Targeting of GFP to Newborn Rods by Nrl Promoter and Temporal Expression Profiling of Flow-Sorted Photoreceptors

The Maf-family transcription factor Nrl is a key regulator of photoreceptor differentiation in mammals. Ablation of the Nrl gene in mice leads to functional cones at the expense of rods. We show that a 2.5-kb Nrl promoter segment directs the expression of enhanced GFP specifically to rod photorecept...

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Published inProceedings of the National Academy of Sciences - PNAS Vol. 103; no. 10; pp. 3890 - 3895
Main Authors Akimoto, Masayuki, Cheng, Hong, Zhu, Dongxiao, Brzezinski, Joseph A., Khanna, Ritu, Filippova, Elena, Oh, Edwin C. T., Jing, Yuezhou, Linares, Jose-Luis, Brooks, Matthew, Zareparsi, Sepideh, Mears, Alan J., Hero, Alfred, Glaser, Tom, Swaroop, Anand
Format Journal Article
LanguageEnglish
Published United States National Academy of Sciences 07.03.2006
National Acad Sciences
SeriesFrom the Cover
Subjects
Animals
Animals, Newborn
Antibodies
Basic-Leucine Zipper Transcription Factors - deficiency
Basic-Leucine Zipper Transcription Factors - genetics
Basic-Leucine Zipper Transcription Factors - metabolism
Biological Sciences
Cell cycle
Cell Differentiation
Developmental stages
DNA-Binding Proteins - genetics
Endocrine system
Eye Proteins - genetics
Eye Proteins - metabolism
Gene expression
Gene Expression Profiling
Genes
Green Fluorescent Proteins - genetics
Green Fluorescent Proteins - metabolism
Humans
Mice
Mice, Knockout
Mice, Transgenic
Neurons
Neurons - cytology
Neurons - metabolism
Neurosciences
Photoreceptor Cells, Vertebrate - cytology
Photoreceptor Cells, Vertebrate - metabolism
Photoreceptors
Promoter Regions, Genetic
Recombinant Proteins - genetics
Recombinant Proteins - metabolism
Retina
Retina - cytology
Retina - growth & development
Retina - metabolism
Retinal Cone Photoreceptor Cells - cytology
Retinal Cone Photoreceptor Cells - growth & development
Retinal Cone Photoreceptor Cells - metabolism
Retinal Rod Photoreceptor Cells - cytology
Retinal Rod Photoreceptor Cells - growth & development
Retinal Rod Photoreceptor Cells - metabolism
Retinal rods
Rodents
Transgenic animals
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Summary:The Maf-family transcription factor Nrl is a key regulator of photoreceptor differentiation in mammals. Ablation of the Nrl gene in mice leads to functional cones at the expense of rods. We show that a 2.5-kb Nrl promoter segment directs the expression of enhanced GFP specifically to rod photoreceptors and the pineal gland of transgenic mice. GFP is detected shortly after terminal cell division, corresponding to the timing of rod genesis revealed by birthdating studies. In$Nrl^{-/-}$retinas, the GFP+ photoreceptors express S-opsin, consistent with the transformation of rod precursors into cones. We report the gene profiles of freshly isolated flow-sorted GFP+ photoreceptors from wild-type and$Nrl^{-/-}$retinas at five distinct developmental stages. Our results provide a framework for establishing gene regulatory networks that lead to mature functional photoreceptors from postmitotic precursors. Differentially expressed rod and cone genes are excellent candidates for retinopathies.
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Edited by Jeremy Nathans, Johns Hopkins University School of Medicine, Baltimore, MD, and approved January 6, 2006
Author contributions: M.A., H.C., A.J.M., and A.S. designed research; M.A., H.C., J.A.B., E.F., E.C.T.O., and M.B. performed research; M.A., H.C., D.Z., R.K., E.C.T.O., Y.J., J.-L.L., and A.H. contributed new reagents/analytic tools; M.A., H.C., D.Z., J.A.B., R.K., Y.J., S.Z., T.G., and A.S. analyzed data; and D.Z., A.H., T.G., and A.S. wrote the paper.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.0508214103