Characterization Of Pou4f2/Brn3b-GFP Knock-in Mouse Line

Eyes are the organ received different Optical signals of the outside world inmammal. The mammalian retina consists of neurons of more than60distinct subtypes.These different cell types can be grouped into six major neuronal cell types and oneglial type. Mammalian eye development has been an excellent model system toinvestigate CDS, due to clarity structure of the retina, multiple cell types and clearphysiological function. Prior studies have reported POU-IV family transcription factorPou4f2(Brn3b) express in the initial stage of retina development in mouse.Pou4f2-null mutation leads a loss of70%of retinal ganglion cells. Deletion of Pou4f2results in RGC axon defect and apoptosis. In order to find the downstream genes ofPou4f2, we report the generation of knock-in Pou4f2-GFP mouse which we showreliably express Pou4f2-GFP fusion protein. The Pou4f2-GFP targeting construct wasgenerated by inserting GFP coding sequence that end upstream of the translationalinitiation codon ATG. The specific expression of Pou4f2-GFP fusion protein in thedeveloping retina was revealed through IHC. We also detected the expression of Isl1and Pou4f1in mutant retina is same as in wild type. Both anti-GFP and anti-Pou4f2antibodies co-precipitated with the promoters of Shh and Isl2. Pou4f2-GFP expressionin the retina of Pou4f2-GFP mouse embryos can be directly observed by confocalmicroscope. The results showed Pou4f2-GFP mouse can be used to the model ofinduced retinal ganglion cells differentiation.