Int. J. Dev. Biol. 54: 731 - 736 (2010)
doi: 10.1387/ijdb.092969ls
© UPV/EHU Press

Characterization of an lhx1a transgenic reporter in zebrafish

Lisa M. Swanhart1, Nobuhiro Takahashi2, Rachel L. Jackson1, Gregory A. Gibson3, Simon C. Watkins3, Igor B. Dawid2 and Neil A. Hukriede*,1,4

1Department of Microbiology and Molecular Genetics, University of Pittsburgh, Pittsburgh, Pennsylvania, 2Laboratory of Molecular Genetics, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland, 3Department of Cell Biology and Physiology, Center for Biological Imaging, University of Pittsburgh, Pittsburgh, Pennsylvania, 4University of Pittsburgh Drug Discovery Institute, University of Pittsburgh, Pittsburgh, PA, USA.

ABSTRACT The LIM-domain containing transcription factor, Lhx1, is involved in the regulation of early gastrulation cell movements, kidney organogenesis and other processes in vertebrate model organisms. To follow the expression of this gene in live embryos, we created transgenic zebrafish expressing enhanced green fluorescent protein (EGFP) under the control of lhx1a regulatory regions. Tg(lhx1a:EGFP) pt303 recapitulates the expression of endogenous lhx1a beginning at early gastrula stages through 72 hours of development with only few exceptions. In addition, over-expression of the Nodal ligand, ndr1, results in the concomitant expansion of the transgene and endogenous lhx1a expression. Treatment of Tg(lhx1a:EGFP) pt303 embryos with the small molecule SB-431542, an inhibitor of Nodal signaling, results in the loss of both transgene and endogenous lhx1a expression. These experiments suggest that Tg(lhx1a:EGFP) pt303 is regulated in a manner similar to endogenous lhx1a. Therefore, this reporter can be utilized not only for monitoring lhx1a expression, but also for numerous applications, including chemical genetics screening.

Keywords:

lhx1a, transgene, zebrafish, kidney, gastrulation

*Corresponding author e-mail: hukriede@pitt.edu