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Series GSE5338 Query DataSets for GSE5338
Status Public on Aug 18, 2006
Title In vivo function of NR2E3 in establishing photoreceptor identity during mammalian retinal development
Organism Mus musculus
Experiment type Expression profiling by array
Summary Rod and cone photoreceptors in mammalian retina are generated from common pool(s) of neuroepithelial progenitors. NRL, CRX and NR2E3 are key transcriptional regulators that control photoreceptor differentiation. Mutations in NR2E3, a rod-specific orphan nuclear receptor, lead to loss of rods, increased density of S-cones, and supernormal S-cone-mediated vision in humans. To better understand its in vivo function, NR2E3 was expressed ectopically in the Nrl-/- retina, where post-mitotic precursors fated to be rods develop into functional S-cones similar to the human NR2E3 disease. Expression of NR2E3 in the Nrl-/- retina completely suppressed cone differentiation and resulted in morphologically rod-like photoreceptors, which were not functional. Gene profiling of FACS-purified photoreceptors confirmed the role of NR2E3 as a strong suppressor of cone genes and an activator of a subset of rod genes (including rhodopsin) in vivo. Ectopic expression of NR2E3 in cone precursors and differentiating S-cones of wild type retina also generates rod-like cells. The dual regulatory function of NR2E3 is not dependent upon the presence of NRL and/or CRX, but on the timing and level of its expression. Our studies reveal a critical role of NR2E3 in establishing functional specificity of post-mitotic photoreceptor precursors during retinal neurogenesis.
Keywords: genetic modification
 
Overall design We mated the Crx::Nr2e3/Nrlko mice with the Nrl::GFP transgenic mice, in which the expression of GFP is driven by an Nrl promoter. Mouse retinas were dissected at 4 wk. GFP+ photoreceptors were enriched by FACS (FACSAria, BD Biosciences, Franklin Lakes, NJ). RNA was extracted from 1~5x105 flow-sorted cells using Trizol (Invitrogen). Total RNA (40-60 ng) was used for linear amplification with Ovation Biotin labeling system (Nugen), and 2.75 ug of biotin-labeled fragmented cDNA was hybridized to mouse GeneChips MOE430.2.0 (Affymetrix) having 45,101 probesets (corresponding to over 39,000 transcripts, and 34,000 annotated mouse genes).
Four independent samples were used at 4 weeks. We normalized
Crx::Nr2e3/Nrl-ko-Gfp data along with Nrl-ko-Gfp 4 weeks samples ( 4 replicates, refer to Series submission GSE4051). The normalized data was then subjected to two stage analysis based on False Discovery Rate Confidence Interval (FDR-CI) for screening differentially expressed genes (24, 27) with a minimum fold change of 4.
 
Contributor(s) Cheng H, Khanna R, Swaroop A
Citation(s) 16868010
Submission date Jul 18, 2006
Last update date Feb 11, 2019
Contact name Swaroop Anand
E-mail(s) swaroop@umich.edu
Phone 734-615 2246
URL http://www.umich.edu/~retina
Organization name University of Michigan
Department Ophthalmology & Visual Sciences
Street address 1000 Wall St.
City Ann Arbor
State/province MI
ZIP/Postal code 48105
Country USA
 
Platforms (1)
GPL1261 [Mouse430_2] Affymetrix Mouse Genome 430 2.0 Array
Samples (8)
GSM92610 Nrl-ko-Gfp 4 weeks retina replicate 1
GSM92611 Nrl-ko-Gfp 4 weeks replicate 2
GSM92612 Nrl-ko-Gfp 4 weeks replicate 3
Relations
BioProject PRJNA96359

Download family Format
SOFT formatted family file(s) SOFTHelp
MINiML formatted family file(s) MINiMLHelp
Series Matrix File(s) TXTHelp

Supplementary file Size Download File type/resource
GSE5338_RAW.tar 51.8 Mb (http)(custom) TAR (of CEL)
Raw data provided as supplementary file

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