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Estrogen Receptor ß Expression and Apoptosis of Spermatocytes of Mice Overexpressing a Rat Androgen-Binding Protein Transgene¹

Grup de Recerca en Endocrinologia Molecular³ Departament d'Anatomia Patològica,⁴ Hospital Universitari Vall d'Hebron, 08035 Barcelona, Spain Department of Cell Biology,⁵ Georgetown University Medical Center, Washington, DC 20007

Progression of the first meiotic division in male germ cells is regulated by a variety of factors, including androgens and possibly estrogens. When this regulation fails, meiosis is arrested and primary spermatocytes degenerate by apoptosis. Earlier studies showed that overexpression of rat androgen-binding protein (ABP) in the testis of transgenic mice results in a partial meiotic arrest and apoptosis of pachytene spermatocytes. In view of the recent localization of estrogen receptor ß (ERß) in primary spermatocytes and data suggesting the ability of ERß to repress cellular proliferation, we tested the hypothesis that variations in the testicular steroid microenvironment caused by excess ABP produce changes in ERß expression in this cellular type that could be associated to the meiotic arrest and, eventually, to the induction of germ cell apoptosis observed in the ABP transgenic mice. Increased levels of ERß mRNA and protein were demonstrated in the testis of rat ABP transgenic mice compared with nontransgenic littermates by reverse transcriptase-polymerase chain reaction (RT-PCR) experiments, Northern blotting, and Western Blotting. The major differences were found when isolated germ cells of transgenic and nontransgenic littermates were analyzed by RT-PCR. In keeping with this finding, ERß was strongly immunolabeled in pachytene spermatocytes of rat ABP transgenic mice and localized in tubular stages in which TUNEL labeling was maximal. Confocal microscopy analysis of a fluorescent TUNEL assay and ERß immunohistochemistry revealed that degenerating pachytene spermatocytes overexpressed ERß. The present results are consistent with the interpretation that ERß is associated with the events that regulate negatively the progression of meiosis or that lead to spermatocyte apoptosis.

¹ Support: This work was funded by the Ministerio de Educación y Ciencia (PM99/0134 y acción integrada con Francia HI999-0067), Ministerio de Sanidad y Consumo (98/0354), Serono-Fundación Salud 2000 y Comisionat de Recerca-CIRIT (1999/SGR00231).

² Correspondence: Francina Munell, Grup de Recerca en Endocrinologia Molecular, Hospital Materno-Infantil, planta 14, Hospital Universitari Vall d'Hebrón, Pg. Vall d'Hebron, 119-129, 08035 Barcelona, Spain. FAX: 34 934894064; fmunell{at}vhebron.net

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