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Mol. Hum. Reprod. Advance Access published online on May 7, 2008
Molecular Human Reproduction, doi:10.1093/molehr/gan026
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© The Author 2008. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org

Role of Cathepsins in Blastocyst Hatching in the Golden Hamster

G.V. Sireesha1, R.W. Mason2, M. Hassanein2, S. Tonac3, A. Navarrete Santos3, B. Fischer3 and P.B. Seshagiri1,4

1Department of Molecular Reproduction, Development and Genetics, Indian Institute of Science, Bangalore, India. 560012. 2Department of Biomedical Research, Alfred I. DuPont Hospital for Children, 1600 Rockland Road, Wilmington, DE 19803, U.S.A. 3Department of Anatomy and Cell Biology, Martin Luther University, D-06097, Halle (Saale), Germany.

4 To whom correspondence should be addressed Prof. Polani B. Seshagiri Department of Molecular Reproduction, Development and Genetics Indian Institute of Science Bangalore-560012. INDIA Phone: 91-080-22932687 Fax: 91-080-23600999 E-mail: polani{at}mrdg.iisc.ernet.in

The mammalian embryo is encased in a glycoproteinaceous coat, the zona pellucida (ZP) during preimplantation development. Prior to implantation, the blastocyst must undergo "hatching" or ZP escape. In hamsters, there is a thinning of the ZP followed by a focal lysis and a complete dissolution of the ZP during blastocyst hatching. Earlier studies from our laboratory have indicated a role for cysteine proteases in the hatching phenomenon. In this study, we tested the effect of specific inhibitors of the three classes of cysteine protease on blastocyst hatching. Cystatin, an endogenous cathepsin inhibitor, blocked blastocyst hatching. Similarly, Fmoc-Tyr-Ala-diazomethane, a synthetic cathepsin inhibitor, blocked hatching. Both showed dose-dependent and temporal inhibition of hatching. However, Z-Val-Ala-Asp-fluoromethylketone, a synthetic caspase inhibitor, and calpastatin, an endogenous calpain inhibitor, had no effect on hatching. The cathepsins were localized to blastocyst cells. Exogenous addition of cathepsins-L, -P or -B to cultured 8-cell embryos caused a complete ZP dissolution. The expression of mRNA and protein of cathepsins L and P was observed in peri-hatching blastocysts. Cathepsins -L and -P were detected in trophectodermal projections and in the ZP of peri-hatching blastocysts. These data provide the first evidence that blastocyst-derived cathepsins are functionally involved as zonalytic factors in the hatching of blastocysts in the golden hamster.

Key Words: blastocyst/cathepsins/hatching/hamster

Submitted on January 25, 2008; resubmitted on April 3, 2008; accepted on May 2, 2008.


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