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LF & Too Molecular Brain Research 139 (2005) 1 – 12.pdf
LF & Too Molecular Brain Research 139 (2005) 1 – 12.pdf
Tissue expression of alternatively spliced GFRalpha1, NCAM and RET isoforms and the distinct functional consequence of ligand-induced activation of GFRalpha1 isoforms
Introduction
Materials and methods
Reverse transcription (RT) reaction
Sequence independent real-time PCR using SYBR Green I Plasmids construction
Sequence independent real-time PCR
Stable transfection of Neuro2A cells
Analysis of MAP Kinase (Erk/12) phosphorylation
Immunocytochemistry
Microarray analyses and verification with real-time PCR
Data analyses
Results
Development of a highly specific and sensitive quantitative real-time PCR assay
GFRalpha1b is expressed significantly in mouse tissues
RET9 is predominantly expressed in mouse tissues
NCAM120, 140 and 180 are expressed at equivalent levels in the brain
Neuro2A cells express NCAM and RET isoforms endogenously
NTN and GDNF activate ERK1/2 in GFRalpha1 isoform transfected cells
Profiles of ligand-induced gene expression in GFRalpha1 isoform transfected cells