Characterization of a cDNA encoding P-450 aromatase (CYP19) from Japanese eel ovary and its expression in ovarian follicles during induced ovarian development.
A cDNA encoding P450 aromatase (CYP19) was isolated from a Japanese eel (Anguilla japonica) ovarian cDNA library. This cDNA contains a complete open reading frame encoding 511 amino acid residues. The deduced amino acid sequence is 59% and 65% identical to the catfish and rainbow trout forms, respectively, and 52-54% to mammalian and chicken forms. Non-steroidogenic COS-7 cells transfected with the eel CYP19 cDNA converted exogenous androstenedione to estrone, thus verifying its identity. Northern blot analysis indicated that there was a single 2.1 kb transcript in the ovary. A 2.1 kb transcript was also found in the brain but not in the spleen, head kidney, kidney, or liver. Throughout ovarian development induced by weekly injections of salmon pituitary homogenate (SPH, 20 microg/g body weight), the 2.1 kb transcript was barely or not detectable in the ovaries. However, signals greatly increased in intensity in oocytes in the migratory nucleus stage and then decreased slightly in the post-ovulatory ovary. These changes in transcript levels are consistent with the changes in aromatase activity of ovarian follicles, suggesting that aromatase activity in ovarian follicles is mainly regulated at the transcriptional level. In addition, fadrozole was found to significantly inhibit aromatase activity in a heterologous expression system using COS-7 cells, which indicates that fadrozole treatment could be useful to control E(2) production during artificial maturation of eels.
Ijiri
Shigeho
S
Division of Marine Biosciences, Graduate School of Fisheries Science, Hokkaido University, 3-1-1 Minato-cho, Hakodate, Hokkaido 041-8611, Japan. ijiri@kun.ne.jp
Kazeto
Yukinori
Y
Lokman
P Mark
PM
Adachi
Shinji
S
Yamauchi
Kohei
K
D000595:Amino Acid Sequence; D000811:Anguilla / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D001141:Aromatase / Q000096:biosynthesis* / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000302:isolation & purification*; D005260:Female; D008969:Molecular Sequence Data; D006080:Ovarian Follicle / Q000737:chemistry / Q000201:enzymology* / Q000254:growth & development; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis; D017385:Sequence Homology; D014818:Vitellogenesis / Q000502:physiology*
D018076:DNA, Complementary; D012333:RNA, Messenger; D001141:Aromatase