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Molecular Biology of Ovarian Aromatase in Sex Reversal: Complementary DNA and 5'-Flanking Region Isolation and Differential Expression of Ovarian Aromatase in the Gilthead Seabream (Sparus aurata)¹

Center of Marine Biotechnology, University of Maryland Biotechnology Institute, Baltimore, Maryland 21202

ABSTRACT

To elucidate the involvement of aromatase in sex reversal, the gilthead seabream ovarian P450 aromatase (cyp19a1a) cDNA and its 5'-flanking region were isolated and characterized. Northern blot analysis revealed that only one cyp19a1a transcript (2.0 kb) is expressed in the ovary. Four cAMP-responsive elements were identified at the 5'-flanking region of seabream cyp19a1a indicating a high potential to respond to gonadotropin signaling. Studying the seasonal profile, two expression peaks of cyp19a1a transcripts in the ovarian tissues were found in July (about 15000 copies/ng total RNA) for ambisexual fish and in December (about 12000 copies/ng total RNA) for spawning females. Starting from September, transcript levels of cyp19a1a in the ovarian portions of the male-developing gonads gradually decreased. Furthermore, the ovarian portions of the female gonads expressed cyp19a1a at a significantly higher level than the ovarian portions of the male gonads after November. Taken together with levels of plasma estradiol in reversing females being significantly higher than those in developing males, the above results reinforce the importance of cyp19a1a in sex reversal. In vitro exposure of ovarian fragments to gonadotropins (hCG) at 1, 10, and 100 IU/ml significantly (P < 0.05) upregulated cyp19a1a expression. Additionally, expression of cyp19a1a displayed a stronger and significant correlation with the transcript expression of ovarian Lh receptor rather than Fsh receptor during the ambisexual stage. Our results indicate that the differential expression of cyp19a1a gene is associated with sex reversal and that gonadotropin signals (particularly Lh) may serve as major players in regulating the expression of cyp19a1a during the process of sex reversal.

aromatase, estradiol, follicle-stimulating hormone, luteinizing hormone, ovary, sex reversal

¹ Supported by a Maryland Sea Grant Fellowship to T.-T.W. and award NA46RG0091 to Y.Z. Contribution 05–127 from the Center of Marine Biotechnology, University of Maryland Biotechnology Institute.

² Correspondence: Yonathan Zohar, 701 E. Pratt St., Baltimore, MD 21202. FAX: 410 234 8896; zohar{at}umbi.umd.edu

³ Current address: Department of Animal Sciences, Purdue University, West Lafayette, IN 47907.

⁴ Current address: Department of Reproductive Biology, National Institute of Basic Biology, Nishigonaka 38, Myodaiji, Okazaki 444-8585 Aichi, Japan.

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