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First published online September 19, 2006
Journal of Experimental Biology 209, 3806-3811 (2006)
Published by The Company of Biologists 2006
doi: 10.1242/jeb.02424

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The expression level of frog relaxin mRNA (fRLX), in the testis of Rana esculenta, is influenced by testosterone

Gianluca De Rienzo¹, Francesco Aniello², Margherita Branno³, Gaia Izzo¹ and Sergio Minucci^1,*

¹ Dipartimento di Medicina Sperimentale, Sez. Fisiologia Umana e Funzioni Biologiche Integrate `F. Bottazzi', Seconda Università di Napoli, via Costantinopoli 16, 80138 Napoli, Italy
<sup>2</sup> Dipartimento di Biologia Strutturale e Funzionale, Università di Napoli `Federico II', via Cinthia, 80126 Napoli, Italy
³ Laboratorio di Biochimica e Biologia Molecolare, Stazione Zoologica `A. Dohrn', Villa Comunale, 80121 Napoli, Italy

^* Author for correspondence (e-mail: sergio.minucci{at}unina2.it)

Accepted 3 July 2006

Frog relaxin (fRLX) belongs to the relaxin/insulin gene family present in the testis of Rana esculenta and is specifically expressed by Leydig cells. Since the expression of fRLX transcript changes during the reproductive cycle and is more abundant when circulating levels of androgens are relatively high, we investigated the effect(s) of testosterone and its antagonist (cyproterone acetate, CPA) on its expression pattern, in the testis of the frog Rana esculenta. Results from in vivo and in vitro experiments demonstrate that testosterone strongly induces a significant increase of fRLX mRNA expression in frog testes and, this effect is counteracted by CPA, supporting the existence of intratesticular (autocrine/paracrine) mechanisms of action. Interestingly, in both the control and testosterone-treated testes, fRLX mRNA expression was markedly decreased 24 h post-treatment, as compared to that measured at 2 h and 8 h post-treatment, suggesting that factor(s), other than testosterone, may act(s) in controlling its expression. In addition, RT-PCR analysis and in situ hybridization performed on frog testis injected with CPA for 15 days, on alternate days, showed a strong decrease of fRLX expression, suggesting that CPA counteracts the effect of testosterone on fRLX expression. Taken together our results strongly indicate that changes in the production, by the Leydig cells, of both testosterone and fRLX may represent a marker for the study of Leydig cell activity in the testis of the frog Rana esculenta.

Key words: relaxin, testosterone, cyproterone acetate, gene expression, Rana esculenta

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