α-SNAP and NSF are required in a priming step during the human sperm acrosome reaction

被引:52
作者
Tomes, CN
De Blas, GA
Michaut, MA
Farré, EV
Cherhitin, O
Visconti, PE
Mayorga, LS
机构
[1] Univ Nacl Cuyo, Fac Ciencias Med, Lab Biol Celular & Mol, Inst Histol & Embriol,IHEM,CONICET, RA-5500 Mendoza, Argentina
[2] Univ Virginia, Ctr Recombinant Gamete Contracept Vaccinogens, Dept Cell Biol, Charlottesville, VA 22903 USA
关键词
acrosome; exocytosis; NSF; alpha-SNAP; sperm;
D O I
10.1093/molehr/gah126
中图分类号
Q [生物科学];
学科分类号
07 ; 0710 ; 09 ;
摘要
The acrosome is a membrane-limited granule that overlies the nucleus of the mature spermatozoon. In response to physiological or pharmacological stimuli it undergoes a special type of Ca2+-dependent exocytosis termed the acrosome reaction (AR), which is an absolute prerequisite for fertilization. Aided by a streptolysin-O permeabilization protocol developed in our laboratory, we have previously demonstrated requirements for Rab3A, N-ethylmaleimide-sensitive factor (NSF), several soluble NSF-attachment protein receptor (SNARE) proteins, and synaptotagmin VI in the human sperm AR. Here, we show that alpha-soluble NSF-attachment protein (alpha-SNAP), a protein essential for most fusion events through its interaction with NSF and the SNARE complex, exhibits a direct role in the AR. First, the presence of alpha-SNAP is demonstrated by the Western blot of human sperm protein extracts. Immunostaining experiments reveal an acrosomal localization for this protein. Second, the Ca2+ and Rab3A-triggered ARs are inhibited by anti-alpha-SNAP antibodies. Third, bacterially expressed alpha-SNAP abolishes exocytosis in a fashion that depends on its interaction with NSF. Fourth, we show a requirement for alpha-SNAP/NSF in a prefusion step early in the exocytotic pathway, after the tethering of the acrosome to the plasma membrane and before the efflux of intra-acrosomal Ca2+. These results suggest a key role for alpha-SNAP/NSF in the AR, and strengthen our understanding of the molecular players involved in the vesicle-to-plasma membrane fusion taking place during exocytosis.
引用
收藏
页码:43 / 51
页数:9
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