Compartir
Título
Application of Flow Cytometry Using Advanced Chromatin Analyses for Assessing Changes in Sperm Structure and DNA Integrity in a Porcine Model
Autor
Facultad/Centro
Área de conocimiento
Título de la revista
International Journal of Molecular Sciences
Número de la revista
4
Datos de la obra
Lacalle, E., Fernández-Alegre, E., Gómez-Giménez, B., Álvarez-Rodríguez, M., Martín-Fernández, B., Soriano-Úbeda, C. and Martínez-Pastor, F. (2024). Application of flow cytometry using advanced chromatin analyses for assessing changes in sperm structure and DNA integrity in a porcine model. International Journal of Molecular Sciences, 25(4), Article e1953. https://doi.org/10.3390/ijms25041953
Editor
MDPI
Fecha
2024
ISSN
1661-6596
Abstract
[EN] Chromatin status is critical for sperm fertility and reflects spermatogenic success. We tested a multivariate approach for studying pig sperm chromatin structure to capture its complexity with a set of quick and simple techniques, going beyond the usual assessment of DNA damage. Sperm doses from 36 boars (3 ejaculates/boar) were stored at 17 °C and analyzed on days 0 and 11. Analyses were: CASA (motility) and flow cytometry to assess sperm functionality and chromatin structure by SCSA (%DFI, DNA fragmentation; %HDS, chromatin maturity), monobromobimane (mBBr, tiol status/disulfide bridges between protamines), chromomycin A3 (CMA3, protamination), and 8-hydroxy-2′-deoxyguanosine (8-oxo-dG, DNA oxidative damage). Data were analyzed using linear models for the effects of boar and storage, correlations, and multivariate analysis as hierarchical clustering and principal component analysis (PCA). Storage reduced sperm quality parameters, mainly motility, with no critical oxidative stress increases, while chromatin status worsened slightly (%DFI and 8-oxo-dG increased while mBBr MFI—median fluorescence intensity—and disulfide bridge levels decreased). Boar significantly affected most chromatin variables except CMA3; storage also affected most variables except %HDS. At day 0, sperm chromatin variables clustered closely, except for CMA3, and %HDS and 8-oxo-dG correlated with many variables (notably, mBBr). After storage, the relation between %HDS and 8-oxo-dG remained, but correlations among other variables disappeared, and mBBr variables clustered separately. The PCA suggested a considerable influence of mBBr on sample variance, especially regarding storage, with SCSA and 8-oxo-dG affecting between-sample variability. Overall, CMA3 was the least informative, in contrast with results in other species. The combination of DNA fragmentation, DNA oxidation, chromatin compaction, and tiol status seems a good candidate for obtaining a complete picture of pig sperm nucleus status. It raises many questions for future molecular studies and deserves further research to establish its usefulness as a fertility predictor in multivariate models. The usefulness of CMA3 should be clarified
Materia
Palabras clave
Peer review
SI
ID proyecto
- info:eu-repo/grantAgreement/AEI/Programa Estatal de I+D+i Orientada a los Retos de la Sociedad/ RTI2018-095183-B-I00/Incremento de la eficiencia reproductiva de la cría porcina mediante la aplicación de técnicas avanzadas de análisis de la cromatina espermática y selección por coloide/
- info:eu-repo/grantAgreement/Junta de Castilla y León/Programa de apoyo a proyectos de investigación, cofinanciadas por el Fondo Europeo de Desarrollo Regional/LE023P20/Mejora de la eficiencia de la industria porcina mediante la aplicación de técnicas avanzadas de selección espermática y análisis de la cromatina espermática
- info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/DIN2020-011558/
URI
DOI
Versión del editor
Collections
- Untitled [5431]
Files in this item
Tamaño:
1.243
xmlui.dri2xhtml.METS-1.0.size-megabytes
Formato:
Adobe PDF