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Institute for Hormone and Fertility Research, University of Hamburg (R.T., B.G.), Hamburg; and the Institute of Anatomy, University of Frankfurt (E.M.), Germany; and the Institute of Medical Biochemistry, University of Oslo (K.T.), Oslo, Norway
Address all correspondence and requests for reprints to: Dr. Birgit Gellersen, Institute for Hormone and Fertility Research, Division of Reproductive Sciences, University of Hamburg, Grandweg 64, 22529 Hamburg, Germany. E-mail: 100607.1557{at}compuserve.com
Decidualization of human endometrial stromal (ES) cells in
vitro is induced by cAMP analogs and ligands that elevate
cellular cAMP levels. A marker of this differentiation process is the
activation of the decidual PRL (dPRL) promoter. In a primary ES cell
culture system we show that relaxin not only acutely but permanently
elevates cellular cAMP levels and leads to induction of PRL secretion
after 6 days. Northern and Western blot analyses revealed that all
regulatory subunit isoforms (RI
, RIß, RII, and RIIß) and
catalytic subunits C and Cß of protein kinase A (PKA) are
expressed in ES cells. Transcript levels of PKA subunit isoforms are
not altered during decidualization, but in decidualized ES cells,
exposed to relaxin for more than 6 days, a significant reduction of
RI protein level occurs, whereas levels of all other forms remain
unchanged. Reduction of R subunits might result in a net increase in
free C subunit activity. This alteration is not due to a change in the
mitotic state of the cells, as proliferating cell nuclear antigen is
evenly expressed in undifferentiated and differentiated ES cell
cultures. In transient transfections of undifferentiated ES cells, the
dPRL promoter is activated by 8-bromo-cAMP and the C subunit (Cß) of
PKA. This induction as well as the differentiation-dependent activity
of the dPRL promoter in transfected decidualized cells are effectively
abolished by the coexpression of protein kinase inhibitor. We
demonstrate that 332 bp of the dPRL promoter are sufficient to mediate
full inducibility by cAMP. Activation of the dPRL promoter by cAMP in
ES cells occurs in two steps: an initial weak induction within 12
h and a subsequent, much more pronounced induction after 12 h. The
secondary induction is not seen with a control construct driven by a
consensus cAMP response element (CRE) linked to a minimal promoter and
is absent from a uterine cell line that does not express the endogenous
dPRL gene. The early response of the dPRL promoter depends upon a
noncanonical CRE at position -12, as mutation of this sequence leads
to abolition of the early, but not the delayed, induction. The major
activation depends upon a different region within 332 bp of the dPRL
promoter; is probably indirect, as it follows different kinetics
compared to a classical CRE-mediated response; and is specific to ES
cells.
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K. J. Greenland, I. Jantke, S. Jenatschke, K. E. Bracken, C. Vinson, and B. Gellersen The Human NAD+-Dependent 15-Hydroxyprostaglandin Dehydrogenase Gene Promoter Is Controlled by Ets and Activating Protein-1 Transcription Factors and Progesterone Endocrinology, February 1, 2000; 141(2): 581 - 597. [Abstract] [Full Text] [PDF]
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T. Maruyama, Y. Yoshimura, and H. Sabe Tyrosine Phosphorylation and Subcellular Localization of Focal Adhesion Proteins during in Vitro Decidualization of Human Endometrial Stromal Cells Endocrinology, December 1, 1999; 140(12): 5982 - 5990. [Abstract] [Full Text]
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J. J. Brosens, N. Hayashi, and J. O. White Progesterone Receptor Regulates Decidual Prolactin Expression in Differentiating Human Endometrial Stromal Cells Endocrinology, October 1, 1999; 140(10): 4809 - 4820. [Abstract] [Full Text]
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Y. Pohnke, R. Kempf, and B. Gellersen CCAAT/Enhancer-binding Proteins Are Mediators in the Protein Kinase A-dependent Activation of the Decidual Prolactin Promoter J. Biol. Chem., August 27, 1999; 274(35): 24808 - 24818. [Abstract] [Full Text] [PDF]
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A. Einspanier, R. Nubbemeyer, S. Schlote, M. Schumacher, R. Ivell, K. Fuhrmann, and A. Marten Relaxin in the Marmoset Monkey: Secretion Pattern in the Ovarian Cycle and Early Pregnancy Biol Reprod, August 1, 1999; 61(2): 512 - 520. [Abstract] [Full Text]
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J. J. Kim, R. C. Jaffe, and A. T. Fazleabas Insulin-Like Growth Factor Binding Protein-1 Expression in Baboon Endometrial Stromal Cells: Regulation by Filamentous Actin and Requirement for de Novo Protein Synthesis Endocrinology, February 1, 1999; 140(2): 997 - 1004. [Abstract] [Full Text]
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H. Ungefroren, B. Gellersen, N. B. Krull, and H. Kalthoff Biglycan Gene Expression in the Human Leiomyosarcoma Cell Line SK-UT-1. BASAL AND PROTEIN KINASE A-INDUCED TRANSCRIPTION INVOLVES BINDING OF Sp1-LIKE/Sp3 PROTEINS IN THE PROXIMAL PROMOTER REGION J. Biol. Chem., October 30, 1998; 273(44): 29230 - 29240. [Abstract] [Full Text] [PDF]
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J. J. Kim, R.C. Jaffe, and A.T. Fazleabas Comparative Studies on the In Vitro Decidualization Process in the Baboon (Papio anubis) and Human Biol Reprod, July 1, 1998; 59(1): 160 - 168. [Abstract] [Full Text]
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 G Keryer, E Alsat, K Tasken, and D Evain-Brion Cyclic AMP-dependent protein kinases and human trophoblast cell differentiation in vitro J. Cell Sci., January 4, 1998; 111(7): 995 - 1004. [Abstract] [PDF]
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