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In Utero and In Vitro Proteinase Activity During the Mesocricetus auratus Embryo Zona Escape Time Window¹

^a Department of Biology, University of Southern Colorado, Pueblo, Colorado 81001 ^b Department of Animal Health & Biomedical Sciences, University of Wisconsin, Madison, Wisconsin 53711

ABSTRACT

The goal of the present study was to investigate proteinase activity in uterine flushates collected during the zona loss time window (68–80 h post-egg activation) in both pregnant and pseudopregnant hamsters and in culture medium conditioned by hatching blastocysts. Several prominent enzyme activities appeared in all pregnant and pseudopregnant uterine flushates. However, only a 45, 43 x 10^-3 M_r doublet coincided with the zona loss time window; these bands were absent outside of this time window and were not found in conditioned medium. In medium conditioned by hatching blastocysts, enzyme activity was represented by a 70, 65 x 10^-3 M_r doublet identical to a doublet seen in all uterine flushates collected and in serum. There were 12 pregnant and 8 pseudopregnant uterine flushates that were capable of zona lytic activity in vitro (positive bioassays). Of these positive bioassays, five pregnant and four pseudopregnant uterine flushates exhibited the 45, 43 x 10^-3 M_r doublet (correlative positive bioassays). These data suggest that there is an important uterine contribution to blastocyst escape from the zona pellucida, consisting of proteinases secreted during a finite time window prior to blastocyst attachment that are different from the proteinases responsible for the zona lytic activity in vitro.

FOOTNOTES

First decision: 28 March 2000.

¹ This study was funded by the National Institutes of Health Biomedical Research Support Grant Program S06 GM08197-17. Preliminary data from this study were presented at the 32nd annual meeting of the Society for the Study of Reproduction (Biol Reprod 1999; 60[suppl 1]:505).

² Correspondence: David Gonzales, Department of Biology, University of Southern Colorado, 2200 Bonforte Blvd., Pueblo, CO 81001. FAX: 719 549 2732; dgonz{at}uscolo.edu

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