Evaluation of the human sperm nucleus: ambiguity and risk of confusion with chromomycin staining

Ménézo, Yves

doi:10.1017/s0967199420000672

Cited by 4 publications

(2 citation statements)

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“…However, CMA3 has proven helpful for assessing pig spermatozoa in some situations [39], and it could be applied as a way to discard suboptimal males when they enter stud centers or to evaluate the impact of conditions such as heat stress [54] Additionally, CMA3 interpretation is controversial, since studies in human have shown a striking coincidence of DNA oxidation, SDF, and CMA3 results [34]. Indeed, CMA3 cannot reflect the degree of protamination [25], and SDF could modulate the interaction of CMA3 with DNA, altering the evaluation of chromatin compaction by this technique. However, we remain ignorant of whether the situation could be the same for pigs, considering the differences in chromatin structure for both species (including the presence of two protamines in human sperm).…”

Section: Cma3 For the Evaluation Of Chromatin Compaction In Pig Sperm...mentioning

confidence: 99%

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Application of Flow Cytometry Using Advanced Chromatin Analyses for Assessing Changes in Sperm Structure and DNA Integrity in a Porcine Model

Lacalle,

Fernández-Alegre,

Gómez-Giménez

et al. 2024

IJMS

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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.

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Section: Cma3 For the Evaluation Of Chromatin Compaction In Pig Sperm...mentioning

confidence: 99%

“…Widely used in humans, high CMA3 labeling has been associated with lower male fertility [24]. Nevertheless, the interpretation of the CMA3 assay is complex since it is affected by other factors, such as DNA fragmentation and the overall quantity of disulfide bridges among protamines [25].…”

Section: Introductionmentioning

confidence: 99%