Figure 9. Transfection of E10 chick retinal cultures with truncated IRBP promoter (-70/+101 fragment) driving the enhanced green fluorescent protein (EGFP) gene.

Primary cultures of E10 chick retina cells were transiently transfected via calcium phosphate precipitation with 10 µg of the plasmid p70-EGFP (a HindIII/BamHI -70 to +101 fragment of the mouse IRBP promoter cloned into pEGFP-1). Photos were taken three days after transfection.

(a) Photographs are of three representative cells that fluoresce after transfection of the cultures. Morphology suggests an axon with pedicle at the bottom, a cell body in the middle, and an inner segment with a rudimentary outer segment near the top of the photographs.

    

(b) Comparison of fluorescence and bright field exposure of putative photoreceptor transfected with p70-EGFP. The top photograph is with fluorescence lighting only. The bottom photograph is the same field with the transmission light turned up enough to observe an oil droplet in the same cell. The presence of an oil droplet is characteristic of photoreceptor cells in chickens.

(c) Photograph of typical multipolar, neuron-like cell. Hundreds of multipolar cells with extensive arborization and lacking oil droplets fluoresced in retinal cultures transfected with p70-EGFP.