Comment[ArrayExpressAccession]	E-GEOD-45835
MAGE-TAB Version	1.1						
Public Release Date	2013-04-06
Investigation Title	Expression profiling of Ascl1-reprogrammed P12 Müller glia compared with freshly dissociated progenitors and Müller glia						
Comment[Submitted Name]	Expression profiling of Ascl1-reprogrammed P12 Müller glia compared with freshly dissociated progenitors and Müller glia
Experiment Description	Non-mammalian vertebrates have a robust ability to regenerate injured retinal neurons from Müller glia cells (MG) that activate the proneural factor Achaete-scute homolog 1 (Ascl1/Mash1) and de-differentiate into progenitors cells. In contrast, mammalian MG have a limited regenerative response and fail to upregulate Ascl1 after injury. To test whether Ascl1 could restore a neurogenic potential to mammalian MG, we over-expressed Ascl1 in dissociated mouse MG cultures and intact retinal explants. Ascl1-infected MG upregulate retinal progenitor-specific genes, while downregulating glial genes. Furthermore, Ascl1 remodeled the chromatin at its targets from a repressive to active configuration. MG-derived progenitors differentiated into cells that exhibited neuronal morphologies, expressed retinal subtype-specific neuronal markers, and displayed neuron-like physiological responses. These results indicate that a single transcription factor, Ascl1, can produce a neurogenic state in mature Muller glia. Expresssion profiling was used to determine the genes that were changed after Ascl1 infection of P12 cultured Müller glia compared with those present in P0 progenitors and P7-P21 Müller glia Retinas were dissociated and FAC-sorted from Hes5-GFP mice at P0, P7, P10, P14 or P21 and submitted for profiling. WT Retinas were dissociated at P12, grown for 1 week in culture, and infected with lentiviruses expressing Ascl1 or GFP for four days. Total RNA was extracted and submitted for profiling.						
Term Source Name	ArrayExpress	EFO
Term Source File	http://www.ebi.ac.uk/arrayexpress/	http://www.ebi.ac.uk/efo/efo.owl					
Person Last Name	REH	Pollak	Reh				
Person First Name	THOMAS	Julia	Thomas				
Person Mid Initials	A		A				
Person Email	tomreh@uw.edu						
Person Affiliation	UNIVERSITY OF WASHINGTON						
Person Address	BIOLOGICAL STRUCTURE, UNIVERSITY OF WASHINGTON, 1959 NE PACIFIC STREET, SEATTLE, WA, USA						
Person Roles	submitter						
Protocol Name	P-GSE45835-1	P-GSE45835-5	P-GSE45835-6	P-GSE45835-2	P-GSE45835-3	P-GSE45835-4	P-GSE45835-7
Protocol Description	The data were analyzed with Affymetrix Expression Console Software ID_REF =  VALUE = log2 RMA signal	Cy3 RNA were prepared according to the standard Affymetrix protocol from total RNA (Expression Analysis Technical Manual, 2001, Affymetrix).	Following fragmentation, cRNA were hybridized on Mouse Gene 1.0 ST Array, washed, and stained as per manufacturer guidelines.	N/A	N/A	Total RNA was extracted using Picopure RNA Isolation kit (Invitrogen, per manufacturer's instructions) or TRIzol (manufacturer's instructions) and followed with RNA cleaup kit (Qiagen)	Genechips were scanned using Affymetrix GeneChip Scanner 3000
Protocol Type	normalization data transformation protocol	labelling protocol	hybridization protocol	sample treatment protocol	growth protocol	nucleic acid extraction protocol	array scanning protocol
Experimental Factor Name	CELL TYPE	DAYS POST-INFECTION	GENOTYPE	AGE	CONDITION		
Experimental Factor Type	cell type	days post-infection	genotype	age	condition		
Comment[SecondaryAccession]	GSE45835						
Comment[GEOReleaseDate]	2013-04-06						
Comment[ArrayExpressSubmissionDate]	2013-04-06						
Comment[GEOLastUpdateDate]	2013-04-08						
Comment[AEExperimentType]	transcription profiling by array						
SDRF File	E-GEOD-45835.sdrf.txt