Cross-species collapse activity of polarized radial glia on retinal ganglion cell axons.

Abstract

Inhibition of incorrect axonal outgrowth has been shown to be a crucial guidance mechanism during the development of the nervous system. Within the visual system of chick and rat, extension of retinal ganglion cell axons is essentially restricted to distinct layers of the retina and distinct brain regions such as the tectum opticum. In addition, populations of ganglion cells from defined retina locations project topographically to defined tectal areas, their growth possibly being inhibited by radial glia in incorrect tectal regions. In the current study, we aimed to analyse potential inhibitory activity of retinal glia during outgrowth of ganglion cell axons of embryonic chick and rat. The response of ganglion cell axons originating from different retina locations when exposed to purified retinal radial glia cell membranes were monitored in collapse assays by time lapse video recording. The interaction of axons growing on purified glial somata or glial endfeet was analysed in outgrowth assays. Our results indicate that (1) nasal and temporal chick growth cones are equally induced to collapse by cell membranes from retinal radial glia: 75% nasal and 72% temporal. (2) The collapse inducing component of radial glia can be inactivated by defined heat treatment, reducing collapsing activity to 6% nasal and 5% temporal. (3) Rat growth cones respond in a similar way to chick radial glia. (4) Rat axons grow perfectly on endfeet but not on somata of radial glia of the chick. In summary, the data suggest that radial glia are functionally polarized with permissive endfeet and inhibitory somata based on heat-labile proteins. Glia polarization is likely to inhibit aberrant growth of ganglion cell axons into outer retina layers. However, retinal radial glia are unlikely to participate in preordering axons within the retina and therefore do not affect the topographic projection. Finally, the inhibitory function of radial glia is conserved between birds and mammals and represents possibly a fundamental mechanism for structuring the central nervous system.