Growth cone morphology varies with position in the developing mouse visual pathway from retina to first targets

Abstract

We have labeled the growth cones of retinal ganglion cell axons with HRP in intact mouse embryos. This has allowed us to visualize growth cone morphology during outgrowth along an entire CNS pathway from origin to target; to ask whether growth cone forms, and thus behaviors, differ at various points along the pathway; and to study the relationships of growth cones with the cellular environment. During the major period of axon outgrowth between embryonic day (E) 12 and 15, growth cones in the optic nerve are highly elongated (up to 40 microns) and have lamellopodial expansions, but the majority lack the microspikes or filopodia characteristic of many growth cones. Within the optic chiasm (E13-15), most growth cones shorten and spread, and project several short filopodia. In the optic tract, growth cones become more slender and again lack filopodia, resembling sleeker versions of optic nerve growth cones. Near the first target region (lateral geniculate nucleus), growth cones with filopodia arise from individual axon lengths and turn medially toward the target. Within target regions, the branches of immature axon arbors are tipped by minute swellings rather than by the enlarged growth cones prevalent during outgrowth toward targets. Electron-microscopic analysis of identified labeled growth cones in the optic nerve reveal intimate interactions between growth cones and glia or other growth cones in the form of invaginating contacts. In the optic nerve, growth cones contact immature glial (neuroepithelial) cells somewhere along their length, and also envelop bundles of neurites. In the chiasm, single growth cones simultaneously relate to many different profiles. These results demonstrate that in this single pathway from origin to targets, growth cone morphology varies systematically with position along the visual pathway. During outgrowth, simple growth cones are prominent when axons follow well-defined common pathways, and more elaborate filopodial forms appear when growth cones diverge, as they turn or come to decision regions. Together with observations in vitro and in nonmammalian nervous systems in situ, these data serve as reference points for testing to what extent growth cone form reflects intrinsic factors and interactions with the environment.