Free-radical production after post-thaw incubation of ram spermatozoa is related to decreased in vivo fertility
| Autor: | L. Anel-López, María Rocío Fernández-Santos, José Julián Garde, Manuel Ramón, Ana J. Soler, Alejandro Maroto-Morales, A. Bisbal, Enrique del Olmo, Pilar Jiménez-Rabadán, Olga García-Álvarez |
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| Přispěvatelé: | Junta de Comunidades de Castilla-La Mancha, Ministerio de Ciencia e Innovación (España), CSIC - Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA) |
| Rok vydání: | 2015 |
| Předmět: |
Male
Pregnancy Rate Reproductive immunology medicine.medical_treatment frozen semen Artificial insemination Reproductive technology Semen analysis Biology law.invention Andrology Endocrinology fluorescein (CM-H2DCFDA) Pregnancy law In Situ Nick-End Labeling Genetics medicine Animals Flow citometry Molecular Biology Incubation Cryopreservation Sheep medicine.diagnostic_test flow cytometry Extender artificial insemination Flow Cytometry Spermatozoa Sperm Semen Analysis Fertility Reproductive Medicine Immunology Sperm Motility Frozen semen Female Animal Science and Zoology Reactive Oxygen Species Spermatogenesis Semen Preservation Developmental Biology Biotechnology |
| Zdroj: | RUIdeRA. Repositorio Institucional de la UCLM instname Digital.CSIC. Repositorio Institucional del CSIC |
| Popis: | The aim of the present study was to evaluate the effect of sperm reactive oxygen species (ROS) production and DNA changes on male fertility. For that purpose, six rams with significantly different pregnancy rates were used; these were classified as having high fertility, i.e. 59.4% average pregnancy rate, or low fertility, i.e. 23.1% average pregnancy rate. Sperm quality was assessed after a two-step process of sample thawing followed by an incubation of 2h, either in the freezing extender (37°C) or after dilution in synthetic oviductal fluid (SOF; 38°C, 5%CO2). Sperm viability (YO-PRO-1), ROS production (5-(and-6)-chloromethyl-2′,7′-dichlorodihydrofluorescein acetyl ester (CM-H2DCFDA)) and undamaged chromatin (sperm chromatin structure assay, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling, chromomycin A3) were evaluated by flow cytometry. Although no significant differences in sperm viability were observed, our results showed increased ROS production during incubation in the freezing extender as well as in SOF medium. Comparison between fertility groups showed significant differences in ROS production after 2h of incubation for the two treatments. Regarding DNA integrity, our results showed no significant differences either between treatments and incubation times or fertility groups. Linear regression analysis showed that ROS production determined by CM-H2DCFDA was a good indicator parameter for in vivo male fertility of SOF-incubated samples, yielding a fair correlation between both parameters (r≤-0.92). These results indicate that detection of ROS production by CM-H2DCFDA and flow cytometry after 2h of incubation in SOF could be a useful procedure for predicting fertility of ram spermatozoa. This research was supported by Education and science Council (JCCM, Spain) grant (PAI09-0006-3806). Enrique del Olmo is supported by a grant for training of technical personnel associated with PTA2008-0858-P project (Spanish ministry of Science and Innovation, MICINN). Manuel Ramón is supported by the DOC-INIA program. |
| Databáze: | OpenAIRE |
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