Proximal upstream region of zebrafish bone morphogenetic protein 4 promoter directs heart expression of green fluorescent protein.
We examined the activity of the bone morphogenetic protein 4 (BMP4) promoter in zebrafish embryos via transient and stable transgenic expression analyses in order to obtain a better understanding of the regulation of BMP4 tissue-specific expression. Transient expression studies showed that the 9.0-kb BMP4 promoter/upstream region drove green fluorescent protein (GFP) expression mainly in the heart. Deletion analyses indicated the existence of multiple regulatory elements in the 7.5-kb BMP4 promoter/proximal upstream region. In addition, a coinjection experiment further demonstrated the 2.4-kb Bgl II-Hind III DNA region contains major positive regulatory elements. In addition, stable transgenic lines were established to further confirm the heart-specificity of this segment in BMP4 promoter. The results showed that GFP was mainly localized in the myocardium of developing ventricles of 48-hpf (hours postfertilization), 72-hpf, and 100-hpf transgenic F(1) embryos. Together, these results indicate that the 7.5-kb BMP4 promoter/proximal upstream region specifically contains regulatory elements for BMP4 expression in the heart, while regulatory elements for other endogenous BMP4-expressing tissues may reside in more distal regions and/or in introns.
Shentu
Hsuan
H
Institute of Zoology, Academia Sinica, Nankang, Taipei, Taiwan, ROC.
Wen
Hui-Ju
HJ
Her
Guor Mour
GM
Huang
Chang-Jen
CJ
Wu
Jen-Leih
JL
Hwang
Sheng-Ping L
SP
D000818:Animals; D055415:Bone Morphogenetic Protein 4; D019485:Bone Morphogenetic Proteins / Q000235:genetics*; D047108:Embryonic Development; D018507:Gene Expression Regulation, Developmental; D049452:Green Fluorescent Proteins; D006321:Heart / Q000502:physiology; D007202:Indicators and Reagents / Q000032:analysis; D008164:Luminescent Proteins / Q000032:analysis / Q000096:biosynthesis*; D011401:Promoter Regions, Genetic; D015027:Zebrafish / Q000196:embryology / Q000235:genetics*; D029961:Zebrafish Proteins
D055415:Bone Morphogenetic Protein 4; D019485:Bone Morphogenetic Proteins; D007202:Indicators and Reagents; D008164:Luminescent Proteins; D029961:Zebrafish Proteins; C525813:bmp4 protein, zebrafish; D049452:Green Fluorescent Proteins