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First published online 17 January 2006
doi: 10.1242/jcs.02730

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CD9 controls the formation of clusters that contain tetraspanins and the integrin 6ß1, which are involved in human and mouse gamete fusion

Ahmed Ziyyat^1,*, Eric Rubinstein^2,*, Frédérique Monier-Gavelle¹, Virginie Barraud¹, Olivier Kulski¹, Michel Prenant², Claude Boucheix², Morgane Bomsel³ and Jean-Philippe Wolf^1,

¹ Université Paris 13, Laboratoire de Biologie de la Reproduction, UPRES 3410, UFR SMBH, Bobigny, France; AP-HP, Hôpital Jean Verdier, Service d'Histologie, Embryologie et Cytogénétique, Bondy, France
² INSERM, U602, Villejuif, France; Université Paris XI, Villejuif, France; Institut André Lwoff, Villejuif, France
³ INSERM, U567, Institut Cochin, Paris, France

Author for correspondence (e-mail: jean-philippe.wolf{at}jvr.ap-hop-paris.fr)

Accepted 19 September 2005

The process of gamete fusion has been largely studied in the mouse and has revealed the crucial role of the tetraspanin CD9. By contrast, human gamete fusion remains largely unknown. We now show that an anti-6 integrin mAb (GoH3) strongly inhibited human sperm-egg fusion in human zona-free eggs. Furthermore, a mAb directed against CD151, a tetraspanin known to associate with 6ß1, partially inhibited sperm-egg fusion. By contrast, the addition of an anti-CD9 mAb to zona free eggs had no effect. The integrin 6ß1, CD151 and CD9 tetraspanins were evenly distributed on human zona-intact oocytes. On zona-free eggs, the integrin 6ß1 and tetraspanin CD151 patched and co-localized but the tetraspanin CD9 remained unchanged. CD9 mAb prevented 6ß1 integrin clustering and gamete fusion when added prior to, but not after, zona removal. Antibody-mediated aggregation of integrin 6ß1 yielded patches that were bigger and more heterogeneous in mouse oocytes lacking CD9. Moreover, a strong labelling of 6ß1 could be observed at the sperm entry point. Altogether, these data show that CD9 controls the redistribution of some membrane proteins including the 6ß1 integrin into clusters that may be necessary for gamete fusion.

Key words: Human, Fertilization, Integrins, Tetraspanins, CD9, CD151

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