Crossed and uncrossed retinal axons respond differently to cells of the optic chiasm midline in vitro

In mouse, retinal axon divergence takes place within a cellular specialization localized at the midline of the optic chiasm. To test whether the cells in this locus present cues for differential retinal axon growth, retinal explants were cocultured with cells dissociated from the chiasmatic midline, both taken from day 14-15 embryos, during the principal period of retinal axon divergence. Compared with crossed axons from other retinal regions, axons from ventrotemporal retina, the sole source of uncrossed axons, were shorter, more fasciculated, and fewer in number when growing on chiasm cells. Furthermore, uncrossed axons avoided clusters of chiasm neurons and glia having the composition and arrangement of the midline specialization, but crossed axons readily grew over them. In contrast to the clusters of chiasm cells, however, individual neurons and glia did not elicit differential retinal axon growth. These data demonstrate that cues for divergence derive from cells resident to the chiasm and suggest that cellular interactions among resident midline cells are required to produce these cues.