Expression profile of key genes involved in DNA repair mechanisms in bovine cumulus cells cultured with bovine serum albumin or fetal calf serum |
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| Affiliation: | 1. Centro de Ciências Rurais, Universidade Federal de Santa Catarina (UFSC), Curitibanos, SC, Brazil;2. Department of Animal Sciences, University of Florida, Gainesville, FL, USA;3. Laboratório de Reprodução Animal Assis Roberto de Bem, Centro de Ciências Agroveterinárias (CAV), Universidade do Estado de Santa Catarina (UDESC), Lages, SC, Brazil;4. Centre de Recherche en Reproduction Animale, Faculté de Médecine Vétérinaire, Université de Montréal, St-Hyacinthe, Quebec, Canada;5. Laboratório de Biotecnologia e Reprodução Animal (BIOREP), Universidade Federal de Santa Maria (UFSM), Santa Maria, RS, Brazil;1. Division of Animal Science, ICAR-Central Island Agricultural Research Institute, Port Blair 744105, Andaman and Nicobar Islands, India;2. Animal Physiology and Reproduction, ICAR-National Research Centre on Mithun, Medziphema 797106, Nagaland, India;3. Division of Animal Physiology, ICAR-National Dairy Research Institute, Karnal 13200, Haryana, India;1. Department of Theriogenology, Faculty of Veterinary Medicine, Assiut University, Assiut, Egypt;2. Department of Theriogenology, Faculty of Veterinary Medicine, Benha University, Egypt;3. Department of Animal Reproduction and A.I, Veterinary Research Division, National Research Centre, Dokki, 12622 Giza, Egypt;4. Joint Faculty of Veterinary Medicine, Yamaguchi University, Yamaguchi 1677-1, Japan;5. Global Agromedicine Research Center (GAMRC), Obihiro University of Agriculture and Veterinary Medicine, Obihiro 080 8555, Japan;1. Departamento de Fisiología y Desarrollo Celular, Instituto Nacional de Perinatología “Isidro Espinosa de los Reyes”, México;2. Coordinación de Salud Mental, Instituto Nacional de Perinatología “Isidro Espinosa de los Reyes”, México;3. Unidad de Investigación en Reproducción Humana, Instituto Nacional de Perinatología-Facultad de Química, Universidad Nacional Autónoma de México, México;1. Key Laboratory of Adaptation and Evolution of Plateau Biota, Northwest Institute of Plateau Biology, Chinese Academy of Sciences, Xining 810001, China;2. University of Chinese Academy of Sciences, Beijing 100049, China;3. Qinghai Key Laboratory of Animal Ecological Genomics, Northwest Institute of Plateau Biology, Chinese Academy of Sciences, Xining 810001, China;1. Savaid Stomatology School, Hangzhou Medical College, Hangzhou 310053, China;2. Bourn Hall Reproductive Medical Center, Kunming City Maternal and Child Health Hospital, Kunming 650031, China;1. Laboratory of Animal Biotechnology, Department of Animal Science, Faculty of Veterinary Sciences, Universidad de Concepción, Chillán 3812120, Chile;2. Centro de investigación en Tecnología de Embriones, Facultad de Zootecnia, Universidad Nacional Agraria La Molina, La Molina, Lima, Peru;3. ANID-Millennium Science Initiative Program Millennium Nucleus of Ion Channels-Associated Diseases (MiNICAD), Center for Bioinformatics, Simulation and Modeling, CBSM, Departament of Bioinformatics, Faculty of Engineering, Campus Talca, University of Talca, Talca 3460000, Chile |
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| Abstract: | Cumulus cells from cumulus-oocyte complexes (COC) matured in vitro in serum-free medium show high incidence of apoptosis and DNA double-strand breaks (DSB). This study aimed to characterize the transcript expression profile of selected genes involved in DNA repair mechanisms in bovine cumulus cells cultured with bovine serum albumin (BSA) or fetal calf serum (FCS). Briefly, bovine cumulus-oocyte complexes were in vitro matured with either, 0.4% BSA or 10% FCS for 3, 6, 12 or 24 h. The total RNA of cumulus cells was used for real-time PCR analysis. Transcript abundance of XRCC6, XRCC5, DNAPK, GAAD45B, TP53BP1, RAD50, RAD52, ATM and BRCA2 target genes changed as the IVM proceeded (P < 0.05). However, an interaction between protein source (FCS or BSA) and time was not detected (P ≥ 0.05). Cumulus cells from COCs matured with BSA presented higher mRNA expression of two genes compared to FCS group: TP53BP1 at 6 h and BRCA1 at 3, 6, 12 and 24 h (P < 0.05). In summary, our results showed for the first time the expression profile of the key genes involved in DSB repair mechanisms in cumulus cells obtained from bovine COCs matured with FCS or BSA. The higher mRNA expression of BRCA1 and TP53BP1 and lower mRNA expression of TNFAIP6 suggests an increase in apoptosis rate and DNA damage in cumulus cells cultured in BSA-supplemented medium and may explain, at least to some extent, the reduced developmental potential of bovine oocytes matured in serum-free medium. |
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| Keywords: | In vitro maturation Cumulus cells DNA repair Apoptosis |
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