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J Neurosci. 2004 Sep 1;24(35):7663-73.

An element in the alpha1-tubulin promoter is necessary for retinal expression during optic nerve regeneration but not after eye injury in the adult zebrafish.

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  • 1University of Michigan, Mental Health Research Institute, Department of Biological Chemistry, Ann Arbor, Michigan 48109-0720, USA.

Abstract

We have shown previously that a 1.696 kb upstream fragment of the goldfish alpha1-tubulin promoter was capable of driving green fluorescent protein (GFP) expression in the developing and regenerating zebrafish CNS in a pattern closely mimicking the endogenous alpha1-tubulin gene. Comparison of fish and rat alpha1-tubulin promoters identified a 64 bp region with a conserved repetitive homeodomain (HD) consensus sequence core (TAAT) and a nearby basic helix-loop-helix binding E-box sequence (CANNTG), which led us to speculate that it could be of importance for regulating alpha1-tubulin gene transcription. To address this issue, we examined the ability of deletion mutants of the 1.696 kb promoter to drive expression of GFP in zebrafish retinal cells under normal conditions and after injury. Interestingly, although wild-type 1.696 kb and mutant promoters, lacking the E-box and/or HD sequences, exhibited rather similar patterns of GFP expression in the developing retina, significant differences were noticed in the mature retina. First, although the 1.696 kb promoter directed transgene expression to retinal neurons and progenitor cells, the activity of mutant promoters was drastically reduced. Second, we found that the E-box and HD sequences were necessary for transgene reinduction during optic nerve regeneration, but were not as important for transgene expression in regenerating retinal neurons after eye injury. In this latter lesion model, remarkably, both 1.696 kb and mutant promoters targeted GFP expression to Müller glia-like cells, some of which re-entered the cell cycle. These new findings will be useful for identifying the molecular signals necessary for successful CNS regeneration.

PMID:
15342733
DOI:
10.1523/JNEUROSCI.2281-04.2004
[PubMed - indexed for MEDLINE]
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