The permissive cue laminin is essential for growth cone turning in vivo

The proper guidance of migrating growth cones relies on the balance of multiple guidance cues in the embryonic environment. In addition to guidance cues, growth cones are in contact with other substrates that may contribute to the pathfinding of neurons. For example, in the developing insect peripheral nervous system, pioneer neurons migrate on and between layers of the basal lamina. Previous studies have demonstrated that one basal lamina molecule, laminin, promotes outgrowth of many classes of neurons in vitro. In this study, the simple grasshopper nervous system was used to investigate the role of laminin in neuronal pathfinding. Laminin expression precedes axonogenesis of the Tibial (Ti1) pioneer neurons in the developing limb bud, and expression continues during outgrowth and guidance of the pioneer neurons. The role of a nidogen-binding motif on laminin was investigated using subunit-specific antibodies and peptides as blocking reagents in vivo. Antibodies and peptides that block the nidogen-binding site on laminin resulted in stalled Ti1 axon migration, predominantly at the precise location where they normally turn ventrally. After prolonged culturing, Ti1 axons remained stalled at the same location. Therefore, although Ti1 axons were capable of outgrowth in the presence of blocking reagents, they were not able to navigate an essential turn. This study indicates that the interaction of the Ti1 growth cone with the nidogen-binding site on laminin is vital for neuronal pathfinding in vivo and suggests that permissive cues may be essential for growth cone steering.