Caracterización de glicoproteínas y colesterol de los microdominios de la membrana de espermatozoides de cerdo durante la capacitación y criopreservación
The surface of the spermatozoa is coated with glycoproteins, the redistribution of which during in vitro capacitation plays a vital role in the subsequent fertilization process. Lipid rafts are membrane microdomains involved in signal transduction through receptors and include or recruit specific ty...
| Autor: | |
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| Tipo de recurso: | tesis doctoral |
| Estado: | Versión publicada |
| Fecha de publicación: | 2021 |
| País: | México |
| Institución: | Universidad Autónoma Metropolitana |
| Repositorio: | Repositorio Institucional de la UAM Iztapalapa |
| Idioma: | español |
| OAI Identifier: | oai:bindani.izt.uam.mx:6h440s70n |
| Acceso en línea: | https://doi.org/10.24275/uami.6h440s70n |
| Access Level: | acceso abierto |
| Palabra clave: | info:eu-repo/classification/LEM/Cerdos -- Reproducción info:eu-repo/classification/LEM/Tecnología reproductiva info:eu-repo/classification/LEM/Capacitación espermática info:eu-repo/classification/LEM/Swine -- Reproduction info:eu-repo/classification/LEM/Reproductive technology info:eu-repo/classification/LEM/Spermatozoa info:eu-repo/classification/cti/3 |
| Sumario: | The surface of the spermatozoa is coated with glycoproteins, the redistribution of which during in vitro capacitation plays a vital role in the subsequent fertilization process. Lipid rafts are membrane microdomains involved in signal transduction through receptors and include or recruit specific types of proteins and glycoproteins. Few studies have focused on identifying glycoproteins resident in the lipid rafts of spermatozoa. Proteins associated with lipid rafts modify their localization during capacitation. The study's objective was to identify the glycoproteins associated with lipid rafts of capacitated boar spermatozoa through a lectin-binding assay coupled to a mass spectrometry approach. From the proteomic profiles generated by the raft proteins extractions, we observed that some bands' intensity increased after capacitation while others' decreased. To determine whether the proteins obtained from lipid rafts are glycosylated, lectin blot assays were performed, the protein bands with good resolution and significant glycosylation modifications after capacitation were analyzed by mass spectrometry. The bands of interest had an apparent molecular weight of 64, 45, 36, 34, 24, 18, and 15 kDa. We sequenced the seven bands, and 20 known or potential glycoproteins were identified. In our knowledge, this is the first time that the following proteins and their association with sperm lipid rafts are described: ADAM5, SPMI, SPACA1, Seminal plasma protein pB1, PSP-I, MFGE8, tACE, PGK2, SUCLA2, and MDH1. Moreover, LYDP4, SPAM-1, HSP60, ZPBP1, and AK1 were previously reported in mouse and human spermatozoa lipid rafts but not in boar spermatozoa. We also found and confirmed the presence of ACR, ACRBP, AWN, AQN3, and PRDX5 in lipid rafts of boar spermatozoa. This paper provides an overview of the glycosylation pattern in lipid rafts of boar spermatozoa before and after capacitation. The further glycomic analysis is needed to determine the type and the variation of glycan chains of the lipid rafts glycoproteins on the surface of spermatozoa during capacitation and acrosome reaction. Five patterns of cholesterol distribution of the rafts were found in non- capacitated sperm: Pattern A with fluorescence in the flagellum and head region predominated; after capacitation, it was pattern C with fluorescence in the head, in cryopreserved two patterns were observed more being the majority patterns A and C. Regarding the concentration of cholesterol, it was found that in cryopreserved sperm there was considerable loss of cholesterol which was detected in the freezing medium, so it is postulated that the freezing process affected the sperm membrane considerably. |
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