水生生物学报
水生生物學報
수생생물학보
ACTA HYDROBIOLOGICA SINICA
2014年
1期
100-107
,共8页
杨玲%严军军%龙勇%崔宗斌%鲍传和
楊玲%嚴軍軍%龍勇%崔宗斌%鮑傳和
양령%엄군군%룡용%최종빈%포전화
斑马鱼%细胞核受体%环境刺激%基因表达
斑馬魚%細胞覈受體%環境刺激%基因錶達
반마어%세포핵수체%배경자격%기인표체
Zebrafish%Nuclear receptor%Environmental stresses%Gene expression
研究获得了斑马鱼 nr1d4a 和 nr1d4b 基因的 cDNA,进行了序列比对和系统进化分析,并采用实时定量RT-PCR (qPCR)方法研究了其表达模式及对不同环境刺激的转录反应。研究发现,斑马鱼nr1d4a和nr1d4b是由基因复制产生的旁系同源基因,具有高度保守的DNA结合结构域和配体结合结构域。斑马鱼nr1d4a和nr1d4b的表达模式具有明显的差别。nr1d4a在胚胎发育早期的表达量很低,72 hpf时开始显著升高；而nr1d4b具有较高水平的母源性表达,6 hpf时的表达量明显降低,但也在72 hpf显著回升。nr1d4a在脑和肾脏中表达量最高,其次是鳃、卵巢、精巢和眼,在肝脏中的表达量最低； nr1d4b 在卵巢中表达量最高,其次是精巢和脑,在肠道和心脏中表达量最低。斑马鱼nr1d4a和nr1d4b都能被多种环境刺激瞬时诱导表达。16℃低温处理0.5h就能显著诱导斑马鱼nr1d4a和nr1d4b基因的表达,但处理6h后其诱导效应开始下降并逐渐消失。除低温外,重金属(2μmol/L镉)、缺氧(5%氧气)和盐度(5‰)处理均能瞬时诱导nr1d4a和nr1d4b的表达,说明 nr1d4a和nr1d4b基因可能参与斑马鱼对多种环境刺激的适应性反应。研究为深入揭示鱼类 nr1d4a和nr1d4b基因的生物学功能及其表达调控机制奠定了基础。
研究穫得瞭斑馬魚 nr1d4a 和 nr1d4b 基因的 cDNA,進行瞭序列比對和繫統進化分析,併採用實時定量RT-PCR (qPCR)方法研究瞭其錶達模式及對不同環境刺激的轉錄反應。研究髮現,斑馬魚nr1d4a和nr1d4b是由基因複製產生的徬繫同源基因,具有高度保守的DNA結閤結構域和配體結閤結構域。斑馬魚nr1d4a和nr1d4b的錶達模式具有明顯的差彆。nr1d4a在胚胎髮育早期的錶達量很低,72 hpf時開始顯著升高；而nr1d4b具有較高水平的母源性錶達,6 hpf時的錶達量明顯降低,但也在72 hpf顯著迴升。nr1d4a在腦和腎髒中錶達量最高,其次是鰓、卵巢、精巢和眼,在肝髒中的錶達量最低； nr1d4b 在卵巢中錶達量最高,其次是精巢和腦,在腸道和心髒中錶達量最低。斑馬魚nr1d4a和nr1d4b都能被多種環境刺激瞬時誘導錶達。16℃低溫處理0.5h就能顯著誘導斑馬魚nr1d4a和nr1d4b基因的錶達,但處理6h後其誘導效應開始下降併逐漸消失。除低溫外,重金屬(2μmol/L鎘)、缺氧(5%氧氣)和鹽度(5‰)處理均能瞬時誘導nr1d4a和nr1d4b的錶達,說明 nr1d4a和nr1d4b基因可能參與斑馬魚對多種環境刺激的適應性反應。研究為深入揭示魚類 nr1d4a和nr1d4b基因的生物學功能及其錶達調控機製奠定瞭基礎。
연구획득료반마어 nr1d4a 화 nr1d4b 기인적 cDNA,진행료서렬비대화계통진화분석,병채용실시정량RT-PCR (qPCR)방법연구료기표체모식급대불동배경자격적전록반응。연구발현,반마어nr1d4a화nr1d4b시유기인복제산생적방계동원기인,구유고도보수적DNA결합결구역화배체결합결구역。반마어nr1d4a화nr1d4b적표체모식구유명현적차별。nr1d4a재배태발육조기적표체량흔저,72 hpf시개시현저승고；이nr1d4b구유교고수평적모원성표체,6 hpf시적표체량명현강저,단야재72 hpf현저회승。nr1d4a재뇌화신장중표체량최고,기차시새、란소、정소화안,재간장중적표체량최저； nr1d4b 재란소중표체량최고,기차시정소화뇌,재장도화심장중표체량최저。반마어nr1d4a화nr1d4b도능피다충배경자격순시유도표체。16℃저온처리0.5h취능현저유도반마어nr1d4a화nr1d4b기인적표체,단처리6h후기유도효응개시하강병축점소실。제저온외,중금속(2μmol/L력)、결양(5%양기)화염도(5‰)처리균능순시유도nr1d4a화nr1d4b적표체,설명 nr1d4a화nr1d4b기인가능삼여반마어대다충배경자격적괄응성반응。연구위심입게시어류 nr1d4a화nr1d4b기인적생물학공능급기표체조공궤제전정료기출。
We cloned the cDNAs of zebrafishnr1d4a andnr1d4b, performed sequence alignment and phynogenetic analyses, and characterized the expression patterns and their transcriptional responses to several environmental stresses by using real time quantitative RT-PCR (qPCR). The results indicated that zebrafishnr1d4a andnr1d4b 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 ofnr1d4a mRNA was quite low during the early stages of embryogenesis and markedly increased at 72hpf; however,nr1d4b was found to be ma-ternally expressed and the mRNA abundance decreased at 6hpf and significantly increased at 72hpf as well. As for tis-sue-specific expression, the highest expression of zebrafishnr1d4a was found in brain and kidney, followed by gill, ovary, testis and eye, and liver was the organ with the lowest expression. The highest abundance ofnr1d4b mRNA was found in ovary, followed by testis and brain, and the lowest expression was found in intestine and heart. The expression of both nr1d4a andnr1d4b could be induced by multiple environmental stresses. The up-regulation of zebrafishnr1d4a andnr1d4b could be detected at as early as 0.5h after exposed to cold stress (16℃). However, the inductive effect of cold stress decreased and gradually disappeared after 6h of exposure. In addition to cold stress, the expression of ze-brafish nr1d4a andnr1d4b was also induced by heavy metal (2μmol/L cadmium), hypoxia (5% oxygen) and salinity (5‰), indicating the involvement of these genes in the acclimation to various environmental stresses. Thus, these find-ings have laid the foundation for further investigation ofnr1d4a andnr1d4b functions and mechanisms underlying the regulation of their expression in fish.
