RT info:eu-repo/semantics/article
T1 Application of Flow Cytometry Using Advanced Chromatin Analyses for Assessing Changes in Sperm Structure and DNA Integrity in a Porcine Model
A1 Lacalle Fernández, Estíbaliz
A1 Fernández Alegre, Estela
A1 Gómez Giménez, Belén
A1 Álvarez Rodríguez, Manuel
A1 Martín Fernández, Beatriz
A1 Soriano Úbeda, Cristina de las Mercedes
A1 Martínez Pastor, Felipe
A2 Biologia Celular
K1 Biología
K1 Veterinaria
K1 Boar
K1 Spermatozoa
K1 Cooled storage
K1 Flow cytometry
K1 Chromatin status
K1 Oxidative stress
K1 DNA fragmentation
K1 Sperm functionality
K1 Multivariate analysis
K1 3104.11 Reproducción
K1 3104.08 Porcinos
K1 2407.02 Citogenética
AB [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
PB MDPI
SN 1661-6596
LK https://hdl.handle.net/10612/20354
UL https://hdl.handle.net/10612/20354
NO 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
NO This article belongs to the Special Issue Novel Insights into the Biology of Spermatozoa 2.0
DS BULERIA. Repositorio Institucional de la Universidad de León
RD 26-jun-2024