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Estrogen Sulfotransferase Is Highly Expressed Along the Bovine Epididymis and Is Secreted Into the Intraluminal Environment

From the Centre de Recherche en Biologie de la Reproduction and Département d'Obstétrique-Gynécologie, Faculté de Médecine, Université Laval, Quebec City, Quebec, Canada.

Correspondence to: Dr Robert Sullivan, Unité d'Ontogénie-Reproduction, Centre de Recherche du Centre Hospitalier de l'Université Laval (CHUQ), 2705 boulevard Laurier, T1-49, Quebec City, Quebec, Canada G1V 4G2 (e-mail: robert.sullivan{at}crchul.ulaval.ca).

Abstract

Estrogen is found in high concentrations in the excurrent duct, where it regulates the expression of genes involved in water reabsorption. Estrogen sulfotransferase (EST) is a cytosolic enzyme that catalyzes specific sulfonation with a high affinity for estrogens. Because sulfated estrogens do not bind to estrogen receptors, they are considered to be hormonally inactive. EST may thus determine where along the male tract estrogenic environment predominates. Sulfotransferase activity increases along the epididymis and may also play a role in sperm physiology during the epididymal transit. Using a bovine model, we investigated the distribution of EST along the excurrent duct and the possibility that sterols associated with spermatozoa can be substrates of this enzyme. Reverse transcription polymerase chain reactions showed that mRNA encoding EST was expressed in the testis and all along the epididymis. A highly specific antiserum was raised against the bovine recombinant EST and used in Western blots and immunohistologic studies. Western blots of tissue homogenates showed that EST was localized all along the excurrent duct with a higher signal in the caput and corpus epididymidis. EST was detectable in the intraluminal compartment only in the caput epididymidis, where it was associated with epididymosomes and spermatozoa. EST was undetectable in different fractions of fluids collected in the cauda segment. In immunohistologic studies, EST was restricted to the acrosomal region of the caput, but not the cauda epididymal spermatozoa, and detectable in the cytoplasm of the epithelium bordering the lumen all along the epididymis as well as in the rete testis and vas efferens. This enzyme was also associated with the nucleus in the caput and corpus as well as with the apical membrane of the corpus epididymal epithelium. When recombinant EST was incubated in vitro in the presence of caput and cauda spermatozoa, it was able to add sulfate to sperm membrane cholesterol. Our study shows that EST is present in both the intracellular and intraluminal compartments of the epididymis, suggesting that this enzyme plays different roles along the excurrent duct.

     Key words: Reproductive tract, sperm maturation, cholesterol, cholesteryl sulfate

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