Multiple structurally distinct ERα mRNA variants in zebrafish are differentially expressed by tissue type, stage of development and estrogen exposure.
Abstract
It is well established that estrogen-like environmental chemicals interact with the ligand-binding site of estrogen receptors (ERs) to disrupt transcriptional control of estrogen responsive targets. Here we investigate the possibility that estrogens also impact splicing decisions on estrogen responsive genes, such as that encoding ERα itself. Targeted PCR cloning was applied to identify six ERα mRNA variants in zebrafish. Sequencing revealed alternate use of transcription and translation start sites, multiple exon deletions, intron retention and alternate polyadenylation. As determined by quantitative (q)PCR, N-terminal mRNA variants predicting long (ERαA(L)) and short (ERα(S)) isoforms were differentially expressed by tissue-type, sex, stage of development and estrogen exposure. Whereas ERα(L) mRNA was diffusely distributed in liver, brain, heart, eye, and gonads, ERα(S) mRNA was preferentially expressed in liver (female>male) and ovary. Neither ERα(L) nor ERα(S) transcripts varied significantly during development, but 17β-estradiol selectively increased accumulation of ERα(S) mRNA (∼170-fold by 120 hpf), an effect mimicked by bisphenol-A and diethylstilbestrol. Significantly, a C-truncated variant (ERα(S)-Cx) lacking most of the ligand binding and AF-2 domains was transcribed exclusively from the short isoform promoter and was similar to ERα(S) in its tissue-, stage- and estrogen inducible expression. These results support the idea that promoter choice and alternative splicing of the esr1 gene of zebrafish are part of the autoregulatory mechanism by which estrogen modulates subsequent ERα expression, and further suggest that environmental estrogens could exert some of their toxic effects by altering the relative abundance of structurally and functionally distinct ERα isoforms.
Keywords
bisphenol a
estrogen receptor
zebrafish
race
endocrine disrupting chemicals
qpcr
er
dimethyl sulfoxide
dmso
17β-estradiol
endocrine disruption
bpa
af
alternative splicing
dbd
des
dna binding domain
e2
edc
estrogen receptor alpha
lbd
tcdd
tss
xe
activation function
diethylstilbestrol
hours post fertilization
hpf
ligand binding domain
mrna variants
quantitative polymerase chain reaction
rapid amplification of cdna ends
tetrachlorodibenzo-p-dioxin
transcription start site
xenoestrogens
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74040
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Article ID:
74040
Unique Identifier:
10.1016/j.ygcen.2013.09.014
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Scimatic Chain (ID: 481)
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