Development of synapses between chick retinal neurons in dispersed culture

Morphological criteria allow several kinds of synapse to be recognized in the vertebrate retina. It is, however, not presently known if, or how, these morphological differences reflect physiological distinctions. Since a proper investigation of synaptic physiology in the intact retina is compromised by technical difficulties, we have examined dispersed cultures to discover if they are likely to provide a more tractable physiological preparation. The chief question addressed here concerns the extent to which normal synaptic development takes place in the impoverished conditions of dispersed cell culture. Cultures were established from embryonic day 8 chick retina and fixed for microscopy on embryonic equivalent (E.E.) days 12, 14, 16, and 18. Neuronal processes appeared shortly after plating and continued to increase in number and extent through E.E. 16. Cone cells were recognizable by virtue of their distinctive oil droplets. Two classes of cone could be distinguished on the basis of the density of their cytoplasmic staining. Presynaptic ribbons could be observed in cone cells on E.E. 12, but characteristic dyad and triad postsynaptic organization was seldom present at this stage nor was it often observed at subsequent times. An increase in the number of ribbon synapses in culture was seen on E.E. 18. These synapses may represent those of bipolar cells. Conventional synapses were found at all times examined but the number of these increased greatly between E.E. 14 and 16. Of these conventional synapses, we found some whose anatomy was characteristic of synapses made by amacrine cells as well as some whose anatomy was characteristic of synapses made by bipolar cells.