Effect of different glycerol concentrations on phosphatidylserine translocation and mitochondrial membrane potential in chilled boar spermatozoa |
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| Institution: | 1. Departamento de Producción Agropecuaria, Facultad de Ciencias Agropecuarias y Forestales, Universidad de La Frontera, Temuco, Chile;2. Centro de Biotecnología de la Reproducción – Núcleo Científico y Tecnológico en Biorecursos (CEBIOR-BIOREN), Universidad de La Frontera, Temuco, Chile;3. Departamento de Reprodução Animal, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, Pirassununga, São Paulo, Brazil;4. Departamento de Ingeniería Química, Facultad de Ingeniería y Ciencias, Universidad de La Frontera, Temuco, Chile;1. Department of Animal Reproduction, Gynecology and Obstetrics, College of Veterinary Science, AAU, Assam, India;2. National Research Centre on Pig (ICAR), Assam, India;3. Department of Veterinary Physiology, College of Veterinary Science, AAU, Assam, India;4. Department of Animal Genetics and Breeding, College of Veterinary Science, AAU, Assam, India;1. Corelab, Laboratorio Unico di AVR, Cesena (FC), Italy;2. Department of Anesthesia, G. Fracastoro Hospital, San Bonifacio (VR), Italy;3. Department of Pathology, Infectious Diseases Unit, Azienda Ospedaliera Universitaria Integrata, Verona, Italy;1. State Key Laboratory of Reproductive Regulation and Breeding of Grassland Livestock (R2BGL), Inner Mongolia University, 24 Zhaojun Rd., Hohhot, 010070, China;2. College of Life Sciences, Inner Mongolia University, 24 Zhaojun Rd., Hohhot, 010070, China;3. Xilingol Vocational College, No.11 Mingantu Street, Xilinhot, 026000, China |
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| Abstract: | Boar spermatozoa are extremely sensitive to low temperatures and the cryopreservation causes dramatic changes in sperm survivability, but it is not clear which part of the cryopreservation process affects the most. The aim of this work was to assess early events of apoptotic changes as damage indicators in boar sperm cooled to 5 °C and exposed to different glycerol (GLY) concentrations. For this purpose, progressive sperm motility (CASA), plasmatic and acrosome membranes integrity (CFDA/PI; phase contrast), plasma membrane functionality (HOS), phosphatidylserine translocation (Annexin-V/FITC) and reduction of mitochondrial membrane potential (Ψm) (JC-10) were carried out at 37 °C, 17 °C and 5 °C in eight boar sperm pools. Afterwards, three aliquots were diluted in different freezing extenders (control: 0% GLY; A: 2% GLY and B: 3% GLY); sperm quality and early apoptotic changes were assessed. Motility was negatively affected during cooling to 5 °C. Furthermore, plasma membrane functionality was the most affected by cooling. The number of necrotic cells was higher at 5 °C. However, no differences were observed in phosphatidylserine translocation. The extender with 3% GLY at 5 °C presented better Ψm than 0 and 2% GLY. Based on this analysis, boar sperm cooling to 5 °C does not modify the rate of early apoptotic changes, although alterations in the ?m were evident. |
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| Keywords: | Boar spermatozoa Refrigeration Apoptotic process Glycerol |
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