Caracterización de proteínas conjugadas a ubiquitina en espermatozoides humanos capacitados
Capacitation is a set of biochemical and physiological changes that occur in spermatozoa during their transit along the female reproductive tract that are necessary to become cells able to fertilize the female gamete. Although several mechanisms related to capacitation have been studied, such as the...
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| Format: | master thesis |
| Status: | Published version |
| Publication Date: | 2022 |
| Country: | México |
| Institution: | Universidad Autónoma Metropolitana |
| Repository: | Repositorio Institucional de la UAM Iztapalapa |
| Language: | Spanish |
| OAI Identifier: | oai:bindani.izt.uam.mx:vh53wv951 |
| Online Access: | https://doi.org/10.24275/uami.vh53wv951 |
| Access Level: | Open access |
| Keyword: | info:eu-repo/classification/LEM/Fecundación info:eu-repo/classification/LEM/Espermatozoides info:eu-repo/classification/LEM/Spermatozoa info:eu-repo/classification/LEM/Espermatogénesis info:eu-repo/classification/LEM/Spermatogenesis info:eu-repo/classification/LEM/Fertilization (Biology) info:eu-repo/classification/cti/6 |
| Summary: | Capacitation is a set of biochemical and physiological changes that occur in spermatozoa during their transit along the female reproductive tract that are necessary to become cells able to fertilize the female gamete. Although several mechanisms related to capacitation have been studied, such as the increase of tyrosine phosphorylation, there are still some others unknown. One of the mechanisms involved in capacitation that is poorly studied is the ubiquitinproteasome system. The aim of this project was to characterize the proteins conjugated to ubiquitin in human sperm during the in vitro capacitation. To that purpose, we studied the changes in ubiquitinated proteins of non-capacitated and capacitated human spermatozoa by Western blot, after protein separation by denaturing electrophoresis (SDS-PAGE) and two-dimensional electrophoresis (2DE). Moreover, the proteins conjugated to ubiquitin were isolated by immunoprecipitation. The results show the increase of protein ubiquitin conjugation during capacitation, simultaneous to the increase of tyrosine phosphorylation. Further analysis of the ubiquitin conjugates allowed the identification of 6 protein groups differentially ubiquitinated. When the proteasome activity was inhibited, we found the accumulation of ubiquitin conjugates, while the tyrosine phosphorylation remained steady. The immunoprecipitation allowed the isolation of ubiquitin conjugates, but the recovery was very low. The results support that the sperm proteins conjugated to ubiquitin and their degradation by the proteasome participate during capacitation, so their study and identification may contribute to the better knowledge of the signaling pathways necessary for sperm to fertilize. |
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