Induction of phase II enzymes and hsp70 genes by copper sulfate through the electrophile-responsive element (EpRE): insights obtained from a transgenic zebrafish model carrying an orthologous EpRE sequence of mammalian origin.
We have evaluated the homology of the electrophile-responsive element (EpRE) core sequence, a binding site for the Nrf2 transcription factor, in the proximal promoters of the mouse and zebrafish glutathione-S-transferase (gst), glutamate cysteine ligase catalytic subunit (gclc) and heat shock protein 70 (hsp70) genes. The EpRE sites identified for both species in the three analyzed genes showed a high similarity with the putative EpRE core sequence. We also produced a transgenic zebrafish model carrying a transgene comprised of the luciferase (luc) reporter gene under transcriptional control of a mouse EpRE sequence. This transgenic model was exposed to copper sulfate, and the reporter gene was significantly activated. The endogenous gst, gclc and hsp70 zebrafish genes were analyzed in the EpRE-Luc transgenic zebrafish and showed an expression pattern similar to that of the reporter transgene used. Our results demonstrate that EpRE is conserved between mouse and zebrafish for detoxification-related genes and that the development of genetically modified models using this responsive element to drive the expression of reporter genes can be an important tool in understanding the action mechanism of aquatic pollutants.
Almeida
Daniela Volcan
DV
Programa de Pós-graduação em Ciências Fisiológicas, Fisiologia Animal Comparada, Instituto de Ciências Biológicas, Universidade Federal do Rio Grande - FURG, Av. Itália km 8, CEP 96201-900, Rio Grande, RS, Brazil.
da Silva Nornberg
Bruna Félix
BF
Programa de Pós-graduação em Ciências Fisiológicas, Fisiologia Animal Comparada, Instituto de Ciências Biológicas, Universidade Federal do Rio Grande - FURG, Av. Itália km 8, CEP 96201-900, Rio Grande, RS, Brazil.
Geracitano
Laura A
LA
Programa de Pós-graduação em Ciências Fisiológicas, Fisiologia Animal Comparada, Instituto de Ciências Biológicas, Universidade Federal do Rio Grande - FURG, Av. Itália km 8, CEP 96201-900, Rio Grande, RS, Brazil.
Barros
Daniela Martí
DM
Programa de Pós-graduação em Ciências Fisiológicas, Fisiologia Animal Comparada, Instituto de Ciências Biológicas, Universidade Federal do Rio Grande - FURG, Av. Itália km 8, CEP 96201-900, Rio Grande, RS, Brazil.
Monserrat
José Maria
JM
Programa de Pós-graduação em Ciências Fisiológicas, Fisiologia Animal Comparada, Instituto de Ciências Biológicas, Universidade Federal do Rio Grande - FURG, Av. Itália km 8, CEP 96201-900, Rio Grande, RS, Brazil.
Marins
Luis Fernando
LF
Programa de Pós-graduação em Ciências Fisiológicas, Fisiologia Animal Comparada, Instituto de Ciências Biológicas, Universidade Federal do Rio Grande - FURG, Av. Itália km 8, CEP 96201-900, Rio Grande, RS, Brazil. dqmluf@furg.br.
D000818:Animals; D030801:Animals, Genetically Modified; D020134:Catalytic Domain / Q000235:genetics; D017124:Conserved Sequence / Q000235:genetics; D019327:Copper Sulfate / Q000633:toxicity*; D017931:DNA Primers / Q000235:genetics; D005786:Gene Expression Regulation / Q000187:drug effects*; D005721:Glutamate-Cysteine Ligase / Q000378:metabolism*; D005982:Glutathione Transferase / Q000378:metabolism*; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism*; D008658:Inactivation, Metabolic / Q000235:genetics / Q000502:physiology; D051379:Mice; D051267:NF-E2-Related Factor 2 / Q000235:genetics / Q000378:metabolism*; D020218:Response Elements / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015027:Zebrafish
D017931:DNA Primers; D018840:HSP70 Heat-Shock Proteins; D051267:NF-E2-Related Factor 2; D005982:Glutathione Transferase; D005721:Glutamate-Cysteine Ligase; D019327:Copper Sulfate