Effect of single and combined treatments with MPF or MAPK inhibitors on parthenogenetic haploid activation of bovine oocytes |
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| Institution: | 1. Universidad de Alicante, Departamento de Biotecnología, Alicante, Spain;2. FISABIO - Hospital Universitario de San Juan de Alicante, Servicio de Ginecología y Obstetricia, San Juan de Alicante, Spain;3. Hospital Universitario San Juan de Alicante, Servicio de Ginecología y Obstetricia, San Juan de Alicante, Spain;4. Universidad Miguel Hernández de Elche, Medicina clínica, San Juan de Alicante, Spain;5. Universidad Miguel Hernández de Elche, División de Ginecología, San Juan de Alicante, Spain;6. Cátedra Human Fertility, Universidad de Alicante, Alicante, Spain;1. Institute of Biology, University of Brasília, Brasília, Brazil;2. Embrapa Genetic Resources and Biotechnology, Brasília, Brazil;3. Veterinary Medicine Department, University of Espírito Santo, Alegre, Brazil;1. Department of Animal Science, National Chung Hsing University, Taichung 402, Taiwan;2. Graduate Institute of Basic Medical Science, China Medical University, Taichung 402, Taiwan;3. Graduate Institute of Veterinary Pathobiology, College of Veterinary Medicine, National Chung Hsing University, Taichung 402, Taiwan;4. Department of Animal Science and Biotechnology, Tunghai University, Taichung 407, Taiwan;5. Faculty of Veterinary Medicine, University of Calgary, Calgary, AB, Canada;6. Agriculture Biotechnology Center, National Chung Hsing University, Taichung 402, Taiwan;7. Medical Research Department, China Medical University Hospital, Taichung 404, Taiwan;8. Department of Biomedical Informatics, College of Computer Science, Asia University, Taichung 413, Taiwan;1. State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangxi University, Nanning, Guangxi, 530004, China;2. Laboratory Animal Center, Guangxi Medical University, Nanning, Guangxi, 530021, China |
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| Abstract: | In bovine, correct oocyte artificial activation is a key step in ICSI and other reproductive biotechnologies, and still needs to be improved. The current study was designed to compare the activating efficiency of ionomycin (Io) followed by: a 4 h time window and ethanol (4h-Et), roscovitine (Rosc), dehydroleucodine (DhL), cycloheximide (CHX) or PD0325901 (PD), each as a single treatment, and then combine them in novel protocols. Parthenogenetic haploid activation was evaluated in terms of pronuclear (PN) formation, second polar body (2PB) extrusion, ploidy of day 2 embryos and in vitro development. Combined treatments with Io-4h-Et-Rosc and Io-Rosc/CHX increased PN formation (92.2% and 96%, respectively) compared with Io-Rosc, Io-CHX or Io-4h-Et, which were equally efficient at inducing PN formation (82–84%) and 2PB extrusion (62.1–70.5%). Oocyte activation with Io-DhL and Io-Rosc/DhL resulted in higher 2PB extrusion rates (90% and 95.9%, respectively) but lower PN formation (49.4–58.8%) and cleavage rates (36–57.9%), as occurred with Io-CHX/DhL (76.4% and 70.4%, respectively). For the first time, results show that Io followed by the MAPK inhibitor PD induces PN formation and 2PB extrusion, but PD combined with Rosc or CHX resulted in low rates of haploid day 2 embryos. In conclusion, DhL strongly induces 2PB extrusion but leads to poor PN formation and embryo development. PD induces bovine oocyte activation but results in low rates of haploid embryos. In contrast, the improved PN formation rates after treatment with combined Io-4h-Et-Rosc and Io-Rosc/CHX suggest they should be further evaluated in ART, aiming to increase success rates in bovine. |
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| Keywords: | Parthenogenesis ART Cattle Meiosis |
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