INVESTIGADORES
RUBINSTEIN Marcelo
artículos
Título:
Impaired sperm fertilizing ability in mice lacking Cysteine-RIch Secretory
Autor/es:
VANINA G. DA ROS; JULIETA A. MALDERA; WILLIAM D. WILLIS; DÉBORA J. COHEN; EUGENIA H. GOULDING; DIEGO M. GELMAN; MARCELO RUBINSTEIN; EDWARD M. EDDY; PATRICIA S. CUASNICU
Revista:
DEVELOPMENTAL BIOLOGY
Editorial:
ACADEMIC PRESS INC ELSEVIER SCIENCE
Referencias:
Año: 2008 vol. 320 p. 12 - 18
ISSN:
0012-1606
Resumen:
Mammalian fertilization is a complex multi-step process mediated by different molecules present on both
gametes. Epididymal protein CRISP1, a member of the Cysteine-RIch Secretory Protein (CRISP) family, was
identified by our laboratory and postulated to participate in both spermzona pellucida (ZP) interaction and
gamete fusion by binding to egg-complementary sites. To elucidate the functional role of CRISP1 in vivo, we
disrupted the Crisp1 gene and evaluated the effect on animal fertility and several sperm parameters. Male
and female Crisp1−/− animals exhibited no differences in fertility compared to controls. Sperm motility and
the ability to undergo a spontaneous or progesterone-induced acrosome reaction were neither affected infied by our laboratory and postulated to participate in both spermzona pellucida (ZP) interaction and
gamete fusion by binding to egg-complementary sites. To elucidate the functional role of CRISP1 in vivo, we
disrupted the Crisp1 gene and evaluated the effect on animal fertility and several sperm parameters. Male
and female Crisp1−/− animals exhibited no differences in fertility compared to controls. Sperm motility and
the ability to undergo a spontaneous or progesterone-induced acrosome reaction were neither affected inin vivo, we
disrupted the Crisp1 gene and evaluated the effect on animal fertility and several sperm parameters. Male
and female Crisp1−/− animals exhibited no differences in fertility compared to controls. Sperm motility and
the ability to undergo a spontaneous or progesterone-induced acrosome reaction were neither affected inCrisp1 gene and evaluated the effect on animal fertility and several sperm parameters. Male
and female Crisp1−/− animals exhibited no differences in fertility compared to controls. Sperm motility and
the ability to undergo a spontaneous or progesterone-induced acrosome reaction were neither affected inCrisp1−/− animals exhibited no differences in fertility compared to controls. Sperm motility and
the ability to undergo a spontaneous or progesterone-induced acrosome reaction were neither affected in
Crisp1−/− mice. However, the level of protein tyrosine phosphorylation during capacitation was clearly lower
in mutant sperm than in controls. In vitro fertilization assays showed that Crisp1−/− sperm also exhibited a
significantly reduced ability to penetrate both ZP-intact and ZP-free eggs. Moreover, when ZP-free eggs were
simultaneously inseminated with Crisp1+/+ and Crisp1−/− sperm in a competition assay, the mutant sperm
exhibited a greater disadvantage in their fusion ability. Finally, the finding that the fusion ability of Crisp1−/−−/− mice. However, the level of protein tyrosine phosphorylation during capacitation was clearly lower
in mutant sperm than in controls. In vitro fertilization assays showed that Crisp1−/− sperm also exhibited a
significantly reduced ability to penetrate both ZP-intact and ZP-free eggs. Moreover, when ZP-free eggs were
simultaneously inseminated with Crisp1+/+ and Crisp1−/− sperm in a competition assay, the mutant sperm
exhibited a greater disadvantage in their fusion ability. Finally, the finding that the fusion ability of Crisp1−/−In vitro fertilization assays showed that Crisp1−/− sperm also exhibited a
significantly reduced ability to penetrate both ZP-intact and ZP-free eggs. Moreover, when ZP-free eggs were
simultaneously inseminated with Crisp1+/+ and Crisp1−/− sperm in a competition assay, the mutant sperm
exhibited a greater disadvantage in their fusion ability. Finally, the finding that the fusion ability of Crisp1−/−ficantly reduced ability to penetrate both ZP-intact and ZP-free eggs. Moreover, when ZP-free eggs were
simultaneously inseminated with Crisp1+/+ and Crisp1−/− sperm in a competition assay, the mutant sperm
exhibited a greater disadvantage in their fusion ability. Finally, the finding that the fusion ability of Crisp1−/−Crisp1+/+ and Crisp1−/− sperm in a competition assay, the mutant sperm
exhibited a greater disadvantage in their fusion ability. Finally, the finding that the fusion ability of Crisp1−/−finding that the fusion ability of Crisp1−/−
sperm was further inhibited by the presence of CRISP1 or CRISP2 during gamete co-incubation, supports that
another CRISP cooperates with CRISP1 during fertilization and might compensate for its lack in the mutant
mice. Together, these results indicate that CRISP proteins are players in the mammalian fertilization process.
To our knowledge this is the first knockout mice generated for a CRISP protein. The information obtained
might have important functional implications for other members of the widely distributed and evolutionarily
conserved CRISP family.first knockout mice generated for a CRISP protein. The information obtained
might have important functional implications for other members of the widely distributed and evolutionarily
conserved CRISP family.