Drs. Huang and Jou contributed equally to this work.
Conditional expression of a myocardium-specific transgene in zebrafish transgenic lines
Article first published online: 23 JUN 2005
Copyright © 2005 Wiley-Liss, Inc.
Volume 233, Issue 4, pages 1294–1303, August 2005
How to Cite
Huang, C.-J., Jou, T.-S., Ho, Y.-L., Lee, W.-H., Jeng, Y.-T., Hsieh, F.-J. and Tsai, H.-J. (2005), Conditional expression of a myocardium-specific transgene in zebrafish transgenic lines. Dev. Dyn., 233: 1294–1303. doi: 10.1002/dvdy.20485
- Issue published online: 15 JUL 2005
- Article first published online: 23 JUN 2005
- Manuscript Accepted: 4 MAR 2005
- Manuscript Revised: 15 FEB 2005
- Manuscript Received: 14 OCT 2004
- National Science Council of Taiwan. Grant Numbers: NSC93-2313-B002-058, NSC93-3112-B-002-32
- National Taiwan University Hospital. Grant Number: 92A-1002
- cardiac myosin light chain 2
To develop the first heart-specific tetracycline (Tet)-On system in zebrafish, we constructed plasmids in which the cardiac myosin light chain 2 promoter of zebrafish was used to drive the reverse Tet-controlled transactivator (rtTA) and the green fluorescent protein (GFP) reporter gene was preceded by an rtTA-responsive element. In the zebrafish fibroblast cell-line, rtTA-M2, one of rtTA's derivatives, demonstrated the highest increase in luciferase activity upon doxycycline (Dox) induction. We then generated two germ lines of transgenic zebrafish: line T03 was derived from microinjection of a plasmid containing rtTA-M2 and a plasmid containing a responsive reporter gene, whereas line T21 was derived from microinjection of a single dual plasmid. Results showed that line T21 was superior to line T03 in terms of greater GFP intensity after induction and with of minimal leakiness before induction. The photographic images of induced GFP in the heart of F2 larvae showed that the fluorescent level of GFP was dose-responsive. The level of GFP expressed in the F3 3 days postfertilization larvae that were treated with Dox for 1 hr decreased gradually after the withdrawal of the inducer; and the fluorescent signal disappeared after 5 days. The GFP induction and reduction were also tightly controlled by Dox in the F3 adult fish from line T21. This Tet-On system developed in zebrafish shows much promise for the study of the gene function in a specific tissue at the later developmental stage. Developmental Dynamics 233:1294–1303, 2005. © 2005 Wiley-Liss, Inc.