentative caecal activity.
Subject(s)
Animals , Protein Array Analysis/veterinary , Plant Structures/adverse effectsABSTRACT
Para avaliar a digestibilidade in vitro da jitirana com inóculo cecal de avestruzes, usou-se delineamento inteiramente casualizado em esquema fatorial (4x2), com quatro diferentes idades de corte (60; 75; 90 e 105 dias) e duas formas de uso, in natura ou em feno. Houve interação significativa para digestibilidade in vitro da matéria seca e de frações proteicas da jitirana, com média total de 59,57% para matéria seca; 28,07% proteína bruta e 21,70% para proteína insolúvel em detergente neutro. Não foi observada interação significativa para a digestibilidade in vitro da fração fibrosa da jitirana, com média total de 45,20% para fibra em detergente neutro e 49,83% fibra em detergente neutro corrigida para cinzas e proteína. Em relação às diferentes idades de corte da jitirana, para a planta in natura, as médias de digestibilidade in vitro da matéria seca variaram entre 71,75 e 53,87%, enquanto, para a planta fenada, as médias variaram entre 61,36 e 50,23%. Tanto para a planta in natura quanto para o feno de jititrana, a idade de corte aos 60 dias propiciou melhor digestibilidade, especialmente em comparação aos 105 dias, em virtude de uma melhor qualidade e disponibilidade nutricional para atividade fermentativa cecal. (AU)
To evaluated the in vitro digestibility of jitirana using ostriches ceacal liquor, it were used entirely randomized design on factorial schedule (4x2), with four cut age of forage (60; 75; 90 and 105 days old) and two types forage in natura or hay. There was a significative interaction between cut age and jitirana type to in vitro digestibility of dry matter and proteic fractions, and the means obtained were 59.57% to dry matter, 28.07% to crude protein and 21.7% to neutral detergent insoluble protein. It were not observed significative interaction to in vitro digestibility of fiber fraction, and the means obtained were 45.20% to neutral detergent fiber and 49.83% to neutral detergent fiber corrected to ash and protein. In relation to the different cut age of jitirana plant, to the in vitro digestibility of dry matter to the in natura type, the means stayed between 71.75 and 53.87%, while to the jitirana hay type the in vitro digestibility of dry matter the means stayed between 61.36 and 50.23%. For both jitirana type in natura or hay, the cut age on 60 days old propitiated the best values of digestibility when compared to the cut age on 105 days old, as a higher available and nutritional quality to the fermentative caecal activity. (AU)