Matrix-Metalloproteinases-2 and -9 Production in Bovine Endometrial Cell Culture.

Abstract

Abstract. In vitro cell culture is a convenient tool for studying cellular mechanisms. In the present study, production of matrix-metalloproteinases (MMPs) in bovine endometrial (containing both epithelial and stromal cells) monolayer cells was examined. Blastocysts attached to the endometrial cells in a monolayer culture were examined for their effects on MMP-2 production. Initial attachment of blastocysts to the monolayer inhibited MMP-2 production by endometrial cells. But once trophoblast cells began to migrate into the endometrial cell layer, MMP-2 production increased, and at the same time MMP-9 production also became evident in the medium. In order to understand how blastocysts affected MMP-2 production, we examined the effect of progesterone, estradiol, insulin-like growth factors (IGFs), tumor necrosis factors (TNFs), and interferon-tau (IFN-τ) supplementation. It was IFN-τ that inhibited the production of MMP-2. In addition, progesterone at a lower dose appeared to inhibit MMP-2 production. Both TNF-α and TNF-β strongly stimulated the production of MMP-2 and MMP-9, whereas IGFs had no effect. Based on these findings, it appears that conceptus has the capacity to inhibit MMP activity. Key words: MMPs, Cytokine, Endometrium, Cell culture, Implantation (J. Reprod. Dev. 49: [45][46][47][48][49][50][51][52][53] 2003) any factors including proteases, growth factors, cytokines and steroid hormones play i m p or t an t r ol es i n t is s u e r em od e li n g a n d coordination of endometrial function, resulting in a suitable environment for embryo attachment during early pregnancy [1][2][3][4][5][6]. Asynchrony between the conceptus and endometrium causes failure of implantation, leading to early embryonic l o s s , s o th a t an at o m i c a l an d b i o c h e m i c a l modifications are essential for endometrial tissue remodeling and trophoblast development [7][8][9]. It is also w ell kn ow n th at r em odeli ng of the ext rac el lular m at rix (EC M) is es sen tial for implantation and placentation, as well as for inflammation, wound healing and tumorigenesis. The ECM is composed of various molecules including collagens, heparan sulfate proteoglycans and laminin and provides a suitable structural microenvironment for the embryo. It is necessary t h a t t h e E C M m u s t u n d e r g o p r o t e o l y t i c degradation during early pregnancy both in

Bovine endometrial metallopeptidases MMP14 and MMP2 and the metallopeptidase inhibitor TIMP2 participate in maternal preparation of pregnancy

“…Interferon-, produced in large amounts by the elongating embryo, has been shown to inhibit the expression of pro-MMP1 and pro-MMP2 as well as the activity of MMP2 (Hashizume et al, 2003), but in our study the expression of neither MMP14 nor MMP2 mRNA was reduced due to pregnancy. In sheep, interferon- has been shown to trigger the increase of TIMP2 (Chen et al, 2007;Gray et al, 2006), and TIMP2 mRNA expression has been shown to remain elevated in ovine and bovine early pregnancy (during Page 24 of 32 A c c e p t e d M a n u s c r i p t 17 implantation until day 31) (Ledgard et al, 2009).…”

A microarray analysis for genes regulated by interferon-τ in ovine luminal epithelial cells

“…IFNT down-regulated genes for several MMPs, up-regulated TIMP1, TIMP2, TIMP3, and TIMP4, and increased transcript concentrations for a number of genes encoding extracellular matrix components (Table 6). Others have demonstrated that IFNT inhibits production of MMPs in primary cultures of bovine endometrial cells (Salamonsen et al 1994, Hashizume et al 2003. In addition, these observations suggest that IFNT promotes extracellular matrix accretion rather than breakdown in the ovine uterus during the preimplantation period.…”

Analysis of Uteroplacental-specific Molecules and Their Functions During Implantation and Placentation in the Bovine

“…As in vitro cell culture system was used to elucidate the regulatory mechanisms underlying the role of MMPs in degrading the ECM in the bovine endometrium [50]. It was observed that MMP-2 production was insignificantly affected by steroid hormones; however, tumor necrosis factors (TNF-α and β) stimulated MMP-2 production.…”