The proper formation of neural circuits depends on the precise pathfinding of axons, which is mediated by various molecules. While the expression of specific guidance molecules by different cells in the nervous system has been well characterized, the mechanisms underlying the secretion and delivery of these molecules to growing axons remain relatively unexplored.In this study, we used ex vivo live imaging techniques to investigate the guidance of commissural axons in the chicken embryo. Our findings reveal that multiple populations of guidepost cells employ long filopodia, known as cytonemes, to interact with developing neurons. Specifically focusing on the floor plate as an intermediate target, we characterized the nature and dynamics of these protrusions during midline crossing. Our analyses demonstrate that these filopodia contain crucial guidance molecules, including adhesion molecules and signaling molecules, suggesting their involvement in conveying guidance signals to axons.Furthermore, our investigation into Sonic Hedgehog (Shh) signaling within floor plate cytonemes unveiled the necessity of Dispatched-1 for the proper secretion of Shh and subsequent guidance of commissural axons in vivo. Additionally, our experiments involving the silencing of Myosin-X, a key motor component implicated in cytoneme dynamics and signaling, resulted in impaired commissural axon guidance in a non-autonomous manner. Altogether our findings show for the first time the implication of cytoneme-mediated signaling in the formation of neural circuits.