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This version published online on November 4, 2004
Endocrinology, doi:10.1210/en.2004-1224
A more recent version of this article appeared on February 1, 2005
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Submitted on September 15, 2004
Accepted on October 25, 2004

Interferon Stimulated Gene 15 (ISG15) Conjugates to Endometrial Cytosolic Proteins and is Expressed at the Uterine-Placental Interface Throughout Pregnancy in Sheep*

Margaret M. Joyce, Frankie J. White, Robert C. Burghardt, Jesus J. Muñiz, Thomas E. Spencer, Fuller W. Bazer, and Greg A. Johnson*

Center for Animal Biotechnology and Genomics, Department of Veterinary Integrative Biosciences, College of Veterinary Medicine and Biomedical Sciences, and Department of Animal Science, College of Agriculture and Life Sciences, Texas A&M University, College Station, TX 77843-4458

* To whom correspondence should be addressed. E-mail: gjohnson{at}cvm.tamu.edu.

Interferon stimulated gene 15 (ISG15) is a ubiquitin homolog expressed in uteri of ruminants in response to interferon-{tau} (IFN{tau}), and is also induced during pregnancy in the uteri of mice, pigs, humans, and baboons. This study examined expression of ISG15 and its conjugation to target proteins in the ovine uterus beyond the period of IFN{tau}. secretion by the conceptus. Although steady-state levels of ISG15 mRNA decreased after Day 25 of pregnancy, ISG15 persisted in endometrium through Day 120. In situ hybridization and immunocytochemistry localized ISG15 across the entire uterine wall through Day 25, after which expression was restricted to endometrial stroma along the maternal-placental interface. Western blots revealed ISG15 and ISG15-conjugated proteins in endometrium. Treatment of ovariectomized sheep with progesterone and IFN{tau}. increased both free and conjugated ISG15. These results are the first to show in vivo regulation of ISG15 function (i.e. conjugation to target proteins) by a Type I IFN in the uterus of any species, and that ISG15 is expressed at contacts between the placenta and uterus when trophectoderm no longer produces IFN{tau} Interestingly, mRNA for the Type II IFN {gamma} was present in the endometrial stromal compartment on Days 15-50 which may stimulate the synthesis of ISG15 through later pregnancy. We hypothesize that ISG15 is not merely a consequence of an anti-viral state induced by trophoblast IFN{tau}, but represents a critical component of the microenvironment at the uterine-placental interface during the progressive events of conceptus development, implantation, and placentation in sheep, and perhaps other mammalian species.
Key words: ISG15 • IFN-tau • IFN-gamma • ovine • uterus • pregnancy




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