Vitrification induces critical subcellular damages in ram spermatozoa |
| |
| Affiliation: | 1. Department of Genetics, University of Cordoba, 14014, Cordoba, Spain;2. Instituto de Investigación y Formación Agraria y Pesquera (IFAPA) Hinojosa Del Duque, Carretera el Viso, Km 2, 14270, Cordoba, Spain;3. Centro Agropecuario Provincial de la Diputación de Cordoba, 14014, Cordoba, Spain;4. Department of Animal Medicine and Surgery, University of Cordoba, 14014, Cordoba, Spain;1. University of Namur, Faculty of Sciences, Department of Veterinary Medicine, Integrated Veterinary Research Unit, Namur Research Institute for Life Sciences (NARILIS), Rue de Bruxelles 61, B-5000, Namur, Belgium;2. Ecole Nationale d’Agriculture de Meknès, Department of Animal Production, Route Haj Kaddour, BP. S/40, 50001, Meknes, Morocco;3. Institut National de la Recherche Agronomique, Regional Center of Tangier, Bd Sidi Mohamed Ben abdellah78, 90010, Tangier, Morocco;1. Laboratorio de Teriogenología “Dr. Héctor H. Morello”, Instituto de Biotecnología Agropecuaria del Comahue (IBAC). Facultad de Ciencias Agrarias, Universidad Nacional del Comahue, Cinco Saltos, Río Negro, Argentina;2. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Cinco Saltos, Río Negro, Argentina;3. Ministerio de Desarrollo Territorial, Gobierno de la Provincia del Neuquén, Neuquén, Argentina;1. Departamento de Reproducción Animal, SGIT-INIA, Avda. Puerta de Hierro km 5.9, 28040, Madrid, Spain;2. Cnpq, Brazil;1. Department of Genetics, Faculty of Veterinary Medicine, University of Cordoba, Cordoba, 14014, Spain;2. Instituto de la Grasa, Consejo Superior de Investigaciones Científicas (CSIC), Sevilla, 41013, Spain;3. Centro Agropecuario Provincial de la Diputación de Cordoba, Cordoba, 14014, Spain;4. Department of Animal Medicine and Surgery, Faculty of Veterinary Medicine, University of Cordoba, Cordoba, 14014, Spain;1. Department of Animal Science, College of Agriculture, University of Tabriz, Tabriz, Iran;2. Department of Animal Science, University of Illinois, Urbana, IL, USA;1. Department of Technology, Postgraduate studies in Bioprocess Engineering and Biotechnology, Human and Animal Health, Federal University of Paraná, Rua dos Funcionários, 1540, Curitiba, Paraná, Brazil;2. Department of Veterinary Medicine, Postgraduate Studies in Animal Science, Pontifical Catholic University of Paraná, Rua Imaculada Conceição, 1155, Prado Velho, Curitiba, Paraná, Brazil;3. Department of Animal Reproduction and Veterinary Radiology-FMVZ − UNESP, Distrito de Rubiao Júnior, s/n, CEP: 18618-970, Botucatu, São Paulo, Brazil |
| |
| Abstract: | The aim of the present study was to analyse morphological variations in ovine spermatozoa subjected to different cryopreservation protocols using high resolution imaging techniques. Ejaculates were pooled and diluted in Tris-based extender. Aliquots containing 300 × 106 spz/ml were prepared and evaluated a) after the semen collection and pooling, b) after conventional freezing, c) after vitrification of samples maintained at room temperature (22 °C) prior to vitrification, and d) after vitrification of samples maintained at 5 °C prior to vitrification. Sperm motility, acrosome integrity, DNA fragmentation and morphology were assessed. Subcellular sperm changes were assessed and described by light microscopy, scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The maintenance of spermatozoa at 5 °C prior to vitrification and the use of 0.4 M sucrose pointed out lower dimensions of area, length and width than fresh, frozen and sperm maintained at 22 °C prior to vitrification. It was observed that the head width and length are significantly higher (P < 0.0001) in fresh spermatozoa than in the vitrified sperm samples. It could be hypothesized that greater intracellular fluid loss during vitrification could prevent damages in the spermatozoon throughout the reduced ice crystals formation, but mainly by the reduction of extracellular ice crystals due to the physical properties modification obtained when high concentrations of sugars are added. This is the first ultramicroscopic study carried out in ovine vitrified spermatozoa, which confirms the functional sperm alterations previously detected. |
| |
| Keywords: | Cryopreservation Ovine Spermatozoa Ultrastructure Vitrification |
| 本文献已被 ScienceDirect 等数据库收录！ |
|
