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Regulated Expression of Inhibitor of Apoptosis Protein 3 in the Rat Corpus Luteum¹

School of Anatomy and Human Biology,³ The University of Western Australia, Crawley, Western Australia 6009, Australia, and the West Australian Institute of Medical Research, Sir Charles Gairdner Hospital, Shenton Park, Western Australia 6008, Australia Department of Clinical Research,⁴ Faculty of Medicine, University of Berne, Berne, Switzerland Department of Physiology,⁵ Morehouse School of Medicine, Atlanta, Georgia 30310-1495

We sought to investigate the role inhibitor of apoptosis proteins (IAPs) play in the life cycle of the corpus luteum (CL) of the rat. We isolated two clones with amino acid homology to rat IAP2 (BIRC 3) and three to rat IAP3 (rIAP3; BIRC 4). The expression of rIAP3 mRNA was examined in the rat CL during and after pregnancy, in Day 8 pregnant rats after 24-h treatment of gonadotropin-releasing hormone-agonist (GnRH-Ag), and in a CL organ culture model of spontaneous apoptosis in the absence of tropic support with and without superoxide dismutase. We used real-time RT-PCR to quantitate rIAP3 mRNA expression. Interestingly, a significant reduction in rIAP3 levels was seen at the time of CL regression in the course of natural pregnancy and the GnRH-Ag model. Surprisingly, rIAP3 mRNA levels in the CL organ culture model of spontaneous apoptosis failed to show significant changes, although TUNEL (terminal deoxynucleotide transferase-mediated dUTP nick end-labeling) reaction showed 30%–40% of the cells undergoing DNA fragmentation after 2 h in culture. In situ hybridization revealed that rIAP3 expression was localized to the cytoplasm of luteal and granulosa cells. These data clearly demonstrate both the presence of IAPs in the rat CL and the regulation of rIAP3 during in vivo apoptotic cell death, indicating a role for IAPs in the maintenance of CL function and demise.

¹ This study was supported by the following grants: the Australian Research Council, National Health and Medical Research Council, Raine Medical Foundation (all to A.M.D.), and the Dora Lush Biomedical Postgraduate Scholarship (to R.R.L.); the National Institutes of Health (GM08248) and by the National Aeronautics and Space Administration (NAG9-963), Washington, DC (to R.S.); and by grants from the Bernische Krebsliga, the Stiftung fur-klinisch-experimentelle Tumorforschung and the Swiss National Science Foundation (31-63700.00) (to R.R.F.).

² Correspondence: Arun M. Dharmarajan, School of Anatomy and Human Biology, University of Western Australia, 35 Stirling Highway, Crawley, WA 6009, Australia. FAX: 61 8 9380 1051; dharma{at}anhb.uwa.edu.au

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