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J Biol Chem, Vol. 273, Issue 17, 10470-10474, April 24, 1998

Acrosin Accelerates the Dispersal of Sperm Acrosomal Proteins during Acrosome Reaction

Kazuo Yamagata, Keitaro Murayama, Masaru Okabe, Kiyotaka Toshimori^§, Tomoko Nakanishi, Shin-ichi Kashiwabara, and Tadashi Baba

From the Institute of Applied Biochemistry and Tsukuba Advanced Research Alliance, University of Tsukuba, and the National Institute for Advanced Interdisciplinary Research, Tsukuba Science City, Ibaraki 305-8572, the  Research Institute for Microbial Diseases, Osaka University, Yamadaoka 3-1, Suita, Osaka 565-0871, and the ^§ Department of Anatomy, Miyazaki Medical College, Miyazaki 889-1692, Japan

Using homologous recombination, we have previously produced male mice carrying a disruptive mutation (Acr^/) in the acrosin gene. Although Acr^/ mouse sperm lacking the acrosin protease activity still penetrated the zona pellucida and fertilized the egg, the mutant sperm exhibited a delay in penetration of the zona pellucida solely at the early stages after insemination. To further elucidate the role of acrosin in fertilization, we have examined the involvement of acrosin in the acrosome reaction of sperm using the Acr^/ mutant mice. When the ability of sperm to adhere (attach) and bind to the zona pellucida of cumulus-free eggs was assessed in vitro, no significant difference was observed among Acr^+/+, Acr^+/, and Acr^/ mouse sperm. Immunocytochemical analysis demonstrated that the release of several acrosomal proteins from the acrosome of Acr^/ mouse sperm was significantly delayed during the calcium ionophore- and solubilized zona pellucida-induced acrosome reaction, despite normal membrane vesiculation. These data indicate that the delayed sperm penetration of the zona pellucida in the Acr^/ mouse results from the altered rate of protein dispersal from the acrosome and provide the first evidence that the major role of acrosin is to accelerate the dispersal of acrosomal components during acrosome reaction.

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