![](/coreweb/template1/pix/main_left_bg.gif) |
![](/coreweb/template1/pix/pixel.gif) |
GEO help: Mouse over screen elements for information. |
|
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 |
Supplementary file |
Size |
Download |
File type/resource |
GSE5338_RAW.tar |
51.8 Mb |
(http)(custom) |
TAR (of CEL) |
|
|
|
|
![](/coreweb/template1/pix/main_right_bg.gif) |