Several cohort studies have reported that dioxin and dioxin-like polychlorinated biphenyls might impair the nervous system and lead to neurological or neurodegenerative diseases in the elder people, but there is limit...Several cohort studies have reported that dioxin and dioxin-like polychlorinated biphenyls might impair the nervous system and lead to neurological or neurodegenerative diseases in the elder people, but there is limited research on the involved mechanism. By using microarray analysis, we figured out the differentially expressed genes between brain samples from SD rats after low-dose（0.1 μg/（kg？bw）） dioxin exposure for six months and controls. To investigate the function changes in the course of dioxin exposure, Gene Ontology（GO） annotation and Kyoto Encyclopedia of Genes and Genomes（KEGG） pathway analysis were performed on the differentially expressed genes. And the changes of several picked genes have been verified by real-time PCR. A total of 145 up-regulated and 64 down-regulated genes were identified. The metabolic processes, interleukin-1 secretion and production were significantly associated with the differentially expressed genes. And the genes regulated by dioxin also clustered to cholinergic synapse and long-term potentiation. Candidate biomarker genes such as egr1, gad2, gabrb3, abca1, ccr5 and pycard may be toxicological targets for dioxin. Furthermore, synaptic plasticity and neuro-immune system may be two principal affected areas by dioxin.展开更多
Aryl hydrocarbon receptor（Ah R）, a ligand-dependent nuclear receptor, is involved in a diverse spectrum of biological and toxicological effects. Due to the lack of three dimensional（3D）crystal or nuclear magnetic ...Aryl hydrocarbon receptor（Ah R）, a ligand-dependent nuclear receptor, is involved in a diverse spectrum of biological and toxicological effects. Due to the lack of three dimensional（3D）crystal or nuclear magnetic resonance structure, the mechanisms of these complex effects of AhR remain to be unclear. Also, commercial monoclonal antibodies（mA bs） against human AhR protein（h Ah R）, as alternative immunological tools, are very limited. Thus, in order to provide more tools for further studies on h Ah R, we prepared two m Abs（1D6 and 4A6） against h Ah R. The two newly generated m Abs specifically bound to amino acids 484–508（located in transcription activation domain） and amino acids 201–215（located in Per-ARNT-Sim domain）of h Ah R, respectively. These epitopes were new as compared with those of commercial m Abs.The m Abs were also characterized by enzyme-linked immunosorbent assay, western blot,immunoprecipitation and indirect immunofluorescence assay in different cell lines. The results showed that the two m Abs could recognize the linearized AhR s in six different human cell lines and a rat hepatoma cell line, as well as the h Ah R with native conformations. We concluded that the newly generated m Abs could be employed in AhR-based bioassays for analysis of environmental contaminants, and held great potential for further revealing the spatial structure of AhR and its biological functions in future studies.展开更多
基金supported by grants from the Natural Science Foundation of China(Nos.21407171,21377160,21525730)the Strategic Priority Research Program of the Chinese Academy of Sciences(No.XDB14030400)
文摘Several cohort studies have reported that dioxin and dioxin-like polychlorinated biphenyls might impair the nervous system and lead to neurological or neurodegenerative diseases in the elder people, but there is limited research on the involved mechanism. By using microarray analysis, we figured out the differentially expressed genes between brain samples from SD rats after low-dose（0.1 μg/（kg？bw）） dioxin exposure for six months and controls. To investigate the function changes in the course of dioxin exposure, Gene Ontology（GO） annotation and Kyoto Encyclopedia of Genes and Genomes（KEGG） pathway analysis were performed on the differentially expressed genes. And the changes of several picked genes have been verified by real-time PCR. A total of 145 up-regulated and 64 down-regulated genes were identified. The metabolic processes, interleukin-1 secretion and production were significantly associated with the differentially expressed genes. And the genes regulated by dioxin also clustered to cholinergic synapse and long-term potentiation. Candidate biomarker genes such as egr1, gad2, gabrb3, abca1, ccr5 and pycard may be toxicological targets for dioxin. Furthermore, synaptic plasticity and neuro-immune system may be two principal affected areas by dioxin.
基金supported by the National Natural Science Foundation of China (Nos. 21277168, 21525730)the Strategic Priority Research Program of the Chinese Academy of Sciences (Nos. XDB14030401, XDB14030402)
文摘Aryl hydrocarbon receptor（Ah R）, a ligand-dependent nuclear receptor, is involved in a diverse spectrum of biological and toxicological effects. Due to the lack of three dimensional（3D）crystal or nuclear magnetic resonance structure, the mechanisms of these complex effects of AhR remain to be unclear. Also, commercial monoclonal antibodies（mA bs） against human AhR protein（h Ah R）, as alternative immunological tools, are very limited. Thus, in order to provide more tools for further studies on h Ah R, we prepared two m Abs（1D6 and 4A6） against h Ah R. The two newly generated m Abs specifically bound to amino acids 484–508（located in transcription activation domain） and amino acids 201–215（located in Per-ARNT-Sim domain）of h Ah R, respectively. These epitopes were new as compared with those of commercial m Abs.The m Abs were also characterized by enzyme-linked immunosorbent assay, western blot,immunoprecipitation and indirect immunofluorescence assay in different cell lines. The results showed that the two m Abs could recognize the linearized AhR s in six different human cell lines and a rat hepatoma cell line, as well as the h Ah R with native conformations. We concluded that the newly generated m Abs could be employed in AhR-based bioassays for analysis of environmental contaminants, and held great potential for further revealing the spatial structure of AhR and its biological functions in future studies.
