- Article Original
- Intéraction Gamétique
- Published:
L’intégrine α6β1 ovocytaire et spermatique dans l’interaction gamétique
Sperm and oocyte α6β1 integrin in gamete interaction
Andrologie volume 18, pages 122–126 (2008)
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.
References
ALMEIDA E.A., HUOVILA A.R, SUTHERLAND A.E. et al.: Mouse egg integrin alpha 6 beta 1 functions as a sperm receptor. Cell, 1995, 81: 1095–10104.
BARRAUD-LANGE V., NAUD-BARRIANT N., BOMSEL M., WOLF J.P., ZIYYAT A.: Transfer of oocyte membrane fragments to fertilizing spermatozoa. Faseb J., 2007, 21: 3446–3449.
BARRAUD-LANGE V., NAUD-BARRIANT N., SAFFAR L. et al.: Alpha6beta1 integrin expressed by sperm is determinant in mouse fertilization. BMC Dev. Biol., 2007, 7: 102.
BIGLER D., TAKAHASHI Y., CHEN M.S., ALMEIDA E.A., OSBOURNE L., WHITE J.M.: Sequence-specific interaction between the disintegrin domain of mouse ADAM 2 (fertilin beta) and murine eggs. Role of the alpha(6) integrin subunit. J. Biol. Chem., 2000, 275: 11576–11584.
CHEN H., SAMPSON N.S.: Mediation of sperm-egg fusion: evidence that mouse egg alpha6beta1 integrin is the receptor for sperm fertilinbeta. Chem. Biol., 1999, 6 :1–10.
CHEN M.S., TUNG K.S., COONROD S.A. et al.: Role of the integrin-associated protein CD9 in binding between sperm ADAM 2 and the egg integrin alpha6beta1: implications for murine fertilization. Proc. Natl Acad. Sei. USA, 1999, 96: 11830–11835.
CHO C, BUNCH D.O., FAURE J.E. et al.: Fertilization defects in sperm from mice lacking fertilin beta. Science, 1998, 281: 1857–1859.
COX S.L., SHAW J., JENKIN G.: Transplantation of cryopreserved fetal ovarian tissue to adult recipients in mice. J. Reprod. Fertil., 1996, 107: 315–322.
EVANS J.P.: The molecular basis of sperm-oocyte membrane interactions during mammalian fertilization. Hum. Reprod. Update, 2002, 8: 297–311.
EVANS J.P., SCHULTZ R.M., KOPF G.S.: Identification and localization of integrin subunits in oocytes and eggs of the mouse. Mol. Reprod. Dev., 1995, 40: 211–220.
FUSI F.M., VIGNALI M., BUSACCA M., BRONSON R.A.: Evidence for the presence of an integrin cell adhesion receptor on the oolemma of unfertilized human oocytes. Mol. Reprod. Dev., 1992, 31: 215–222.
GLANDER HJ., SCHALLER J.: Beta 1-integrins of spermatozoa: a flow cytophotometric analysis. Int. J. Androl., 1993, 16: 105–111.
GLANDER HJ., SCHALLER J., WEBER W., ALEXANDER H., HAAKE K.W.: In vitro fertilization: increased VLA (very late antigen) integrins and fibronectin after acrosome reaction. Arch. Androl., 1996, 36: 177–185.
HE Z.Y., BRAKEBUSCH C, FASSLER R., KREIDBERG J.A., PRIMAKOFF P., MYLES D.G.: None of the integrins known to be present on the mouse egg or to be ADAM receptors are essential for sperm-egg binding and fusion. Dev. Biol., 2003, 254: 226–237.
HEMLER M.E.: Integrin associated proteins. Curr. Opin. Cell. Biol., 1998, 10: 578–585.
HENKEL R., SCHALLER J., GLANDER H.J., SCHILL W.B.: Low expression of adhesion molecules and matrix proteins in patients showing poor penetration in zona-free hamster oocytes. Mol. Hum. Reprod., 1996, 2: 335–339.
HIERCK B.P., THORSTEINSDOTTIR S., NIESSEN CM. et al.: Variants of the alpha 6 beta 1 laminin receptor in early murine development: distribution, molecular cloning and chromosomal localization of the mouse integrin alpha 6 subunit. Cell Adhes. Commun., 1993, 1: 33–53.
INOUE N., IKAWA M., ISOTANI A., OKABE M.: The immunoglobulin superfamily protein Izumo is required for sperm to fuse with eggs. Nature, 2005, 434: 234–238.
