Toxicity of TCDD in Zebrafish Embryonic Vascularization and Skeletogenesis

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基本資料

Project title

Code/計畫編號

NSC88-2313-B019-042

Translated Name/計畫中文名

戴奧辛對斑馬魚胚胎發育時期血管與骨骼生成之生理毒性研究

Funding Organization/主管機關

National Science and Technology Council

Department/Unit

Department of Bioscience and Biotechnology

Website

https://www.grb.gov.tw/search/planDetail?id=434431

Year

1999

Start date/計畫起

01-08-1998

Expected Completion/計畫迄

01-07-1999

Bugetid/研究經費

755千元

ResearchField/研究領域

生物科學
漁業

ARNT是一群具有相當保守性的basic-helix-loop-helix結構的轉錄蛋白,在各種生物體中扮演著調節生理機能與胚胎發育的重要角色。先前我們利用RT-PCR的方式成功的從斑馬魚胚胎與組織中獲得一段類似哺乳動物ARNT2蛋白的cDNA序列。為了進一步瞭解此一魚類的ARNT2蛋白的生物功能,我們成功的自斑馬魚胚胎基因庫中篩選到一完整的cDNA序列,我們稱為zARNT2A,其中包括了完整的basic-helix-loop-helix區域與PAS A/B區域,但缺少了C-terminal的活化功能區。在培養的細胞株內,外生性的zARNT2A表現會明顯抑制戴奧辛所活化的CYP1A基因表現。這顯示此一蛋白為抑制性基因調控子。利用全覆式RNA原位雜交的方法,我們觀察到zARNT2A在胚胎發育的前期有分布全身的表現,進而隨著發育的進展,zARNT2A mRNA的分布逐漸集中在腦部與原腎區域。在成魚的組織中,我們也可以利用RT-PCR的方式偵測到zARNT2A廣泛的在許多組織中表現,顯示zARNT2A在胚胎發育與其他重要生理機能中扮演相當重要的角色。[本論文為專題計畫之部分成果,已獲得Marine Biotechnology初審有條件通過,目前正依據審查意見修正中,並將於近日完成。其他研究成果亦將整理後投稿於SCI期刊發表] Aryl hydrocarbon receptor nuclear translocator (ARNT) factors belong to a novel basic-helix-loop-helix-PAS (bHLH-PAS) transcription factor family which controls a variety of physiological and developmental processes. In previous study, we have obtained a partial fragment of ARNT2-like factor cDNA from zebrafish embryo, liver and other tissue by RT-PCR. In an effort to characterize the function of this factor, we screened an embryonic cDNA library and obtained the cDNA of this ARNT2-like factor, zARNT2A. The deduced protein sequence encompasses the basic-helix-loop-helix and PAS-A/B motifs and shares highest sequence similarity with the N-terminal half of mouse ARNT2 factor. However, it lacks a C-terminal transactivation motif following the PAS-A/B motifs. Transient expression of zARNT2A in cultured cells resulted in repression of TCDD-dependent CYP1A transcription. Whole-mount in situ hybridization revealed that zARNT2A is expressed in brain and pronephros at prime-5 stages. In adult fish, zARNT2A mRNA is transcribed in a wide range of tissue, which indicates that zARNT2A and its corresponding signal transduction mechanisms have important roles in fish development and other physiological aspects.

Keyword(s)

戴奧辛
細胞毒性
血管生成
骨骼生成
基因表現
斑馬魚
Dioxin
Cytotoxicity
Vascularization
Skeletogenesis
Gene expression
Zebrafish

DSpace CRIS

Toxicity of TCDD in Zebrafish Embryonic Vascularization and Skeletogenesis

基本資料

Description