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Distinct cytochrome P450 aromatase isoforms in zebrafish (Danio rerio) brain and ovary are differentially programmed and estrogen regulated during early development.
- Kishida M, Callard GV
Endocrinology. 2001 Feb;142(2):740-50.
As a first step toward understanding estrogen's
role
in neurodevelopment, a PCR
cloning
strategy
was used to isolate
complementary
DNAs encoding two distinct cytochrome P450
aromatase
isoforms in adult zebrafish
(Danio rerio) brain and ovary (termed P450aromB and P450aromA, respectively). Sequence and phylogenetic
analysis
showed that the zebrafish
P450arom forms are orthologs of previously identified cyp19b and cyp19a genes
in goldfish.
On Northern blots,
a single 4.4-kb transcript of the P450aromB subtype was identified in brain, and a 2.1-kb transcript of the P450aromA subtype in ovary, but RT-PCR showed a degree of overlapping
expression. Both messenger RNA
(mRNA)
forms were detected in unfertilized eggs and 1.5 hpf (cleavage
stage) embryos but declined by 12 hpf, indicating maternal transfer.
A secondary rise in mRNAs between 12-24 hpf indicated the onset of embryonic cyp19b and -a transcription. Both mRNA
species accumulated progressively to 120 hpf (early larval stage), but the relative magnitude
and pattern of change was isoform
specific. Estradiol
(E(2,) 1 microM) advanced and amplified the developmentally programmed accumulation of P450aromB mRNA,
and ICI164.384 decreased expressed levels, implying blockade of an endogenous estrogen
mediated regulatory component.
Conversely, E(2) had no effect or decreased P450aromA mRNA.
The early embryonic expression of P450aromB and P450aromA isoforms, and differences in developmental programming and estrogen
regulation,
imply independent regulatory mechanisms and unique functions during major morphogenetic and differentiative events.
This abstract at PubMed.