Cloning and expression of P450c17-I (17α-hydroxylase/17,20-lyase) in brain and ovary during gonad development in Cynoglossus semilaevis.
Cytochrome P450c17 (CYP17, 17a-hydroxylase/17,20-lyase) is a critical enzyme in the production of androgens and estrogens in vertebrates. A 2,469 bp full length cDNA of P450c17-I (CYP17A1) has been isolated from the ovary of half-smooth tongue sole, Cynoglossus semilaevis which encodes 509 amino acids. Additionally, a relatively shorter cDNA (1,742 bp), a likely result of polyadenylation, was also found. The putative P450c17-I enzyme shares high sequence identity with that of the fathead minnow (73%), zebrafish (71%), the Japanese eel (70%), catfish (70%), tilapia (79%), three-spined stickleback (81%), medaka (79%), dogfish (60%), chicken (65%), rat (47%), and human (49%). Semi-quantitative RT-PCR analysis of spatial expression showed the enzyme was predominantly expressed in the ovaries and the brain. P450c17-I was also detected in the stomach, intestine, gill, spleen, kidney, and head kidney, albeit weakly. Further examination of temporal expression pattern of P450c17-I in ovary and brain revealed developmental stage-dependency. In addition to this our data on T and E2 levels further endorse the critical role of P450c17-I during shift in steroidogenesis. Based on the present study we indicate an important role for P450c17-I during ovarian development. However, further studies are needed at transcriptional regulation level for deeper insights into the physiological functions of P450c17-I.
Chen
Cai F
CF
Fisheries College, Ocean University of China, 5 Yushan Road, 266003, Qingdao, China.
Wen
Hai S
HS
Wang
Zhao P
ZP
He
Feng
F
Zhang
Jia R
JR
Chen
Xiao Y
XY
Jin
Guo X
GX
Shi
Bao
B
Shi
Dan
D
Yang
Yan P
YP
Li
Ji F
JF
Qi
Bao X
BX
Li
Na
N
D000595:Amino Acid Sequence; D000704:Analysis of Variance; D000818:Animals; D001921:Brain / Q000201:enzymology*; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D004958:Estradiol / Q000097:blood; D005260:Female; D005412:Flatfishes / Q000235:genetics*; D020869:Gene Expression Profiling; D006652:Histological Techniques; D008969:Molecular Sequence Data; D010053:Ovary / Q000201:enzymology* / Q000254:growth & development; D011863:Radioimmunoassay; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D017385:Sequence Homology; D013045:Species Specificity; D013254:Steroid 17-alpha-Hydroxylase / Q000235:genetics* / Q000378:metabolism*; D013739:Testosterone / Q000097:blood
D017931:DNA Primers; D018076:DNA, Complementary; D013739:Testosterone; D004958:Estradiol; D013254:Steroid 17-alpha-Hydroxylase