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BOR - Papers in Press, published online ahead of print July 18, 2007.
Biol Reprod 2007, 10.1095/biolreprod.107.060004
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BIOLOGY OF REPRODUCTION 77, 794–802 (2007)
DOI: 10.1095/biolreprod.107.060004
© 2007 by the Society for the Study of Reproduction, Inc.

Dipeptidase-Inactivated tACE Action In Vivo: Selective Inhibition of Sperm-Zona Pellucida Binding in the Mouse1

Eishi Deguchi 3 4, Taeko Tani 3 4 5, Hitomi Watanabe 4, Shuichi Yamada 6, and Gen Kondoh 2 4 5

Bioscience Education and Research Center,6 Akita University, Akita City, Akita 010-8543, Japan Laboratory of Animal Experiments for Regeneration,4 Institute for Frontier Medical Sciences, Kyoto University, Sakyo-ku, Kyoto 606-8507, Japan CREST Programme,5 Japan Science and Technology Society, Sakyo-ku, Kyoto 606-8507, Japan

ABSTRACT

The angiotensin-converting enzyme (ACE) plays a crucial role in male fertilization and is a key regulator of blood pressure. Testicular ACE (tACE), the germinal specific isozyme expressed on different promoters, exclusively carries out the role of ACE in fertility, although the site and mode of action are not well known. To investigate the contribution of tACE in fertilization, we produced transgenic mouse lines carrying a dipeptidase-inactivated mutant. Although the transgenic mice showed normal blood pressure, kidney morphology, and fertility, reduced fertilization was observed after in vitro fertilization (IVF). The sperm-zona pellucida (ZP) binding was exclusively impaired in these lines in a manner similar to that observed in an Ace knockout mouse. The dipeptidase activity was reduced in epididymal ingredients but not in the testis. Furthermore, direct application of mutant protein did not suppress sperm-ZP binding of intact sperm during IVF, implying that the dipeptidase-inactivated mutant affects sperm modification in the epididymis for ZP binding. Our results indicate that the dipeptidase-inactivated tACE acts in vivo, suggesting that tACE contributes to fertilization as a dipeptidase at least in the epididymis.

angiotensin-converting enzyme, dipeptidase, epididymis, fertilization, in vitro fertilization, sperm, testis, transgenic mouse

FOOTNOTES

1Supported by grants from the Uehara Memorial Foundation and the Ministry of Education, Science, Sports, and Culture of Japan.

3These authors contributed equally to this work.

Correspondence: 2Gen Kondoh, Laboratory of Animal Experiments for Regeneration, Institute for Frontier Medical Sciences, Kyoto University, 53 Syogoin-kawahara-cho, Sakyo-ku, Kyoto 606-8507, Japan. FAX: 81 75 751 4860; e-mail: kondohg{at}frontier.kyoto-u.ac.jp






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Copyright © 2007 by the Society for the Study of Reproduction.