Control and expression of cystatin C by mouse decidual cultures

Abstract

During mouse embryo implantation, trophoblast invasion is controlled in part by a balance of trophoblast-derived proteinases and uterine decidual proteinase inhibitors. Our work has focused on cystatin C, the secreted inhibitor of cathepsins B and L. We have previously shown that cystatin C is synthesized by the uterine decidua and localized to the cells in close contact with the trophoblast during implantation in vivo. In the work reported here we have established that decidualizing cultures show a similar upregulation of cystatin C. Using Northern and Western blotting and immunolocalization techniques both cystatin C mRNA and secreted protein increased with the morphological differentiation of stromal or decidual capsule cultures. In an effort to understand the regulation of cystatin C expression, decidual cells were analyzed under various culture conditions. Cystatin C expression was upregulated by increased cell density and by the presence of serum in the media. The growth factors TGF-beta(1) and EGF were found to induce cystatin C to levels comparable to serum stimulation. Co-culture with ectoplacental cones (EPCs) likewise induced expression and resulted in the localization of cystatin at the decidua:trophoblast interface. This work shows that decidualizing cultures are a good system to study cystatin C expression and that the expression is controlled in part by TGF-beta(1) and EGF signaling.

Increased plasma levels of cystatin C and transforming growth factor-β1 in patients with coronary artery ectasia: can there be a potential interaction between cystatin C and transforming growth factor-β1

“…TGF-b1 has also been shown to play a complex role in the pathogenesis of atherosclerosis [16][17][18][19]. It has been previously demonstrated that TGF-b1 clearly regulates cys C expression in Levels of cystatin C in coronary artery ectasia Yetkin et al 213 murine embryo [20], uterine decidual [35]. TGF-b1 has been found to be a potent inducer of cys C in vascular smooth muscle cell [14].…”

Localization of chondromodulin-I at the feto-maternal interface and its inhibitory actions on trophoblast invasion in vitro

“…Isolation of decidual cells was carried out as described previously [36] with minor modifications. Briefly, decidual capsules were removed from ICR pregnant mice at 7.5 days p.c.…”

Localization of Cathepsins D and B at the Maternal-Fetal Interface and the Invasiveness of the Trophoblast during the Postimplantation Period in the Mouse

“…In reproductive tissues, the expression of cathepsins and their inhibitors at the maternalfetal interface during stromal invasion by the trophoblast has also been pointed out as a putative determinant factor in this process [Denker et al, 1978;Denker, 1980Denker, , 1982Li et al, 1991;Afonso et al, 1997Afonso et al, , 1999Afonso et al, , 2002Screen et al, 2008;Baston-Buest et al, 2010].…”

“…Ectoplacental cones and decidual cell cocultures induced expression and resulted in the localization of cystatin at the interface between maternal and fetal cells in a transforming growth factor-beta (TGF-␤ )1 and EGF 2011;193:417-425 423 partially controlled pattern [Afonso et al, 2002]. Imbalance between cathepsins and cystatins has been associated with control of trophoblast invasion, differentiation of the endometrial stroma [Jokimaa et al, 2001], recurrent miscarriage [Nakanishi et al, 2005], and various diseases, especially cancer [Ebert et al, 1994;Henskens et al, 1996].…”

“…Afonso et al [1997Afonso et al [ , 1999Afonso et al [ , 2002 showed that cathepsins B and L are present in the conditioned medium of cultured trophoblast cells while cystatin C, a cathepsin inhibitor, is localized at the decidual compartment, suggesting a control for trophoblast invasiveness.…”