Regulation of Estrogen Sulfotransferase Expression in Leydig Cells by Cyclic Adenosine 3′,5′-Monophosphate and Androgen¹

Abstract

Estrogen sulfotransferase (EST) catalyzes the specific sulfonation and inactivation of estrogens. A common site for EST expression in mammalian species is the testicular Leydig cells. In previous in vivo studies, we have shown that testicular expression of EST is under the regulation of LH. Thus, EST expression in mouse Leydig cells was abolished by hypophysectomy, but could be restored by hCG injection. In this study, we have evaluated the downstream mechanisms by which LH exerts its regulatory effect on EST. Primary mouse Leydig cells were isolated and purified by collagenase digestion and Percoll density gradient centrifugation. They were cultured in serum-free medium at 32 C and treated with various agents for 24 or 48 h, and levels of EST messenger RNA and enzyme activity were determined. Consistent with the in vivo data suggesting an essential role of LH in regulating EST expression, treatment of primary mouse Leydig cells in vitro with 100 microM 8-bromo-dibutyryl cAMP [(Bu)2cAMP] increased EST expression 3- to 5-fold. The effect of (Bu)2cAMP was attenuated by the steroidogenesis inhibitor aminoglutethimide and was mimicked by the potent androgen 5alpha-dihydrotestosterone (5-DHT). The activity of 5-DHT in stimulating EST expression was blocked by the androgen receptor antagonist, hydroxyflutamide. These data suggested the involvement of androgen in (Bu)2cAMP-induced EST expression. Further evidence came from the study with interleukin-1beta, another agent known to suppress Leydig cell steroidogenesis by down-regulating P450c17 gene expression. Treatment of Leydig cells with 0.2 ng/ml interleukin-1beta inhibited (Bu)2cAMP-induced EST expression, which was overcome by the addition of 5-DHT. Finally, in the testis-feminized mouse (Tfm) in which the androgen receptor is nonfunctional due to a frameshift mutation, testicular EST expression is completely absent, whereas messenger RNAs of steroidogenic enzymes such as P450c17 and 3beta-hydroxysteroid dehydrogenase are relatively abundant. We conclude that, by acting as an autocrine or paracrine factor, androgen plays an essential role in the regulation of estrogen sulfotransferase expression in Leydig cell by LH and cAMP.

Glutathione Stimulates Sulfated Estrogen Transport by Multidrug Resistance Protein 1

“…Thus, it has been assumed that an export pump is involved in its efflux from the cell. Testicular expression of estrogen sulfotransferase is mainly localized in Leydig cells and is regulated by luteinizing hormone via modulation of cAMP levels (23,24). Interestingly, the testes of 9 -12-month-old est Ϫ/Ϫ mice displayed Leydig cell hypertrophy/hyperplasia, as well as seminiferous tubule disruption.…”

Adult Exposure to Di-N-Butyl Phthalate (DBP) Induces Persistent Effects on Testicular Cell Markers and Testosterone Biosynthesis in Mice

“…The third affected testicular key protein, SULT1E1, is an enzyme central for estrogen regulation and homeostasis in the testis. It catalyzes the sulfate conjugation of estrone and estradiol, leading to their inactivation and inability to bind to the estrogen receptors in the testis [ 39 , 40 ]. Expression of SULT1E1 appears to be a unique feature of the adult type of Leydig cells and is under control of LH [ 41 , 42 ].…”

A Nuclear Receptor-Mediated Xenobiotic Response and Its Implication in Drug Metabolism and Host Protection

“…Accordingly, initial studies on the regulation of SULTs have focused on steroid hormone receptors [99][100][101][102]. Interestingly, several SULT isoforms are known to be differentially expressed between males and females (sex dimorphism).…”