Molecular Identification OfKiss/GPR54 And Function Analysis With M RNA Expression Profiles Exposure To 17α-ethinylestradiol In Rare Minnow Gobiocypris Rarus

17α-Ethinylestradiol(EE2), one of environmental endocrine disruptors, is a most potent estrogens and the basis of artificial birth control pills. EE2 absolutely impacts the health of human and animals when it accumulated in the water. These effects include eliciting reproductive failure, disrupting the endocrine, retarding gonadal growth and altering sexual differentiation. In the present study, we aim to study the disruption on EE2 for Kiss/GPR54 genes expression and screen sensitive biomarkers for estrogenic chemicals in the environmental monitoring. The gene expression of Kiss/GPR54 detected in brain and gonads in rare minnow will benefit for exploring the mechanism underlying the impact of EE2 on reproduction and gonadal development through neuroendocrine system.In the present study, the full length c DNA sequences of kiss1, kiss2, GPR54 a and GPR54 b were isolated in rare minnow using RT-PCR and RACE methods. The tissue distribution of these four genes were detected inseven tissues including brain, eye, gill, liver, intestine, muscle and testis or ovary by q RT-PCR. Then, the adult rare minnow at dpf 105 were exposed to EE2 at 1, 5, 25 and 125 ng/L for 3 and 6 days. The gene expressions of kiss1, kiss2, GPR54 a,GPR54b, Gn RH2 and Gn RH3 were detected in brain and kiss2, GPR54 a and GPR54 b were detected in gonads of rare minnow by q RT-PCR. The main results were obtained as follow:1. The full length cDNAs of kiss1, kiss2, GPR54 a and GPR54 b were 794, 628, 1785 and 1624 bp, respectively. The open reading frames were 351, 381, 1098 and 1116 bp, encoding 116, 126, 365 and 371 amino acids. The putative amino acid sequences for these four genes shared the identical function-elements and structural domains of their respective counterparts inthe close teleost species.2. The highexpression of Kiss/GPR54 were detected in brain and gonad in rare minnow.. 3. The expression of kiss1 and kiss2 were significantly increased by EE2 in female brain. However the GPR54 b was significantly inhibited in gonads of both sexes.4. The four genes cloned in the present study can be used as sensitive biomarkers to detect the estrogenic chemicals for the environmental monitoring. The genes of kiss2 and GPR54 b rather than kiss1 and GPR54 a played the most important role in regulatingreproduction and development inresponse to EE2 disruption.