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The Sperm‐surface glycoprotein, SGP, is necessary for fertilization in the frog, Xenopus laevis
Author(s) -
Nagai Keita,
Ishida Takuya,
Hashimoto Takafumi,
Harada Yuichirou,
Ueno Shuichi,
Ueda Yasushi,
Kubo Hideo,
Iwao Yasuhiro
Publication year - 2009
Publication title -
development, growth and differentiation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.864
H-Index - 66
eISSN - 1440-169X
pISSN - 0012-1592
DOI - 10.1111/j.1440-169x.2009.01112.x
Subject(s) - xenopus , sperm , human fertilization , biology , glycoprotein , microbiology and biotechnology , salientia , vitelline membrane , vesicle , monoclonal antibody , embryo , antibody , biochemistry , oocyte , membrane , anatomy , immunology , genetics , gene
To identify a molecule involved in sperm‐egg plasma membrane binding at fertilization, a monoclonal antibody against a sperm‐surface glycoprotein (SGP) was obtained by immunizing mice with a sperm membrane fraction of the frog, Xenopus laevis , followed by screening of the culture supernatants based on their inhibitory activity against fertilization. The fertilization of both jellied and denuded eggs was effectively inhibited by pretreatment of sperm with intact anti‐SGP antibody as well as its Fab fragment, indicating that the antibody recognizes a molecule on the sperm's surface that is necessary for fertilization. On Western blots, the anti‐SGP antibody recognized large molecules, with molecular masses of 65–150 kDa and minor smaller molecules with masses of 20–28 kDa in the sperm membrane vesicles. SGP was distributed over nearly the entire surface of the sperm, probably as an integral membrane protein in close association with microfilaments. More membrane vesicles containing SGP bound to the surface were found in the animal hemisphere compared with the vegetal hemisphere in unfertilized eggs, but the vesicle‐binding was not observed in fertilized eggs. These results indicate that SGP mediates sperm‐egg membrane binding and is responsible for the establishment of fertilization in Xenopus .