Cloning and gene expression of P450 17alpha-hydroxylase,17,20-lyase cDNA in the gonads and brain of the fathead minnow Pimephales promelas.
P450 17alpha-hydroxylase,17,20-lyase (P450c17) is a key steroidogenic enzyme in the production of androgens and, therefore, is also indispensable for the production of oestrogens (that are produced from the aromatisation of androgens). In this study, P450c17 cDNA was cloned from the ovary of the fathead minnow (FHM) and its gene expression was examined in the gonads and brains of male and female FHM at different stages of gonadal development with a view to developing an understanding of its involvement in the reproductive physiology in this species. The FHM-P450c17 cDNA sequence cloned was 1812 bp in length, with an open reading frame of 1554 nucleotides encoding a protein of 518 amino acids. Amino acid identity of FHM-P450c17 with P450c17s in other animals was up to 81.8% in other teleosts (channel catfish), 62% in elasmobranches (spiny dogfish), 64% in birds (chicken), and up to 48.8% in mammals (human). FHM-P450c17 gene expression occurred in the ovary, testis, and also in the brain (both male and female) at all stages of sexual development studied. Expression in the brain was at least 30-fold lower than in the gonads, but consistent in all fish life stages studied. In the testis, FHM-P450c17 gene expression was negatively correlated with gonadal development, but there was no obvious association between P450c17 gene expression and sexual development in the ovary, or brain (in both males and females). The results from this study demonstrate the expression of P450c17 in the brain for the first time in fish. Enzymatic studies are now needed to investigate the possible role of P450c17 in neurosteroid production in teleosts.
Halm
S
S
Instituto de Acuicultura de Torre de la Sal, Ribera de Cabanes, 12595 Torre de la Sal (Castellón), Spain. silkehalm@iats.csic.es
Kwon
J Y
JY
Rand-Weaver
M
M
Sumpter
J P
JP
Pounds
N
N
Hutchinson
T H
TH
Tyler
C R
CR
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000201:enzymology*; D003001:Cloning, Molecular; D003530:Cyprinidae / Q000235:genetics* / Q000378:metabolism; D018076:DNA, Complementary / Q000032:analysis*; D005260:Female; D005786:Gene Expression Regulation; D006066:Gonads / Q000201:enzymology* / Q000254:growth & development; D016014:Linear Models; D008297:Male; D008969:Molecular Sequence Data; D012098:Reproduction / Q000235:genetics / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D012737:Sex Factors; D013254:Steroid 17-alpha-Hydroxylase / Q000096:biosynthesis / Q000235:genetics*
D018076:DNA, Complementary; D013254:Steroid 17-alpha-Hydroxylase