Previous results, based on inhibition of fertilization by an anti–α6 integrin mAb (GoH3), suggest that the α6β1 integrin on mouse eggs functions as the receptor for sperm (Almeida, E.A., A.P. Huovila, A.E. Sutherland, L.E. Stephens, P.G. Calarco, L.M. Shaw, A.M. Mercurio, A. Sonnenberg, P. Primakoff, D.G. Myles, and J.M. White. 1995. Cell. 81:1095–1104). Because the egg surface tetraspanin CD9 is essential for gamete fusion (Kaji, K., S. Oda, T. Shikano, T. Ohnuki, Y. Uematsu, J. Sakagami, N. Tada, S. Miyazaki, and A. Kudo. 2000. Nat. Genet. 24:279–282; Le Naour, F., E. Rubinstein, C. Jasmin, M. Prenant, and C. Boucheix. 2000. Science. 287:319–321; Miyado, K., G. Yamada, S. Yamada, H. Hasuwa, Y. Nakamura, F. Ryu, K. Suzuki, K. Kosai, K. Inoue, A. Ogura, M. Okabe, and E. Mekada. 2000. Science. 287:321–324) and CD9 is known to associate with integrins, recent models of gamete fusion have posited that egg CD9 acts in association with α6β1 in fusion (Chen, M.S., K.S. Tung, S.A. Coonrod, Y. Takahashi, D. Bigler, A. Chang, Y. Yamashita, P.W. Kincade, J.C. Herr, and J.M. White. 1999. Proc. Natl. Acad. Sci. USA. 96:11830–11835; Kaji, K., S. Oda, T. Shikano, T. Ohnuki, Y. Uematsu, J. Sakagami, N. Tada, S. Miyazaki, and A. Kudo. 2000. Nat. Genet. 24:279–282; Le Naour, F., E. Rubinstein, C. Jasmin, M. Prenant, and C. Boucheix. 2000. Science. 287:319–321; Miyado, K., G. Yamada, S. Yamada, H. Hasuwa, Y. Nakamura, F. Ryu, K. Su- zuki, K. Kosai, K. Inoue, A. Ogura, M. Okabe, and E. Mekada. 2000. Science. 287:321–324). Using eggs from cultured ovaries of mice lacking the α6 integrin subunit, we found that the fertilization rate, fertilization index, and sperm binding were not impaired compared with wild-type or heterozygous controls. Furthermore, a reexamination of antibody inhibition, using an assay that better simulates in vivo fertilization conditions, revealed no inhibition of fusion by the GoH3 mAb. We also found that an anti-CD9 mAb completely blocks sperm fusion with either wild-type eggs or eggs lacking α6β1. Based on these results, we conclude that the α6β1 integrin is not essential for sperm–egg fusion, and we suggest a new model in which CD9 acts by itself, or interacts with egg protein(s) other than α6β1, to function in sperm–egg fusion.
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12 June 2000
Article|
June 12 2000
Normal Fertilization Occurs with Eggs Lacking the Integrin α6β1 and Is Cd9-Dependent
Brent J. Miller,
Brent J. Miller
aDepartment of Molecular and Cellular Biology, University of California Davis, Davis, California 95616
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Elisabeth Georges-Labouesse,
Elisabeth Georges-Labouesse
cInstitut de Génétique et de Biologie Moléculaire et Cellulaire, Centre National de la Recherche Scientifique, Institut National de la Sante et de la Recherche Medicale, ULP, 67404 Illkirch, C.U. de Strasbourg, France
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Paul Primakoff,
Paul Primakoff
bDepartment of Cell Biology and Human Anatomy, School of Medicine, University of California Davis, Davis, California 95616
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Diana G. Myles
Diana G. Myles
aDepartment of Molecular and Cellular Biology, University of California Davis, Davis, California 95616
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Brent J. Miller
aDepartment of Molecular and Cellular Biology, University of California Davis, Davis, California 95616
Elisabeth Georges-Labouesse
cInstitut de Génétique et de Biologie Moléculaire et Cellulaire, Centre National de la Recherche Scientifique, Institut National de la Sante et de la Recherche Medicale, ULP, 67404 Illkirch, C.U. de Strasbourg, France
Paul Primakoff
bDepartment of Cell Biology and Human Anatomy, School of Medicine, University of California Davis, Davis, California 95616
Diana G. Myles
aDepartment of Molecular and Cellular Biology, University of California Davis, Davis, California 95616
Abbreviations used in this paper: DAPI, 4′,6′-diamidino-2-phenylindole dihydrochloride; FI, fertilization index; FR, fertilization rate.
Received:
April 10 2000
Revision Requested:
May 09 2000
Accepted:
May 10 2000
Online ISSN: 1540-8140
Print ISSN: 0021-9525
© 2000 The Rockefeller University Press
2000
The Rockefeller University Press
J Cell Biol (2000) 149 (6): 1289–1296.
Article history
Received:
April 10 2000
Revision Requested:
May 09 2000
Accepted:
May 10 2000
Connected Content
Citation
Brent J. Miller, Elisabeth Georges-Labouesse, Paul Primakoff, Diana G. Myles; Normal Fertilization Occurs with Eggs Lacking the Integrin α6β1 and Is Cd9-Dependent. J Cell Biol 12 June 2000; 149 (6): 1289–1296. doi: https://doi.org/10.1083/jcb.149.6.1289
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