HB-EGF directs stromal cell polyploidy and decidualization via cyclin D3 during implantation

Abstract

Stromal cell polyploidy is a unique phenomenon that occurs during uterine decidualization following embryo implantation, although the developmental mechanism still remains elusive. The general consensus is that the aberrant expression and altered functional activity of cell cycle regulatory molecules at two particular checkpoints G1 to S and G2 to M in the cell cycle play an important role in the development of cellular polyploidy. Despite the compelling evidence of intrinsic cell cycle alteration, it has been implicated that the development of cellular polyploidy may be controlled by specific actions of extracellular growth regulators. Here we show a novel role for heparin-binding EGF-like growth factor (HB-EGF) in the developmental process of stromal cell polyploidy in mice. HB-EGF, which is one of the earliest known molecular mediators of implantation in mice and humans, promotes stromal cell polyploidy via upregulation of cyclin D3. Adenoviral delivery of antisense cyclin D3 attenuates cyclin D3 expression and abrogates HB-EGF-induced stromal cell polyploidy in vitro and in vivo. Collectively, the results demonstrate that the regulation of stromal cell polyploidy and decidualization induced by HB-EGF depend on cyclin D3 induction.

Involvement of atypical transcription factor E2F8 in the polyploidization during mouse and human decidualization

“…Since HB-EGF has been shown to be regulated by progesterone during decidualization and tightly related to the polyploidization of decidual cells, 25,26 we assumed that HB-EGF may be involved in mediating the regulation of E2F8 expression in mouse uterus during decidualization. Compared with control group, the stromal cells treated with HB-EGF showed an increased proportion of tetraploid (Fig.…”

Conditional gene recombination by adenovirus-driven Cre in the mouse uterus

“…Previous studies in rodents have shown that the route of administration is a major determinant for the transfection efficiency in somatic tissues by adenoviral recombinants (Huard et al, 1995). However, limited information is currently available for in vivo adenoviral gene delivery in the uterus (Tan et al, 2004). To explore the potential route for ADV-Cre delivery into uterine cells during early pregnancy, we first examined whether a direct infusion of the virus into the uterine lumen is suitable for conditionally targeting uterine cells.…”

“…Generation of the replication-defective adenoviral vector for bacteriophage P1 Cre gene (ADV-Cre) followed the technique described by us (Tan et al, 2004). In brief, a 1.2-kb fragment (Xba1/SmaI) of a Cre gene containing the N-terminal nuclear localization signal (NLS) was derived from an expression clone, pBS/RIP-Cre-GH (generously provided by Holger Kulessa and originally developed by Christopher Wright, Vanderbilt University) and subsequently subcloned into XbaI/ECoRV sites of the shuttle vector, pAdTrack-CMV.…”

“…The resultant plasmid was linearized with PmeI and subsequently cotransfected with pAdEasy-1 into E. coli BJ5183 for the selection of recombinant plasmid. The generation of the empty recombinant plasmid (Empty ADV) has previously been described by us (Tan et al, 2004). The viral packaging of these vectors was carried out by transfecting 293 cells (Tan et al, 2004).…”

“…The generation of the empty recombinant plasmid (Empty ADV) has previously been described by us (Tan et al, 2004). The viral packaging of these vectors was carried out by transfecting 293 cells (Tan et al, 2004). Viral particles were purified through CsCl density gradient centrifugation and stored at -70°C.…”