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J. Biol. Chem., Vol. 280, Issue 27, 25928-25935, July 8, 2005
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From the Department of Cell Biology and Human Anatomy, School of Medicine, University of California, Davis, California 95616
We demonstrated previously the involvement of a nicotinic acetylcholine
receptor containing an 
7 subunit in the human sperm acrosome reaction
(a modified exocytotic event essential to fertilization). Here we report the
presence in human sperm of 7, 9, 3, 5, and 
4
nicotinic acetylcholine receptor subunits and the following proteins known to
be associated with the receptor in the somatic cell: rapsyn and the tyrosine
kinases c-SRC and FYN. The 7 subunit appears to exist as a homomer in
the posterior post-acrosomal and neck regions of sperm and is probably linked
to the cytoskeleton via rapsyn. The 3, 5, and 4 subunits
are present in the sperm flagellar mid-piece of sperm and possibly exist as
354 and/or 34 channels. The 9 subunit
is present in the sperm mid-piece. We detected the FYN and c-SRC tyrosine
kinases in the flagellar mid-piece region. Both co-precipitated only with the
nicotinic acetylcholine receptor 4 subunit. Immunolocalization with a
C-terminal SRC kinase antibody, which recognizes several members of SRC kinase
family, detected a SRC kinase co-localized with the 7 subunit in the
neck region of sperm. Immunoprecipitation studies with that antibody
demonstrated that the 7 subunit is associated with a SRC kinase.
Antagonists of tyrosine phosphorylation inhibited the acetylcholine-initiated
acrosome reaction, suggesting the involvement of a SRC kinase in the acrosome
reaction.
Received for publication, March 4, 2005 , and in revised form, April 18, 2005.
* This work was supported by National Institutes of Health Grant HD 33368 (to S. M.) and a postdoctoral fellowship (to P. K.) from National Institutes of Health Fertilization and Early Development Training Grant 5 T32 HD007131. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
To whom correspondence should be addressed: Dept. of Cell Biology and Human
Anatomy, University of California School of Medicine, Davis, CA 95616. Tel.:
530-752-3213; Fax: 530-752-8520; E-mail:
pkumar{at}ucdavis.edu.
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