Investigative Ophthalmology & Visual Science · 2005 · 27 citations · 24 references
Life SciencesGeneticsNeuroscienceNeurogeneticsRecombinant Dna TechnologyRecombinationRecombination ProteinsRetinal DegenerationRetinal DiseaseRetinal DisordersRetinal NeurobiologyRetinal NeuroscienceBacterial Artificial Chromosome TransgenesisBacterial Artificial Chromosome VectorsCerebellar Purkinje CellsInner Nuclear LayerPurkinje Cell LayerTransgenic Cell Line EngineeringTransgenic MiceTransgenic Mouse Models
PURPOSE: To establish a transgenic mouse line that expresses Cre-recombinase in retinal rod bipolar cells for the generation of rod bipolar cell-specific knockout mutants. METHODS: The IRES-Cre-cDNA fragment was inserted into a 173-kb bacterial artificial chromosome (BAC) carrying the intact Pcp2 gene, by using red-mediated recombineering. Transgenic mice were generated with the modified BAC and identified. The Cre-transgenic mice were crossed with ROSA26 and Z/EG reporter mice to detect Cre-recombinase activity. RESULTS: X-gal staining showed that strong Cre-recombinase activities were present in retinal inner nuclear layers and cerebellar Purkinje cells. Double staining with an anti-GFP antibody and an anti-PKCalpha antibody (specific for retinal rod bipolar cells) revealed that Cre-recombinase activity localized exclusively to the rod bipolar cells in the retina. CONCLUSIONS: A mouse BAC-Pcp2-IRES-Cre transgenic line that expresses Cre-recombinase in retinal rod bipolar neurons has been established. Because mutations in some ubiquitously expressed genes may result in retinal degenerative diseases, the mouse strain BAC-Pcp2-IRES-Cre will be a useful new tool for investigating the effects of retinal rod bipolar cell-specific gene inactivation.
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Cell differentiation in the retina of the mouse
Richard W. Young · The Anatomical Record · 1985 · 1.1K citations