Addition of cholesterol loaded cyclodextrin prior to GV-phase vitrification improves the quality of mature porcine oocytes in vitro |
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| Institution: | 1. Audubon Nature Center for Research of Endangered Species, New Orleans, LA, USA;2. Department of Biological Sciences, University of New Orleans, New Orleans, LA, USA;1. IVI Valencia, IVI-RMA Global, Plaza Policía Local, 3, 46015, Valencia, Spain;2. Embryotools, Parc Científic de Barcelona, Av. Doctor Marañon, 8, 08028, Barcelona, Spain;1. Department of Andrology, People’s Hospital of Liaocheng City, China;2. Department of Obstetrics and Gynecology, People’s Hospital of Linyi City, China;3. Department of Reproductive Medicine, Jinan Central Hospital Affiliated to Shandong University, China;4. Department of Reproductive Medicine, Hospital for Maternity and Child Care of Jinan City, China;1. Departamento de Ciencias de la Salud, División de Ciencias Biológicas y de la Salud, Universidad Autónoma Metropolitana-Iztapalapa, Av. San Rafael Atlixco 186, 09340 DF, México;2. Departamento de Biología de la Reproducción, División de Ciencias Biológicas y de la Salud, Universidad Autónoma Metropolitana-Iztapalapa, Av. San Rafael Atlixco 186, 09340 DF, México;3. Maestría en Biología de la Reproducción Animal, División de Ciencias Biológicas y de la Salud, Universidad Autónoma Metropolitana-Iztapalapa Av. San Rafael Atlixco 186, 09340 DF, México;1. Yunnan Animal Science and Veterinary Institute, Jindian, Panlong County, Kunming City, Yunnan Province, China;2. Yunnan Provincial Meat Caprine Engineering Research Center, Jindian, Panlong County, Kunming City, Yunnan Province, China;3. Yunnan Provincial Engineering Laboratory of Animal Genetic Resource Conservation and Germplasm Enhancement, Jindian, Panlong County, Kunming City, Yunnan Province, China;4. College of Animal Science and Technology, China Agricultural University, Yuanmingyuan West Road, Haidian County, Beijing, China |
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| Abstract: | The purpose of this study was to determine whether the mitochondrial membrane potential, pro-apoptotic gene expression, and ubiquitylation status of zona pellucida proteins (ZP1, ZP2, and ZP3) of vitrified GV-stage mature oocytes could be protected by treatment with cholesterol-loaded methyl-β-cyclodextrin (CLC) prior to vitrification. Porcine GV oocytes were treated with CLC prior to the vitrification process, and the effects on the mitochondrial membrane potential and ZP ubiquitylation status were determined by JC-1 single staining and western blot assays. We found that porcine GV-stage oocytes were treated with CLC at different concentrations (0.5, 5, and 10 mg/mL) prior to vitrification improved in vitro maturation of these oocytes (P < 0.05). The mitochondrial membrane potential of matured oocyte without vitrification or treated with 5 mg/mL CLC vitrification treatment was higher than that of the 0 mg/mL CLC group and other treatment groups (vitrified) (P < 0.05). The expression of Caspase 3, Caspase 8, and Caspase 9 genes in the high concentration CLC treatment groups (5 and 10 mg/mL) was significantly lower than that in the 0 (vitrified) mg/mL CLC group (P < 0.05). ZPs protein and ZP3 protein ubiquitylation were also higher in the non-vitrified controls, 5 and 10 mg/mL CLC-treated oocytes than in the 0 (vitrified) and 0.5 mg/mL vitrified groups (P < 0.05). Whereas the sperm–oocyte binding capacity was improved in the CLC treatment groups (P < 0.05) but the embryonic development rate was not improved. In conclusion, pretreatment with CLC can improve the survival rate and maturation rate of oocytes and protect their mitochondria and zona pellucida of porcine oocytes from cryodamage during the vitrification process. |
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| Keywords: | Porcine oocyte Vitrification CLC Mitochondria ZP ubiquitylation |
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