Comparison of the effect of oestradiol, tamoxifen and raloxifene on nerve growth factor-alpha expression in specific neonatal mouse uterine cell types using laser capture microdissection

Abstract

Oral dosing of CD-1 mice on days 2-5 after birth with tamoxifen but not raloxifene disrupts the development of the myometrium, resulting in adult uterine adenomyosis. Using laser capture microdissection and RT-PCR we have investigated nerve growth factor (NGF) and cognate receptor expression in uterine cells of 6-day-old pups that may be important in early developmental changes that give rise to adenomyosis. NGF down-regulation is known to occur during terminal myogenic differentiation.NGF was found exclusively in endometrial luminal epithelium of controls. It was up-regulated 18-fold in the luminal epithelium following dosing with tamoxifen but not raloxifene. Western blotting for NGF protein in the whole uterus showed a 25-fold increase after tamoxifen treatment. Expression of the low affinity p75 neutrophin receptor (p75 NTR ) was twofold higher in the myometrium compared with luminal epithelium or stroma. This was not altered following tamoxifen treatment. There was no detectable expression of high affinity tyrosine kinase receptor (trkA NGFR ). This study shows luminal epithelial cells of the endometrium primarily form NGF. This suggests that NGF normally regulates the differentiation of the mesenchyme into uterine myocytes through paracrine mechanisms and that an early disturbance of this process plays a key role in the subsequent development of adenomyosis.

The effects of tamoxifen and estradiol on myometrial differentiation and organization during early uterine development in the CD1 mouse

“…Tamoxifen, but not estradiol neonatal exposure, resulted in the development of adenomyosis in CD1 mice as early as day 10, with the disruption of the inner myometrial layer and ingrowth of endometrial glands and stroma. The doses of tamoxifen and estradiol used in our work were derived from the dose-response studies , and have been previously described and utilized in the similar experiments , Green et al 2003.…”

Analysis of estrogen-regulated genes in mouse uterus using cDNA microarray and laser capture microdissection

“…LCM allows the isolation of specific uterine cell types without contamination from other cell types. Green et al (2003) showed that mRNA from cells could be successfully isolated by LCM from frozen sections with high efficiency, producing RNA with excellent quality and quantity. We isolated populations of specific uterine cell types from frozen sections stained with Mayer's hematoxylin: luminal epithelial, stromal and muscle cells.…”

“…We also employed a combination of laser capture microdissection (LCM) and RT-PCR to quantify estrogen-responsive genes in specific cell types of the ovariectomized mouse uterus. Recently, LCM has been used to isolate specific cell types from tissue sections for the purpose of differential gene expression (Green et al 2003). Using these techniques, we profiled both early and late responsive genes regulated by estrogen and demonstrated that several responsive genes were differentially expressed in specific cell types of the estrogen-induced uterus.…”

Estrogen in combination with 5-azacitidine up-regulates p75NTR expression and induces apoptosis in 22Rv1 prostate cancer cells

“…A number of studies suggest that estrogen can alter the expression of neurotrophin receptors and perhaps even neurotrophins (16,17,35). We first demonstrated that estrogen enhances the expression of p75NTR in 22Rv1 prostate cancer cells, This effect occurs in association with the up-regulated expression of ESR2, suggesting that estrogen might not only directly regulate p75NTR expression, but may also influence the neurotrophin pathways via their receptors.…”

“…Estrogen receptors are co-localized in cells that express neurotrophins and their receptors, and estrogen further regulates the expression of the neurotrophic system in many tissues, not just the nervous system (14,15). Estrogen treatment can induce the expression of p75NTR in Estrogen in combination with 5-azacitidine up-regulates p75NTR expression and induces apoptosis in 22Rv1 prostate cancer cells uterine cells and in cholinergic neurons (16,17), but little is known regarding its effects in the prostate. We speculated that interactions between estrogen and the neurotrophic system may partially explain the beneficial effects of estrogen therapy for androgen-refractory prostate cancer.…”