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Les larges vacuoles des têtes spermatiques sont-elles associées à des altérations du noyau ou de l’acrosome du spermatozoïde ?

Is there a relationship between large vacuoles in spermatozoa and nucleus or acrosome alterations?

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Résumé

Objectifs

Rechercher la présence d’altérations nucléaires ou acrosomiques dans les spermatozoïdes présentant des vacuoles occupant au moins 13 % de la surface de la tête des spermatozoïdes (spermatozoïdes dits de type 3), détectées à l’aide de la technique MSOME (motile sperm organelle morphology examination).

Matériel et méthodes

Notre étude a été effectuée sur des recueils de sperme de 20 hommes infertiles. Une évaluation de l’acrosome par immunomarquage à l’aide d’un anticorps monoclonal antiproacrosine (Mab4D4) a été associée à une détection de la fragmentation de l’ADN par la technique TUNEL. La condensation de la chromatine spermatique a été évaluée par une coloration au bleu d’aniline, et le taux d’aneuploïdie a été déterminé par hybridation in situ en fluorescence (FISH). Une observation des échantillons spermatiques par microscopie électronique à transmission (MET) [sperme entier] et confocale [spermatozoïdes de type 3 isolés] a complété ces analyses. Les résultats ont été comparés entre des spermatozoïdes provenant du sperme entier et des spermatozoïdes de type 3 isolés.

Résultats

Une augmentation significative de la proportion d’acrosomes anormaux a été constatée dans les spermatozoïdes isolés de type 3 par comparaison au sperme entier (77,8 ± 2,49 % vs 70,6 ± 2,62 %). La fragmentation de l’ADN a été observée de manière comparable dans les spermatozoïdes de type 3 et le sperme entier (14,5 ± 3,45 % vs 11,5 ± 1,25%), un défaut de condensation de la chromatine ainsi qu’une augmentation significative du taux d’aneuploïdie ont été retrouvés dans les spermatozoïdes isolés par comparaison à ceux provenant du sperme entier (50,4 ± 3,10 % vs 26,5 ± 2,60 % et 7,8 ± 1,98 % vs 1,3 ± 0,18 %). Les larges vacuoles sont à localisation nucléaire exclusive (MET et microscopie confocale).

Conclusion

Les vacuoles seraient en relation directe avec un défaut de maturation nucléaire survenant au cours de la spermiogenèse.

Abstract

Objective

The aim of this study was to detect acrosome and nucleus alterations in isolated spermatozoa with large vacuoles detected by MSOME (Motile Sperm Organelle Morphology Examination), named type 3 spermatozoa and defined by the presence of one or more vacuoles occupying more than 13% of the sperm head area.

Material and methods

Twenty infertile men were included in this study. Whole sperm and isolated spermatozoa were compared. Spermatozoa acrosome and nucleus were explored using 1) proacrosin immunostaining with a monoclonal antibody (4D4), 2) DNA fragmentation with TUNEL assay, 3) chromatin condensation with aniline blue staining, and 4) aneuploidy after fluorescence in situ hybridization (FISH) and analysis by electron transmission and confocal microscopy.

Results

Acrosome abnormalities were significantly increased in type 3 spermatozoa compared towhole sperm(77.8 ± 2.49% vs. 70.6 ± 2.62%). DNA fragmentation was similar in type 3 spermatozoa compared towhole sperm(14.5 ± 3.45%vs. 11.5 ± 1.25%). Chromatin condensation was significantly altered in isolated spermatozoa as well as aneuploidy frequencies (50.4 ± 3.10% vs. 26.5 ± 2.60% and 7.8 ± 1.98% vs. 1.3 ± 0.18%). Large vacuoles have an exclusive nuclear location, confirmed by electron and confocal microscopy.

Conclusion

Large vacuoles are probably due to sperm nucleus maturation dysfunction during spermiogenesis.

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Correspondence to N. Rives.

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Travers, A., Perdrix, A., Legrand, F. et al. Les larges vacuoles des têtes spermatiques sont-elles associées à des altérations du noyau ou de l’acrosome du spermatozoïde ?. Basic Clin. Androl. 20, 247–256 (2010) doi:10.1007/s12610-010-0091-2

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Mots clés

  • Aneuploïdie
  • Chromatine
  • MSOME
  • Spermatozoïdes
  • Vacuoles

Keywords

  • Aneuploidy
  • Chromatin
  • MSOME
  • Spermatozoa
  • Vacuoles