J J Dai1, Y F Niu1, C F Wu1, S H Zhang1, D F Zhang2. 1. Institute of Animal Science and Veterinary Science, Shanghai Academy of Agricultural Sciences; Division of Animal Genetic Engineering, Shanghai Municipal Key Laboratory of Agri-Genetics and Breeding, Shanghai, China. 2. Institute of Animal Science and Veterinary Science, Shanghai Academy of Agricultural Sciences; Division of Animal Genetic Engineering, Shanghai Municipal Key Laboratory of Agri-Genetics and Breeding, Shanghai, China. zhangdefuzdf@163.com.
Abstract
BACKGROUND: Oocytes vitrification is widely used for cryopreservation of female genetic resources. OBJECTIVE: In order to illuminate the apoptotic pathways of porcine MII stage oocytes after vitrification. MATERIALS AND METHODS: This study used in situ fluorescence staining and RT-PCR to detect the expression levels of some key molecules from death receptor and mitochondria mediated apoptotic pathways. RESULTS: (1) Early stage apoptosis were detected in both PI staining survival oocytes and PI staining dead oocytes. (2) The fluorescence intensity of caspase 8, caspase 9, caspase 3 and pan caspase from vitrified oocytes were 32.03, 16.56, 16.70 and 8.43 respectively, which were much higher than those from fresh oocytes (4.02, 4.83, 4.23 and 3.08, P < 0.05). (3) Not only the genes from death receptor mediated apoptotic pathway, but also from mitochondrial mediated apoptotic pathway were changed greatly. CONCLUSION: The death of porcine vitrified oocytes could be induced by apoptosis, both death receptor and mitochondria mediated apoptotic pathways participated the occurrence of apoptosis in porcine vitrified MII stage oocytes.
BACKGROUND: Oocytes vitrification is widely used for cryopreservation of female genetic resources. OBJECTIVE: In order to illuminate the apoptotic pathways of porcine MII stage oocytes after vitrification. MATERIALS AND METHODS: This study used in situ fluorescence staining and RT-PCR to detect the expression levels of some key molecules from death receptor and mitochondria mediated apoptotic pathways. RESULTS: (1) Early stage apoptosis were detected in both PI staining survival oocytes and PI staining dead oocytes. (2) The fluorescence intensity of caspase 8, caspase 9, caspase 3 and pan caspase from vitrified oocytes were 32.03, 16.56, 16.70 and 8.43 respectively, which were much higher than those from fresh oocytes (4.02, 4.83, 4.23 and 3.08, P < 0.05). (3) Not only the genes from death receptor mediated apoptotic pathway, but also from mitochondrial mediated apoptotic pathway were changed greatly. CONCLUSION: The death of porcine vitrified oocytes could be induced by apoptosis, both death receptor and mitochondria mediated apoptotic pathways participated the occurrence of apoptosis in porcine vitrified MII stage oocytes.