BOR - Papers in Press, published online ahead of print
November 27, 2002.
Biol Reprod 2002, 10.1095/biolreprod.102.010843
BIOLOGY OF REPRODUCTION 68, 15451553 (2003)
DOI: 10.1095/biolreprod.102.010843
© 2003 by the Society for the Study of Reproduction, Inc.
Synergistic Expression of Ad4BP/SF-1 and Cytochrome P-450 Aromatase (Ovarian Type) in the Ovary of Nile Tilapia, Oreochromis niloticus, During Vitellogenesis Suggests Transcriptional Interaction1
Yasutoshi Yoshiura3,4,
Balasubramanian Senthilkumaran3,7,
Masatada Watanabe3,5,
Yuichi Oba3,6,
Tohru Kobayashi3,7, and
Yoshitaka Nagahama2,3,7
Laboratory of Reproductive Biology,3 National Institute for Basic Biology, Okazaki 444-8585, Japan
Immunology Section,4 Inland Station, National Research Institute of Aquaculture, Tamaki, Mie 519-0423, Japan
Department of Biochemistry,5 Faculty of Pharmaceutical Sciences, Hoshi University, Shinagawa-Ku, Tokyo 142-0063, Japan
Graduate School of Bioagricultural Sciences,6 Nagoya University, Nagoya 464-8601, Japan
CREST,7 Japan Science and Technology Corporation, Kawaguchi, Saitama 332-0012, Japan
Involvement of Ad4BP/SF-1 in the ovarian cytochrome P-450 aromatase (oP450arom) gene expression was investigated using ovarian follicles of the Nile tilapia, possessing an average 14-day spawning cycle. The promoter region (5' flanking region) of oP450arom gene cloned from tilapia contains two Ad4 binding sites. Subsequently, a cDNA encoding Ad4BP/SF-1 was cloned from the ovarian follicles. It is expressed in gonadal tissues, brain, and kidney. Oligonucleotide probes containing putative orphan nuclear receptor binding motifs (derived from promoter region of the aromatase gene) formed complexes with in vitro-translated Ad4BP/SF-1 and nuclear extracts of tilapia ovarian (midvitellogenic) follicles, indicating that Ad4BP/SF-1 is one of the transcriptional regulators for aromatase gene expression. Northern blot analysis revealed that the expression of both oP450arom and Ad4BP/SF-1 increased in parallel with ovarian growth from Day 0 to Day 5 after spawning and declined sharply from Day 8 to Day 11. On the day of spawning (Day 14), the expression of both correlates became undetectable. In vitro incubation of post vitellogenic full-grown immature follicles (corresponding to Day 11 after spawning) with hCG purged both oP450arom and Ad4BP/SF-1 messenger RNA transcripts at 18 h. Conversely, in vitro incubation of late vitellogenic follicles (corresponding to Day 8 after spawning) with hCG retained Ad4BP/SF-1 messenger RNA transcripts more or less steadily and up-regulated oP450arom. Ad4BP/SF-1 probably acts as a transcriptional modulator to implement the paradoxical actions of gonadotropins on oP450arom gene.
1 Supported in part by Grants-in-Aid for Research for the Future (JSPS-RFTF 96L100401) and Priority Areas (07283101) from the Japan Society for the Promotion of Science; for Scientific Research from the Ministry of Education, Science, Sports, and Culture, Japan; Bio-Design Program from the Ministry of Agriculture, Forestry, and Fisheries, Japan; and CREST of JST (Japan Science and Technology).
2 Correspondence: Yoshitaka Nagahama, National Institute for Basic Biology, Okazaki 444-8585, Japan. FAX: 81 564 55 7556; nagahama{at}nibb.ac.jp
Copyright © 2003 by the Society for the Study of Reproduction.