Development of the cone photoreceptor mosaic in the mouse retina revealed by fluorescent cones in transgenic mice.

PURPOSE Normal function of the retina relies on the orderly stereotyped organization of different neurons and their synaptic connections. How such neural organization is patterned during development remains poorly understood due to the paucity of adequate developmental markers. This study was to examine the spatial organization and development of cone photoreceptors quantitatively in the mouse retina. METHODS A transgenic approach was used to generate a living cone cell marker by driving GFP expression in mouse cones with the human red/green opsin gene 5' sequences. The spatial organization and development of the cones in the mouse retinas were examined quantitatively with epifluorescence and scanning laser confocal microscopy. Cone specific GFP expression in the developing retinas was verified with peanut agglutinin (PNA) staining. Developmental expression of mouse cone opsin genes was determined with RT-PCR. RESULTS The fluorescent retinal cells expressing GFP can be visualized as early as on embryonic day E15. Following up morphological differentiation of these cells revealed features that were consistent with the typical morphology of the mouse cones. Double labeling with cone specific PNA showed that these cells were co-labeled starting from postnatal day P1, and that a subpopulation of PNA positive cones expressed the GFP. The fluorescent cell densities had a similar ventral and dorsal distribution from E15 to P2, increased dramatically in the ventral by P6, and in the dorsal from P7. Nearest neighbor distance analysis demonstrated that this subpopulation of cones was organized into a regular mosaic pattern with a regularity index of 4.82 in the central and 3.55 in the peripheral retina. Quantitative pattern assessment of the developing cones revealed that the fluorescent cells appeared to be distributed in a non-random array before birth. The regularity of the cone array began to rise on P7, in parallel with the onset of mouse green opsin gene expression and the development of cone pedicles. The regular pattern of cone mosaic organization was basically formed by P10, coinciding with the timing of the cone pedicle maturation. CONCLUSIONS The cones in the mouse retina are organized in a regular mosaic pattern. Patterning the cone mosaic appears to follow a two phase developmental process involving regulated opsin gene expression and cone pedicle maturation: an early phase where a non-random array emerges during cone differentiation, and a late phase where the regular mosaic pattern is mature at the time when cone synaptic contacts are being formed.