Analysis of selected sperm by density gradient centrifugation might aid in the estimation of in vivo fertility of thawed ram spermatozoa
| Autor: | Alejandro Maroto-Morales, Manuel Ramón, Ana J. Soler, Pilar Jiménez-Rabadán, A. E. Domínguez-Rebolledo, A. Bisbal, M.D. Pérez-Guzmán, Vidal Montoro, Olga García-Álvarez, E. del Olmo |
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| Přispěvatelé: | CSIC - Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Junta de Comunidades de Castilla-La Mancha, Ministerio de Educación y Ciencia (España) |
| Rok vydání: | 2010 |
| Předmět: |
Male
endocrine system Pregnancy Rate medicine.medical_treatment Statistics as Topic Semen Cell Separation Fertilization in Vitro Semen analysis Biology Insemination Male infertility Andrology Food Animals Pregnancy Density-gradient centrifugation Centrifugation Density Gradient medicine Animals Cryopreserved semen Spermatozoa selection Small Animals Infertility Male reproductive and urinary physiology Sperm motility Cryopreservation Sheep medicine.diagnostic_test urogenital system Equine Artificial insemination Sperm washing Flow Cytometry Prognosis medicine.disease Spermatozoa Sperm Semen Analysis Fertility Female Animal Science and Zoology Semen Preservation |
| Zdroj: | Digital.CSIC. Repositorio Institucional del CSIC instname |
| ISSN: | 0093-691X |
| DOI: | 10.1016/j.theriogenology.2010.04.027 |
| Popis: | The aim of the present study was to evaluate the effect of selecting a sperm subpopulation by means of a discontinuous density gradient centrifugation (DGC) on the quality of ram thawed semen, and the relationships between sperm parameters assessed in unselected and in selected sperm samples with in vivo fertility after intrauterine artificial insemination (IUI) using unselected sperm samples. Semen samples from twenty males were collected by artificial vagina and cryopreserved following a standard protocol. After thawing, unselected sperm samples were used in an in vivo fertility trial and sperm motility (subjective and objective, assessed by means of CASA) and membrane and acrosomal integrities (microscopy) were evaluated on unselected and selected sperm samples. In addition, plasmalemma integrity (YO-PRO-1/PI), membrane fluidity (Merocyanine 540/YO-PRO-1), mitochondrial activity (Mitotracker Deep Red/YO-PRO-1), and DNA fragmentation index (%DFI) assessed by Sperm Chromatin Structure Assay (SCSA®) were evaluated by flow cytometry before and after sperm processing using DGC. Results showed that DGC improved all sperm parameters significantly, except the %DFI, which increased after the selection procedure. No relationships were found between sperm parameters evaluated in unselected sperm samples and in vivo fertility. However, we found a positive correlation between spermatozoa with high membrane fluidity within the viable sperm population (VIABMerocyanine+) evaluated in selected sperm samples and in vivo fertility (r = 0.370, P = 0.019). In conclusion, our results suggest that selected spermatozoa represent a sperm subpopulation different to the unselected one that could be related with the in vivo fertility. This work was supported by Spanish Ministry of Education and Science (RTA2005-00091-CO2) and by Education and Science Council of Junta de Comunidades de Castilla-La Mancha (PCC08-0105). Olga García-Álvarez and Pilar Jiménez-Rabadán were recipients of scholarships from INIA and Alejandro Maroto-Morales from JCCM. |
| Databáze: | OpenAIRE |
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