GENERATION OF CELL-LINES FROM EMBRYONIC QUAIL RETINA CAPABLE OF MATURE NEURONAL DIFFERENTIATION

The avian embryonic retina is widely used as a model system for cellul ar and molecular studies on central nervous system neurons. We aimed a t the generation of cell lines from the early embryonic quail retina b y retroviral oncogene transduction. For this, we made use of the retin a organ culture system which exhibits both proliferation, necessary fo r stable oncogene transduction, and initial neuronal differentiation, a prerequisite for the generation of cell lines with mature neuronal p roperties. The oncogene myc was chosen as it is both proliferation-ind ucing and differentiation-compatible. A chimeric gene., mycER, contain ing v-myc and the hormone-binding domain of the estrogen receptor, was used for transduction in order to allow for hormone regulation of myc activity. Transduced organ-cultured cells from temporal and nasal ret ina were passaged into sparse single cell cultures. From these, coloni es of rapidly dividing cells were isolated and the progeny expanded as cell lines. The lines contained cells with features of neuroepithelia l cells, showing vimentin and A2B5. They also contained spontaneously differentiated neuronal cells showing neurofilament L and N-CAM180. A subpopulation of the neuronal cells exhibited the morphological charac teristics of retinal ganglion cells, i.e., large pear-shaped somata ea ch emitting one long process with a distinct growth cone. In addition, they showed the marker profile of retinal ganglion cells, i.e., expre ssion of Thy-1, G4, DM-GRASP, Nr-CAM, neurofilament H, and tau. Neuron al differentiation could be induced by the addition of db cAMP and ret inoic acid. The mature neuronal features of the lines open new possibi lities to study properties of retinal neurons, including ganglion cell s, in a defined and manipulable experimental system. (C) 1995 Wiley-Li ss, Inc.