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L’intégrine α6β1 ovocytaire et spermatique dans l’interaction gamétique

Sperm and oocyte α6β1 integrin in gamete interaction

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Résumé

Des tests d’inhibitions ont permis de proposer l’intégrine α6β1 comme le récepteur du spermatozoïde sur l’ovocyte mais des expériences d’invalidation de gène ont montré que les sous unités intégrines α6 et β1 ovocytaires n’étaient pas essentielles à la fécondation. En utilisant le Western blot et l’immunofluorescence (cytométrie de flux et microscopie), nous avons montré que I’α6β1 est exprimée par les spermatozoïdes de souris. Comme pour l’ovocyte, un anticorps anti-α6 (GoH3) inhibe spécifiquement les capacités fécondantes des spermatozoïdes.

En comparant des tests de fusion avec des ovocytes intacts (avec zone pellucide) ou des ovocytes dépellucidés, nous avons montré que la dépellucidation court-circuite la fonction de l’α6β1 dans le processus d’adhésion/fusion des gamètes. L’α6β1 est donc exprimée et fonctionnelle sur les deux gamètes au cours de l’interaction de leur membranes.

Nos résultats, ceux des expériences utilisant des ovocytes mutés systématiquement inséminés avec des spermatozoïdes de souris sauvages, ainsi que ceux des myoblastes qui sont incapables de fusionner entre eux lorsqu’ils sont mutés pour la sous-unité intégrine β1 alors qu’ils le font avec des myoblastes sauvages, nous conduisent à formuler l’hypothèse selon laquelle la présence de la sous-unité intégrine β1 sur une seule des deux membranes suffit pour que la fusion ait lieu. Cette hypothèse est renforcée par un phénomène d’échange de fragments membranaires entre les gamètes juste avant leur fusion que nous avons décrits tout récemment.

Abstract

Based on inhibition tests, the α6β1 integrin was suggested to be a sperm receptor, but further experiments using gene deletion techniques have shown that neither oocyte α6, nor β1 integrin subunits were essential for mouse fertilization. Using Western blot analysis and immunofluorescence (flow cytometry and microscopy), we have shown that mouse sperm expresses the α6β1 integrin.

As for oocytes, binding of GoH3 anti-alpha6 antibody to sperm induces specific inhibition of sperm fertilizing ability. Comparing zona-intact and zona-free eggs in fusion tests, we have shown that removal of the zona pellucida bypasses the α6β1 integrin role in the adhesion/fusion process of oocyte fertilization. The α6β1 integrin is expressed by both gametes and is functional during their membrane interactions.

Our results, previous reports on fertilization of α6 or β1 integrin subunit-deleted oocytes by wild-type sperm and the fusion ability of β1 mutant myoblasts when they were co-cultured with wild-type myoblasts suggest that the presence of α6βl integrin on one of the two gamete membranes can rescue the fertilization process. This hypothesis is further supported by the recently reported exchange of membrane fragments occurring between gametes prior to fusion.

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Correspondence to Ahmed Ziyyat or Virginie Barraud-Lange or Jean -Philippe Wolf.

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Ziyyat, A., Barraud-Lange, V. & Wolf, J.-. L’intégrine α6β1 ovocytaire et spermatique dans l’interaction gamétique. Androl. 18, 122–126 (2008) doi:10.1007/BF03040389

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Mots clés

  • fécondation
  • spermatozoïde
  • ovocyte
  • intégrine

Key words

  • fertilization
  • sperm
  • oocyte
  • integrin
  • zona pellucida