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Purpose: Rod-cone gap junctions permit transmittal of rod visual information to the cone pathway. A recent report has shown that this transfer does not occur in mice in which the gap junction protein connexin 36 is knocked out indicating that rod-cone gap junctions are assembled from this protein. It remains unresolved, however, whether rods, cones or both express connexin 36. We have tried to address this question with the use of transgenic rod-less and cone-less mice., Methods: Deletion of Nrl, a transcription factor, results in a complete loss of rods with a concomitant increase in S-cones. We used this as the rod-less (cone-only) model. Cone-less (rod-only) retinas were from mice expressing an attenuated diphtheria toxin gene under the control of a promoter selective for cones. Nearly all long wavelength cones and 95% of short wavelength cones are missing in this model. Fixed retinal sections from these two models and age matched controls were used to detect connexin 36 gap junctions by immunofluorescence., Results: Punctate immunofluorescence, indicating the presence of gap junctions, was observed in the inner and outer plexiform layers of both wild type and cone-less and rod-less retinas. Our assumption was that immunofluorescence due to photoreceptor gap junctions would be observed in the outer plexiform layer. In all the animals, most of the immunofluorescence was in the inner plexiform layer, with only a marginal reaction in the outer plexiform layer. In cone-only (rod-less) retina, immunofluorescence in the outer plexiform layer increased by more than 20 fold compared to wild type. In rod-only (cone-less) retina, the outer plexiform layer showed about a 30% decrease in immunofluorescence. In both rod-less and cone-less retinas, immunofluorescence in the inner plexiform layer was higher than in the wild type by 25-50%., Conclusions: Cones constitute only about 3% of photoreceptors in the wild type retina while they make up 100% of the photoreceptors in cone-only retina. This increase in their numbers coincided with a 20 fold increase in immunofluorescence in the outer plexiform layer, strongly suggesting that cones express connexin 36. Conversely, when the cone numbers went down from 3% to near zero in cone-less retina, immunofluorescence decreased by about 30% in the outer plexiform layer, suggesting again that cones express the connexin and that they contribute to its presence disproportionately more than their numbers indicate. The results from both rod-less and cone-less animals are strongly indicative of cones expressing connexin 36, but are not sufficient to conclude whether rods express the protein. An unexpected observation from our experiments is that immunofluorescence increases slightly in the inner plexiform layer in both rod-less and cone-less retina for reasons that need further investigation.

We have examined the role of neighbor relationships between cholinergic amacrine cells upon their positioning and dendritic field size by producing partial ablations of this population of cells during early development. We first determined the effectiveness of L-glutamate as an excitotoxin for ablating cholinergic amacrine cells in the developing mouse retina. Subcutaneous injections (4 mg/g) made on P-3 and thereafter were found to produce a near-complete elimination, while injections at P-2 were ineffective. Lower doses on P-3 produced only partial reductions, and were subsequently used to examine the effect of partial ablation upon mosaic organization and dendritic growth of the remaining cells. Four different Voronoi-based measures of mosaic geometry were examined in L-glutamate-treated and normal (saline-treated) retinas. Partial depletions of around 40% produced cholinergic mosaics that, when scaled for density, approximated the mosaic geometry of the normal retina. Separate comparisons simulating a 40% random deletion of the normal retina produced mosaics that were no different from those experimentally depleted retinas. Consequently, no evidence was found for positional regulation in the absence of normal neighbor relationships. Single cells in the ganglion cell layer were intracellularly filled with Lucifer Yellow to examine the morphology and dendritic field extent following partial ablation of the cholinergic amacrine cells. No discernable effect was found on their starburst morphology, and total dendritic field area, number of primary dendrites, and branch frequency were not significantly different. Cholinergic amacrine cells normally increase their dendritic field area after P-3 in excess of retinal expansion; despite this, the present results show that this growth is not controlled by the density of neighboring processes.

During development, retinal ganglion cells undergo conspicuous structural remodeling as they gradually attain their mature morphology and connectivity. Alterations in their dendritic organization and in their axonal projections can also be achieved following early insult to their targets or their afferents. Other retinal cell types are thought not to display this same degree of developmental plasticity. The present review will consider the evidence, drawn largely from recent experimental studies in the carnivore retina, that photoreceptors also undergo structural remodeling, extending their terminals transiently into inner plexiform layer before retracting to the outer plexiform layer. The determinants of this transient targeting to the inner plexiform layer are considered, and the role of cholinergic amacrine cells is discussed. The factors triggering this retraction are also considered, including the concurrent maturational changes in outer segment formation and in the differentiation of the outer plexiform layer. These results provide new insight into the life history of the photoreceptor cell and its connectivity, and suggest a transient role for the photoreceptors in the circuitry of the inner retina during early development, prior to the onset of phototransduction.