JI Y.Z., WOLF J.P., JOUANNET P., BOMSEL M.: Human gamete fusion can bypass beta1 integrin requirement. Hum. Reprod., 1998, 13: 682–689.
KAJI K., ODA S., SHIKANO T. et al.: The gamete fusion process is defective in eggs of Cd9-deficient mice. Nat. Genet., 2000, 24: 279–282.
KLENTZERIS L.D., FISHEL S., MCDERMOTT H., DOWELL K., HALL J., GREEN S.: A positive correlation between expression of beta 1-integrin cell adhesion molecules and fertilizing ability of human spermatozoa in vitro. Hum. Reprod., 1995, 10: 728–733.
LE NAOUR F., RUBINSTEIN E., JASMIN C., PRENANT M., BOUCHEIX C.: Severely reduced female fertility in CD9-deficient mice. Science, 2000, 287: 319–321.
MILLER B.J., GEORGES-LABOUESSE E., PRIMAKOFF P., MYLES D.G.: Normal fertilization occurs with eggs lacking the integrin alpha6beta1 and is CD9-dependent. J. Cell. Biol., 2000, 149: 1289–1296.
MIYADO K., YAMADA G., YAMADA S. et al.: Requirement of CD9 on the egg plasma membrane for fertilization. Science, 2000, 287: 321–324.
REDDY K.V., MEHERJI P.K., SHAHANI S.K.: Integrin cell adhesion molecules on human spermatozoa. Indian J. Exp. Biol., 1998, 36: 456–463.
REDDY V.R., RAJEEV S.K., GUPTA V.: Alpha 6 beta 1 Integrin is a potential clinical marker for evaluating sperm quality in men. Fertil. Steril., 2003, 79 Suppl 3:1590–1596.
RUBINSTEIN E., LE NAOUR F., LAGAUDRIERE-GESBERT C., BILLARD M., CONJEAUD H., BOUCHEIX C.: CD9, CD63, CD81, and CD82 are components of a surface tetraspan network connected to HLA-DR and VLAintegrins. Eur. J. Immunol., 1996, 26: 2657–2665.
RUBINSTEIN E., POINDESSOUS-JAZAT V, LE NAOUR F., BILLARD M., BOUCHEIX C.: CD9, but not other tetraspans, associates with the betai integrin precursor. Eur. J. Immunol., 1997, 27: 1919–1927.
RUNGE K.E., EVANS J.E., HE Z.Y. et al.: Oocyte CD9 is enriched on the microvillar membrane and required for normal microvillar shape and distribution. Dev. Biol., 2007, 304: 317–325.
SCHWANDER M., LEU M., STUMM M. ét al.: Betai integrins regulate myoblast fusion and sarcomere assembly. Dev. Cell, 2003, 4: 673–685.
SNELL W.J., WHITE J.M.:The molecules of mammalian fertilization. Cell, 1996, 85: 629–637.
TAKAHASHI Y., BIGLER D., ITO Y., WHITE J.M.: Sequence-specific interaction between the disintegrin domain of mouse ADAM 3 and murine eggs: role of beta1 integrin-associated proteins CD9, CD81, and CD98. Mol. Biol. Cell, 2001, 12: 809–820.
TAKAHASHI Y, YAMAKAWA N., MATSUMOTO K., TOYODA Y., FURUKAWA K., SATO E.: Analysis of the role of egg integrins in sperm-egg binding and fusion. Mol. Reprod. Dev., 2000, 56: 412–423.
TARONE G., RUSSO M.A., HIRSCH E. et al.: Expression of beta 1 integrin complexes on the surface of unfertilized mouse oocyte. Development, 1993, 117: 1369–1375.
YANAGIMACHI R.: Fertility of mammalian spermatozoa: its development and relativity. Zygote, 1994, 2: 371–372
ZIYYAT A., RUBINSTEIN E., MONIER-GAVELLE F. et al.: CD9 controls the formation of clusters that contain tetraspanins and the integrin alpha6beta1, which are involved in human and mouse gamete fusion. J. Cell. Sci., 2006, 119: 416–424.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Ziyyat, A., Barraud-Lange, V. & Wolf, J.P. L’intégrine α6β1 ovocytaire et spermatique dans l’interaction gamétique. Androl. 18, 122–126 (2008). https://doi.org/10.1007/BF03040389
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF03040389