El papel del glutatión reducido (GSH) en la capacitación espermática y su relación con la fertilización in vitro en espermatozoides de cerdo sus Scrofa Domesticus
In TRA that seek to preserve sperm quality, GSH has been used as an antioxidant, trying to extend the lifespan of the male gamete. Although ROS cause cell damage, it is also clear that they play an important physiological role in sperm during SC which in turn affects fertilization. Therefore it is n...
| Autor: | |
|---|---|
| Tipo de recurso: | tesis de maestría |
| Estado: | Versión publicada |
| Fecha de publicación: | 2019 |
| País: | México |
| Institución: | Universidad Autónoma Metropolitana |
| Repositorio: | Repositorio Institucional de la UAM Iztapalapa |
| Idioma: | español |
| OAI Identifier: | oai:bindani.izt.uam.mx:x059c736d |
| Acceso en línea: | https://doi.org/10.24275/uami.x059c736d |
| Access Level: | acceso abierto |
| Palabra clave: | info:eu-repo/classification/LEM/Animals -- Reproduction info:eu-repo/classification/LEM/Fertilization in vitro info:eu-repo/classification/LEM/Animales -- Reproducción info:eu-repo/classification/LEM/Fertilización in vitro info:eu-repo/classification/LEM/Swine info:eu-repo/classification/LEM/Glutathione info:eu-repo/classification/LEM/Glutation info:eu-repo/classification/LEM/Cerdos info:eu-repo/classification/cti/2 |
| Sumario: | In TRA that seek to preserve sperm quality, GSH has been used as an antioxidant, trying to extend the lifespan of the male gamete. Although ROS cause cell damage, it is also clear that they play an important physiological role in sperm during SC which in turn affects fertilization. Therefore it is necessary to maintain a certain balance of both antioxidants and oxidants so that the SC is carried out correctly. The purpose of this study was to add determining if different concentrations of GSH during sperm capacitation in vitro fertilization porcine oocyte is modified. During SC, the relevant role of GSH has been demonstrated, reporting that at concentrations of 1 mM an adequate percentage of trained sperm is obtained, while at other concentrations, the training process is affected; this allows us to suppose that the GSH is an important SC regulator, For what is important to analyze, if the concentration of GSH administered during training, can modify IVF. For the methodology, a seminal analysis of the semen sample was first performed, evaluating viability, morphology concentration and mobility. subsequently the SC was performed for it took 5 mL of sample and was diluted and washed in DPBS, then it was placed in the middle apacitante c and incubated for 2 and 4 h with different concentrations of GSH to be evaluated (0, 0.5, 1 and 5 mM). For oocyte maturation, and punctured s ovaries of prepubertal bristle collecting oocytes, these were placed in maturation medium and incubated for 44h. Finally for fertilization , the matured oocytes were stripped for insemination with previously trained spermatozoa and determining different parameters such as: SC time and coincubation, fertilization medium: TBMm and TALP-HEPES, evaluate the IVF with the different concentrations of GSH (0, 0.5, 1 and 5 mM) and finally the coincubation of both gametes in GSH washed sperm. For the evaluation of SC time and coincubation it was found that at 2h of SC plus 4h of coincubation a percentage between 50 to 60% of IVF is obtained, unlike 4 h of SC and 2h of coincubation where the percentage was 0%. In the media evaluation, it was found that the TBMm had 53% IVF whereas in TALP-HEPES, 47% IVF was obtained without determining differences. For the washing of the sample, 53% of IVF were obtained in sperm without an additional wash, while those that were washed, the percentage of IVF was 0%. For the evaluation of IVF with the different concentrations of GSH, it was obtained in the concentration of 0.5, 1 and 5 mM a percentage of 42, 56 and 0% respectively of IVF obtaining higher percentages in the concentration of 1 mM. And finally for the coincubation of gametes with the different concentrations of GSH, a percentage of 54, 39 and 4% respectively of IVF was obtained in the concentration of 0.5, 1 and 5 mM, obtaining higher percentages in 0.5 without the concentration of 1 mM. it does not show differences. This work had several difficulties, however, important results were obtained that can lead to more detailed studies of the oxidation-reduction process during SC. Key words: glutathione, sperm capacitation, in vitro fertilization, reactive oxygen species. |
|---|