Am J Physiol Cell Physiol AJP: Endocrinology and Metabolism
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Am J Physiol Cell Physiol 274: C1496-C1500, 1998;
0363-6143/98 $5.00
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Vol. 274, Issue 6, C1496-C1500, June 1998

SNAP-25 is essential for cortical granule exocytosis in mouse eggs

Yoshihide Ikebuchi, Nobuyuki Masumoto, Tetsu Matsuoka, Takeshi Yokoi, Masahiro Tahara, Keiichi Tasaka, Akira Miyake, and Yuji Murata

Department of Obstetrics and Gynecology, Osaka University Medical School, Osaka 565, Japan

Synaptosome-associated protein of 25 kDa (SNAP-25) has been shown to play an important role in Ca2+-dependent exocytosis in neurons and endocrine cells. During fertilization, sperm-egg fusion induces cytosolic Ca2+ mobilization and subsequently Ca2+-dependent cortical granule (CG) exocytosis in eggs. However, it is not yet clear whether SNAP-25 is involved in this process. In this study, we determined the expression and function of SNAP-25 in mouse eggs. mRNA and SNAP-25 were detected in metaphase II (MII) mouse eggs by RT-PCR and immunoblot analysis, respectively. Next, to determine the function of SNAP-25, we evaluated the change in CG exocytosis with a membrane dye, tetramethylammonium-1,6-diphenyl-1,3,5-hexatriene, after microinjection of a botulinum neurotoxin A (BoNT/A), which selectively cleaves SNAP-25 in MII eggs. Sperm-induced CG exocytosis was significantly inhibited in the BoNT/A-treated eggs. The inhibition was attenuated by coinjection of SNAP-25. These results suggest that SNAP-25 may be involved in Ca2+-dependent CG exocytosis during fertilization in mouse eggs.

fertilization; soluble N-ethylmaleimide-sensitive factor attachment protein receptor; botulinum neurotoxin A


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