[Gene cloning, sequence analysis and tissue expression of estrogen-related receptor alpha (Erralpha) in Japanese medaka and its transcriptional responses after differential EDCs exposure].
Endocrine disrupting chemicals (EDCs) can bind or block nuclear receptors in the body and subsequently affect growth, development and reproduction of fish. Estrogen-related receptors (ERRs), belonging to the nuclear receptor superfamily, have been implicated in diverse physiological processes in estrogen signal pathway in mammals, while little is known about them in fishes. Complete mRNA sequence of ERRalpha from medaka (Oryzias latipes) was cloned, and the sequence is similar to those of other vertebrates, especially that the DNA-binding domain (DBD) of ERRalpha is highly conserved among the vertebrates (97.4%-100% sequence identities) and the ligand-binding domain (LBD) of medaka ERRalpha is 66.4%-67.0% sequence identities with those of mammals. The DBD of medaka ERRalpha is of the same length and has high sequence identity with those of estrogen receptor (ERalpha and ERbeta) and androgen receptor (ARalpha and ARbeta) of medaka, but much difference was found between the LBD of medaka ERRalpha with those of ERalpha, ERbeta, ARalpha and ARbeta. ERRalpha gene is located in chromosome 14 and is consisted of 5 exons. The expressions of ERRalpha in different tissues and the transcriptional responses of ERRalpha in testis of medaka exposed differential EDCs were studied by quantitative real-time RT-PCR. ERRalpha is expressed at apparently high levels in gonad, brain, eye, spleen and intestine, though it was broadly expressed in tissues. Significant transcriptional difference was found between testis and ovary, implying ERRalpha would be involved in sex differentiation and gonad development in fish. After 3 weeks exposure of medaka to 200 ng/L ethynylestradiol (EE2), 200 ng/L estrone (E1), 200 ng/L diethylstilbestrol (DES), 100 microg/L atrazine (AT) and 200 ng/L 17beta-estradiol (E2), transcripts of ERRalpha were significantly decreased to 0.54, 0.56, 0.61, 0.63 and 0.65 of control (p < 0.05) in the testes, respectively. And those in the 1 microg/L tributyltin (TBT) and 1 microg/L triphenyltin (TPT) exposure groups were up-regulated to 1.34 and 1.35 folds of control (p > 0.05), respectively. These results suggested that ERRalpha would take actions in the disruption of sex differentiation and gonad development in fish by EDCs. In addition, no multiple steroid hormone-response element half-sites was found in medaka, which were reported in the upstream of ERRalpha gene in mammals, indicating there would be different regulation patters of ERRalpha between teleost and mammal.
Zhang
Zhao-Bin
ZB
College of Urban and Environmental Sciences, Peking University, Beijing 100871, China. zhangzb@urban.pku.edu.cn
Hu
Jian-Ying
JY
Sai
Si-Xiang
SX
Zhao
Yan-Bin
YB
Huang
Chong
C
Tian
Xiao-Jun
XJ
D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D052244:Endocrine Disruptors / Q000633:toxicity*; D004781:Environmental Exposure; D004785:Environmental Pollutants / Q000633:toxicity; D008969:Molecular Sequence Data; D009990:Oryzias / Q000235:genetics* / Q000378:metabolism; D011960:Receptors, Estrogen / Q000235:genetics / Q000378:metabolism*; D017423:Sequence Analysis, RNA; D014158:Transcription, Genetic; D000097983:ERRalpha Estrogen-Related Receptor
D052244:Endocrine Disruptors; D004785:Environmental Pollutants; D011960:Receptors, Estrogen; D000097983:ERRalpha Estrogen-Related Receptor