Localization of connective tissue growth factor in human uterine tissues

Abstract

Connective tissue growth factor (CTGF) is a recently described heparin-binding mitogen for fibroblasts and smooth muscle cells. The aim of this study was to investigate the production of CTGF by human uterine tissues using immunohistochemical and Northern blotting analyses. For immunohistochemistry, formalin-fixed human proliferative (n = 5), early secretory (n = 5; days 15-19), mid-secretory (n = 5; days 20-23), late secretory (n = 5; days 24-28) endometrial, and decidual (n = 5) tissues were stained using a highly specific affinity-purified polyclonal antibody raised against residues 81-94 of human CTGF. Myometrial (n = 5) and leiomyoma (n = 5) tissues were also used for CTGF immunochemistry. In proliferative endometrium, epithelial and vascular endothelial cells showed strong CTGF immunoreactivity, whereas stromal cells were negative or only weakly positive for the CTGF protein. Throughout the entire secretory stage, CTGF was detected in epithelial and endothelial cells of endometrium. Stromal cells showed strong immunoreactivity to CTGF only in oedematous areas for early and mid-secretory endometrium, and in decidualized regions of late secretory endometrium. During pregnancy, the decidual, epithelial and endothelial cells of the endometrium were all immunoreactive to CTGF. In myometrial and leiomyoma samples, CTGF immunoreactivity was found only in the endothelial cells. Northern blotting of mRNA from normal uterus (n = 2) or leiomyoma (n = 6) using a 320 bp human CTGF cDNA probe revealed a single 2.4 kb transcript. This study is the first to demonstrate CTGF gene expression and localization of its encoded protein in human uterine tissues. The cell- and cycle-specific localization of CTGF support a role for this molecule in regulating aspects of uterine cell growth, migration, and/or matrix production during the menstrual cycle and pregnancy.

Spatial differences in gene expression in the bovine oviduct

“…In the uterus of mice and humans, during the luteal phase, CTGF is expressed in luminal and glandular epithelium (Surveyor et al 1998, Uzumcu et al 2000. During early pregnancy, this gene is upregulated in the uterus of mice from Day 1.5 to 3.5 compared with Day 4.5 (Surveyor et al 1998) and in the endometrium of pregnant cattle compared with non-pregnant cattle at Day 13, 16 and 18 post oestrus (Klein et al 2006, Forde et al 2010.…”

Early postnatal methoxychlor exposure inhibits folliculogenesis and stimulates anti-Mullerian hormone production in the rat ovary

“…One of the sections was used for IHC as previously described (Uzumcu et al 2000). The sections were deparaffinized, rehydrated, and microwaved in 0 .…”

The heparin-binding 10 kDa fragment of connective tissue growth factor (CTGF) containing module 4 alone stimulates cell adhesion

“…Ovarian CTGF appears to be involved in recruitment and mitosis of theca cells and maintenance of the corpus luteum (Wandji et al 2000, Slee et al 2001, Liu et al 2002 while uterine CTGF has been implicated in angiogenesis and extracellular matrix remodeling during placentation and decidualization. , Moussad & Brigstock 2000, Uzumcu et al 2000, Moussad et al 2002. Expression of CTGF in the ovary is regulated by TGF-and gonadotrophins , Liu et al 2002 while in the uterus it is regulated by TGF-, estrogen and progesterone (Rageh et al 2001).…”

“…Estrogen and progesterone also regulate the production of cysteine-rich 61 (CYR61) which is structurally related to CTGF (Sampath, Zhu et al 2001, Xie, Miller et al 2001, Sampath et al 2002. Both CYR61 and CTGF are expressed in steroid-dependent tumors of the breast or uterus (Frazier & Grotendorst 1997, Tsai et al 2000, Uzumcu et al 2000, Sampath, Zhu et al 2001, Xie, Miller et al 2001, Sampath et al 2002, Tsai et al 2002.…”