Stability of the cytoskeleton of matured buffalo oocytes pretreated with cytochalasin B prior to vitrification |
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| Institution: | 1. Department of Animal Science, College of Agriculture and Natural Resources, University of Tehran, Karaj, Iran;2. Department of Clinical Sciences, Faculty of Veterinary Medicine, Shahid Chamran University, Ahvaz, Iran;3. Department of Poultry Science, Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran;1. IVF Spain Alicante, Av. de Ansaldo, 13, 03540, Alicante, Spain;2. iGLS, Alicante, Spain, C/ Britania, 7, 03540, Alacante, Spain;3. Biotechnology Department, University of Alicante.C/San Vicente del Raspeig, 03690, San Vicente del Raspeig, Alicante, Spain;1. University of Seville, C/ Camino de los Descubrimientos s/n, 41092, Seville, Spain;2. Ginemed Clínicas, C/ Farmacéutico Murillo Herrera nº 3-5, 41010, Seville, Spain;3. National Accelerator Centre, C/ Thomas Alva Edison 7, 41092, Seville, Spain;1. Department of Biology, University of Southern Denmark, Campusvej 55, 5230 Odense M, Denmark;2. Department of Science and Environment, Roskilde University, Universitetsvej 1, 4000 Roskilde, Denmark;1. Laboratory of Wild Animal Biology and Medicine, Federal University of Pará, Castanhal, PA, CEP 68740-970, Brazil;2. National Primate Center, Ananindeua, PA, CEP 66645-000, Brazil |
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| Abstract: | Stabilizing the cytoskeleton system during vitrification can improve the post-thaw survival and development of vitrified oocytes. The cytoskeleton stabilizer cytochalasin B (CB) has been used in cryopreservation to improve the developmental competence of vitrified oocytes. To assess the effect of pretreating matured buffalo oocytes with CB before vitrification, we applied 0, 4, 8, or 12 μg/mL CB for 30 min. The optimum concentration of CB treatment (8 μg/mL for 30 min) was then used to evaluate the distribution of microtubules and microfilaments, the expression of the cytoskeleton proteins actin and tubulin, and the developmental potential of matured oocytes that were vitrified-warmed by the Cryotop method. Western blotting demonstrated that vitrification significantly decreased tubulin expression, but that the decrease was attenuated for oocytes pretreated with 8 μg/mL CB before vitrification. After warming and intracytoplasmic sperm injection, oocytes that were pretreated with 8 μg/mL CB before vitrification yielded significantly higher 8-cell and blastocyst rates than those that were vitrified without CB pretreatment. The values for the vitrified groups in all experiments were significantly lower (P < 0.01) than those of the control groups. In conclusion, pretreatment with 8 μg/mL CB for 30 min significantly improves the cytoskeletal structure, expression of tubulin, and development capacity of vitrified matured buffalo oocytes. |
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| Keywords: | Vitrification Buffalo oocytes Cytochalasin B Cytoskeleton |
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