Neurogenesis in the retinal ganglion cell layer of the rat.

The present study has examined the birthdates of neurons in the retinal ganglion cell layer of the adult rat. Rat fetuses were exposed to tritiated thymidine in utero to label neurons departing the mitotic cycle at different gestational stages from embryonic days 12 through to 22. Upon reaching adulthood, rats were either given unilateral injections of horseradish peroxidase into target visual nuclei in order to discriminate (1) ganglion cells from displaced amacrine cells, (2) decussating from non-decussating ganglion cells, and (3) alpha cells from other ganglion cell types; or, their retinae were immunohistochemically processed to reveal the choline acetyltransferase-immunoreactive amacrine cells in the ganglion cell layer. Retinae were embedded flat in resin and cut en face to enable reconstruction of the distribution of labelled cells. Retinal sections were autoradiographically processed and then examined for neurons that were both tritium-positive and either horseradish peroxidase-positive or choline acetyltransferase-positive. Tritium-positive neurons in the ganglion cell layer were present in rats that had been exposed to tritiated thymidine on embryonic days E14-E22. Retinal ganglion cells were generated between E14 and E20, the ipsilaterally projecting ganglion cells ceasing their neurogenesis a full day before the contralaterally projecting ganglion cells. Alpha cells were generated from the very outset of retinal ganglion cell genesis, at E14, but completed their neurogenesis before the other cell types, by E17. Tritium-positive, horseradish peroxidase-negative neurons in the ganglion cell layer were present from E14 through to E22, and are interpreted as displaced amacrine cells. Choline acetyltransferase-positive displaced amacrine cells were generated between E16 and E20. Individual cell types showed a rough centroperipheral neurogenetic gradient, with the dorsal half of the retina slightly preceding the ventral half. These results demonstrate, first, that retinal ganglion cell genesis and displaced amacrine cell genesis overlap substantially in time. They do not occur sequentially, as has been commonly assumed. Second, they demonstrate that the alpha cell population of retinal ganglion cells and the choline acetyltransferase-immunoreactive population of displaced amacrine cells are each generated over a limited time during the periods of overall ganglion cell and displaced amacrine cell genesis, respectively. Third, they show that the very earliest ganglion cells to be generated in the temporal retina have exclusively uncrossed optic axons, while the later cells to be generated therein have an increasing propensity to navigate a crossed chiasmatic course.