Differentiation of neuronal growth cones: specialization of filopodial tips for adhesive interactions.
Adhesive contacts made by filopodia of developing neurons are important in neurite growth and in the formation of synaptic junctions. In the present work, filopodial interactions of cultured chicken retina neurons were studied by using video-enhanced contrast, differential interference contrast (VEC-DIC) microscopy and the high-voltage electron microscope (HVEM). Use of the HVEM to examine whole mounts of fixed cells showed that filopodia in older cultures developed an appearance that might be expected of nascent synapses, becoming enlarged at their endings and accumulating organelles resembling synaptic vesicles. VEC-DIC microscopy, used to observe the motility and adhesive properties of filopodia in living cells, showed there was a particularly high affinity between filopodia tips. Contacting filopodia typically repositioned themselves so they could attach at a tip-to-tip position, occasionally bending as much as 90 degrees to achieve this preferred orientation. Interacting filopodia frequently remained together as they pushed or pulled on each other, moved laterally together, or stretched tightly and underwent intense vibratory movements. Such linked motility occurred even when apparent gaps existed between the filopodia. Examination of these gaps with the HVEM revealed filamentous structures linking the apposed membranes. The filamentous links were 10-13 nm in diameter and 30-100 nm long. Although it has not yet been established that the filaments reflect the native configuration of the interconnecting materials, the structures seem likely to be associated with the strongly adhesive behavior of the filopodial tips. The possible significance of these structural and functional properties of filopodia tips to axon growth and synapse formation is discussed.
D000818:Animals; D002448:Cell Adhesion; D002454:Cell Differentiation; D002478:Cells, Cultured; D002642:Chick Embryo; D008853:Microscopy / Q000379:methods; D008854:Microscopy, Electron / Q000379:methods; D009040:Motion Pictures; D009474:Neurons / Q000648:ultrastructure*; D012160:Retina / Q000166:cytology; D013569:Synapses / Q000166:cytology*