Effects of the Antiestrogen Fulvestrant (ICI 182,780) on Gene Expression of the Rat Efferent Ductules1

Abstract

The efferent ductules express the highest amount of estrogen receptors ESR1 (ERalpha) and ESR2 (ERbeta) within the male reproductive tract. Treatment of rats with the antiestrogen fulvestrant (ICI 182,780) causes inhibition of fluid reabsorption in the efferent ductules, leading to seminiferous tubule atrophy and infertility. To provide a more comprehensive knowledge about the molecular targets for estrogen in the rat efferent ductules, we investigated the effects of ICI 182,780 treatment on gene expression using a microarray approach. Treatment with ICI 182,780 increased or reduced at least 2-fold the expression of 263 and 98 genes, respectively. Not surprisingly, several genes that encode ion channels and macromolecule transporters were affected. Interestingly, treatment with ICI 182,780 markedly altered the expression of genes related to extracellular matrix organization. Matrix metalloproteinase 7 (Mmp7), osteopontin (Spp1), and neuronal pentraxin 1 (Nptx1) were among the most altered genes in this category. Upregulation of Mmp7 and Spp1 and downregulation of Nptx1 were validated by Northern blot. Increase in Mmp7 expression was further confirmed by immunohistochemistry and probably accounted for the decrease in collagen content observed in the efferent ductules of ICI 182,780-treated animals. Downregulation of Nptx1 probably contributed to the extracellular matrix changes and decreased amyloid deposition in the efferent ductules of ICI 182,780-treated animals. Identification of new molecular targets for estrogen action may help elucidate the regulatory role of this hormone in the male reproductive tract.

Locally produced relaxin may affect testis and vas deferens function in rats

“…Rats were treated once a week for 2 months with corn oil (control group) or ICI 182,780 (10 mg/ rat, s.c., AstraZeneca), starting at an age of 30 days. This treatment does not alter plasma estradiol and testosterone (Yasuhara et al 2008).…”

“…Anti-estrogen treatment Animals were treated as previously described (Oliveira et al 2002, Yasuhara et al 2008. Rats were treated once a week for 2 months with corn oil (control group) or ICI 182,780 (10 mg/ rat, s.c., AstraZeneca), starting at an age of 30 days.…”

“…Sections of 5-7 mm thickness were cut and mounted on silanized slides (Dako, Carpinteria, CA, USA). Immunoperoxidase reaction was performed as previously described (Yasuhara et al 2008). Briefly, tissue sections were incubated for 5 min at room temperature with 0.1 M glycine in Tris-buffered saline (TBS, 20 mM Tris-HCl, 150 mM NaCl, pH 7.4).…”

Estrogen receptors and function in the male reproductive system

“…Using a microarray analysis of genes differentially expressed in the efferent ductules of fulvestrant-treated animals, we have been able to identify a significant impact of the antiestrogen treatment on gene expression, and besides genes related to ion and water transport, genes related to extracellular matrix organization and sperm maturation and/or capacitation, among others, were also affected, suggesting a broader role of estrogen to regulate the morphology and function of the efferent ductules (52).…”

From SELDI-TOF MS to protein identification by on-chip elution

“…In addition, a recently reported study suggested that LECT-1 may participate in the innate immune defense system of the disk abalone (Haliotis discus discus) [14]. Furthermore, it should be noted that a down-regulation of the LECT-1 gene expression has been observed in the rat male reproductive tract following treatment with the estrogen receptor antagonist Fulvestrant [15].…”