Treatment with chemical delipidation forskolin prior to cryopreservation improves the survival rates of swamp buffalo (Bubalus bubalis) and bovine (Bos indicus) in vitro produced embryos |
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| Affiliation: | 1. Department of Obstetrics, Gynaecology and Reproduction, Faculty of Veterinary Sciences, Chulalongkorn University, Bangkok, 10330, Thailand;2. Faculty of Veterinary Science, Prince of Songkla University, Songkhla, 90110, Thailand;3. Department of Animal Science, Faculty of Agriculture, Khon Kaen University, Khon Kaen, 40002, Thailand;1. NARO Institute of Livestock and Grassland Science, Tsukuba, Ibaraki 305-0901, Japan;2. Rakuno Gakuen University, Ebetsu, Hokkaido 069-8501, Japan;3. College of Agriculture and Life Sciences, Seoul National University, Seoul 151-742, Republic of Korea;4. Food and Fertilizer Technology Center, Taipei 10648, Taiwan;1. Department of Reproduction and Artificial Insemination, Faculty of Veterinary Medicine, Cumhuriyet University, Sivas, Turkey;2. Department of Reproduction and Artificial Insemination, Faculty of Veterinary Medicine, Ondokuz Mayıs University, Samsun, Turkey;1. Department of Animal Science, Division of Applied Life Science, Gyeongsang National University, Jinju, Gyeongnam Province 660-701, Republic of Korea;2. Institute of Agriculture and Life Science, Gyeongsang National University, Jinju, Gyeongnam Province 660-701, Republic of Korea;3. Animal, Dairy, and Veterinary Sciences Department, Utah State University, Logan, UT 84322-4700, USA;4. School of Veterinary Medicine, Utah State University, Logan, UT 84322-4700, USA;5. Center for Integrated Biosystems, Utah State University, Logan, UT 84322-4700, USA;1. School of Agriculture and Veterinary Medicine, University of Brasilia, Brasília, Federal District, Brazil;2. Centre de Recherche en Biologie de la Reproduction, Faculté des Sciences de l’Agriculture et de l’Alimentation, Département des Sciences Animales, Pavillon INAF, Université Laval, Québec City, Quebec, Canada;3. Embrapa Genetic Resources and Biotechnology, Laboratory of Animal Reproduction, Brasília, Federal District, Brazil;1. Laboratory of Physiology of Reproduction, Department of Animal Health, School of Veterinary Medicine, UNESP—Universidade Estadual Paulista, Araçatuba, São Paulo, Brazil;2. ThoMSon Mass Spectrometry Laboratory, Chemistry Institute, University of Campinas (UNICAMP), Campinas, São Paulo, Brazil;3. Brazilian Agricultural Research Corporation, EMBRAPA Pantanal, Corumbá, Mato Grosso do Sul, Brazil |
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| Abstract: | The cryopreservation of embryos is a technology developed for long-term genetic preservation. However, high sensitivity to low temperatures due to a large number of intracellular lipids within ruminant embryos compromises the success of this technique. The aim of this study was to examine the effects of using of lipolytic chemical agent forskolin, during in vitro producing of buffalo and bovine embryos on lipid contents, cryotolerance and subsequent developmental competence of these embryos. Buffalo and bovine oocytes were collected by the aspiration technique from follicles and submitted for in vitro fertilisation; the embryos were later divided into four experiments. Experiment 1, buffalo and bovine embryos were pre-treated in the presence and absence of 10 μM forskolin for 24 h. Lipid contents were determined by Nile red staining and confocal microscopy. We found that 10 μM forskolin was capable to reduce lipid contents within developing embryos in both of species (P < 0.01). Lipid contents within Day 2 embryos exhibited greater fluorescence intensity than did Day 7 embryos in both animal species. The purpose of Experiment 2 was to investigate the adverse effects of 10 μM forskolin on embryo development. In Experiments 3 and 4, Day 2 (4- to 8-cell stage) and Day 7 (blastocyst stage) embryos were pre-treated with 10 μM forskolin for 24 h and further cryopreserved with a controlled-rate freezing technique. The successful cryopreservation was determined by post-thawed embryonic development in vitro. The results showed that the blastocyst rate of the 4–8 cell stage in the forskolin-treated group had increased in both species, while the hatching and hatched blastocyst rates of forskolin-treated day 7 bovine embryos were significantly higher than those of the non-treated group (52.1% vs. 39.4%; P < 0.05). However, delipidation with forskolin did not affect the developmental rate of the day 7 buffalo embryos (P = 0.73). Our studies showed that delipidation by forskolin treatment increased the survival rate of cryopreservation in buffalo and bovine in vitro produced embryos. |
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| Keywords: | Delipidation Forskolin Cryotolerance IVP embryos Bovine Swamp buffalo |
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