Activity-driven sharpening of the regenerating retinotectal projection: effects of blocking or synchronizing activity on the morphology of individual regenerating arbors

Abstract

Both blocking activity with intraocular tetrodotoxin (TTX) and synchronizing activity with a xenon strobe light (1 Hz) prevent retinotopic sharpening of regenerating optic projection in goldfish (Meyer, 1983; Schmidt, 1985; Cook and Rankin, 1986). In this study, we tested, in both normal and regenerating projections, the effects of these two treatments on individual optic arbors. Arbors were stained via anterograde transport of HRP, drawn in camera lucida from tectal whole mounts, and analyzed for spatial extent in the plane of the retinotopic map, order of branching, number of branch endings, depth of termination, and the caliber of the parent axon. In normal tectum, fine, medium, and coarse caliber axons gave rise to small, medium, and large arbors, which averaged 127 microns, 211 microns and 275 microns in horizontal extent, and terminated at characteristic depths. All three classes averaged roughly 21 branch endings. Optic arbors that regenerated with normal patterns of activity returned to a roughly normal appearance by 6-11 weeks postcrush: the same three calibers of axons gave rise to the same three sizes of arbors at the same depths, but they were much less stratified and well on average about 16% larger in horizontal extent. At this time point, arbors regenerated under TTX or strobe were on the average 71 and 119% larger, respectively, than the control-regenerated arbors (larger in all classes), although they had approximately the same number of branch endings and were equally poorly stratified. Synapses formed under strobe were also normal in appearance. Thus the only significant effect of both strobe and TTX treatment was to enlarge the spatial extent of arbor branches. Arbors that were not regenerating were very slightly (but significantly) enlarged by TTX block of activity or strobe illumination. As previous staining showed that regenerating axons initially make widespread branches and later retract many of those branches (Schmidt, Turcotte, Buzzard, and Tieman, 1988; Stuermer, 1988), the present findings support the idea that blocking activity or synchronizing activity prevents retinotopic sharpening by interfering with the elimination of some of the errant branches.