Isolation and characterization of the cDNA encoding the tilapia (Oreochromis niloticus) cytochrome P450 aromatase (P450arom): changes in P450arom mRNA, protein and enzyme activity in ovarian follicles during oogenesis.
A cDNA clone encoding the complete tilapia (a teleost fish, Oreochromis niloticus) cytochrome P450 aromatase (P450arom) was isolated from an ovarian follicle cDNA library. The deduced amino acid sequence (522 amino acid residues) had 72.2% and 59.5% homology with rainbow trout and catfish P450arom respectively, and about 50% homology with mammalian and avian P450arom. Expression of this cDNA in COS-7 cells produced a protein that converted exogenous testosterone to estrogens. Northern blots using a tilapia P450arom cDNA fragment and Western blots using an antiserum against a tilapia P450arom polypeptide fragment revealed a single P450arom mRNA (2.6 kb) and a single protein (59 kDa) in tilapia ovarian tissue respectively. These analyses also revealed that the levels of both P450arom mRNA and protein were low in early vitellogenic follicles, increased in midvitellogenic follicles, and declined to non-detectable levels in post-vitellogenic follicles. Changes in the ability of follicles to convert exogenous testosterone to estrogens (aromatase activity) were similar to those of P450arom mRNA and protein. These observations indicated that the capacity of tilapia ovarian follicles to synthesize estradiol-17 beta is closely related to the contents of P450arom mRNA and protein within them.
Chang
X T
XT
Laboratory of Reproductive Biology, National Institute for Basic Biology, Okazaki, Japan.
Kobayashi
T
T
Kajiura
H
H
Nakamura
M
M
Nagahama
Y
Y
D000595:Amino Acid Sequence; D000818:Animals; D001141:Aromatase / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D019556:COS Cells; D018076:DNA, Complementary; D005260:Female; D008969:Molecular Sequence Data; D009866:Oogenesis; D006080:Ovarian Follicle / Q000201:enzymology*; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D017210:Tilapia / Q000235:genetics*
D018076:DNA, Complementary; D012333:RNA, Messenger; D001141:Aromatase