斑马鱼nr1d4a和nr1d4b基因的表达及其对不同环境刺激的响应-水生生物学专业论文.docxVIP

II II Abstract Members of the nuclear receptor family function as transcriptional factors and are involved in the regulation of multiple important biological processes. Previous studies have revealed that both nr1d4a and nr1d4b of zebrafish are immediate early genes that can be significantly induced by cold stress. However, the expression patterns of these genes and their transcriptional responses to other environmental stresses remain to be characterized. In the study, we cloned the cDNAs of zebrafish nr1d4a and nr1d4b, performed sequence alignment and phynogenetic analyses, and characterized the expression patterns and their transcriptional responses to several environmental stresses using real time quantitative RT-PCR (qPCR). The results indicate that zebrafish nr1d4a and nr1d4b genes are paralogs generated by genomic duplication. The DNA binding domain and ligand binding domain of these two paralogs are highly conserved, but these genes possess distinct expression patterns. The abundance of nr1d4a mRNA was quite low during the early stages of embryogenesis and markedly increased at 72hpf; however, nr1d4b was found to be maternally expressed and the mRNA abundance decreased at 6hpf and significantly increased at 72hpf as well. As for tissue-specific expression, the highest expression of zebrafish nr1d4a was found in brain and kidney, followed by gill, ovary, testis and eye, and liver is the organ with the lowest expression. nr1d4b mRNA is the most abundant in ovary, followed by testis and brain, and the lowest expression was found in intestine and heart. The expression of both nr1d4a and nr1d4b can be induced by multiple environmental stresses. The up-regulation of zebrafish nr1d4a and nr1d4b could be detected at as early as 0.5h after exposed to cold stress (16℃). However, the inductive effect of cold stress begins to decrease and gradually disappear after 6h of exposure. In addition to cold stress, the expression of zebrafish nr1d4a and nr1d4b was also ind