Abstract
The precise wiring of the nervous system relies on neurons extending their processes at the right time and place to find their appropriate synaptic partner. The mechanisms that determine when and where neurons extend their neurites during synaptogenesis remains a central question in the field. In the present study, we developed a cell culture system coupled with live imaging to investigate the wiring mechanisms in the developing nervous system. We focused on horizontal cells which are interneurons in the mammalian outer retina known to synapse selectively to distinct photoreceptors. Our data shows cultured horizontal cells extend neurites in a similar manner as in vivo with horizontal cells isolated from young mice extending more complex processes compared to those from adult retinas. In addition, horizontal cells cultured alone do not extend neurites and require other retinal cells for neurite extension suggesting that there must be extrinsic cues that promote neurite outgrowth. Moreover, these extrinsic cues do not appear to be solely secreted factors as supernatant from wild-type retinas is not sufficient to promote neurite outgrowth. In summary, we established a new system that can be used to decipher the mechanisms involved in neuronal wiring of the developing central nervous system.