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Regular Article |
a Department of Cell Biology, Faculty of Pharmacy, University of Calabria, 87030 Rende (CS), Italy
b Department of Biochemistry, University of Caen, UPRES EA 2608, 14032-Caen, France
c Green Center for Reproductive Biology and the Department of Pharmacology, University of Texas Southwestern Medical Center, Dallas, Texas 75390-8857
d Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas, Texas 75390-8857
ABSTRACT
It has long been recognized that individual cell types within the testes possess the capacity to synthesize estrogen. A number of studies on different species have demonstrated that the levels of aromatase expression and the patterns of regulation are distinct between the different cell types of the testes. Whereas a variety of promoters have been shown to contribute to the patterns of aromatase expression in different cell lineages, studies using ovarian RNA, testis RNA, and Leydig cell tumor lines have demonstrated that the same promoter (promoter II) was used in each. Recent experiments using potent aromatase inhibitors or analysis of animals in which the genes encoding the estrogen receptor-alpha (ER-
) or the aromatase, P450, are defective, have confirmed the importance of local estrogen formation in normal testicular function. In order to permit experiments to identify the elements controlling aromatase expression in the individual cell compartments of the testes, we prepared RNA from purified preparations of Leydig, Sertoli, and germ cells. Using specific oligonucleotide primers, the sites of initiation of the aromatase mRNA were determined using rapid amplification of cDNA ends (RACE) and nucleotide sequence analysis of the resulting cDNA fragments. Our results indicate that aromatase mRNA is derived from the proximal promoter (PII) of the aromatase gene in each of the major cell types of the rat testes.
First decision: 25 October 2000.
1 Supported by grant I-1090 from the Robert A. Welch Foundation, and DK03892 from the National Institutes of Health.
2 Correspondence: Michael J. McPhaul, Department of Internal Medicine, UT Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas, TX 75390-8857. FAX: 214 648 8917; michael.mcphaul{at}utsouthwestern.edu
3 These authors contributed equally to this work.
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