The Journal of Neuroscience, September 1, 2004, 24(35):7663-7673; doi:10.1523/JNEUROSCI.2281-04.2004
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Development/Plasticity/Repair
An Element in the
1-Tubulin Promoter Is Necessary for Retinal Expression during Optic Nerve Regeneration But Not after Eye Injury in the Adult Zebrafish
Marie-Claude Senut,
Abhilasha Gulati-Leekha, and
Daniel Goldman
University of Michigan, Mental Health Research Institute, Department of Biological Chemistry, Ann Arbor, Michigan 48109-0720
We have shown previously that a 1.696 kb upstream fragment of the goldfish
1-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
1-tubulin gene. Comparison of fish and rat
1-tubulin promoters identified a 64 bp region with a conserved repetitive homeodomain (HD) consensus sequence core (TAAT) and a nearby basic helixloop-helix binding E-box sequence (CANNTG), which led us to speculate that it could be of importance for regulating
1-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.
Key words: tubulin; gene expression; homeodomain; E-box; regeneration; retina; visual system; zebrafish
Received Jan 21, 2004;
revised July 12, 2004;
accepted July 14, 2004.
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