Adhesion-clutch drives three-dimensional axon outgrowth

Axon outgrowth requires forces generated by the growth cone. A key model explaining this force generation is the adhesion-clutch mechanism, in which adhesion and clutch molecules convert backward movement of actin filaments into the force that drives axon outgrowth. However, this mechanism has not been validated in three-dimensional (3D) environments. Additionally, a recent study reported that inhibiting actin dynamics or the cell adhesion molecule integrin did not affect axon outgrowth in a 3D collagen gel, challenging the adhesion-clutch paradigm. Here, we show that the adhesion molecule N-cadherin and the clutch molecule shootin1a form a non-integrin adhesion-clutch in a 3D environment containing an appropriate adhesive substrate, N-cadherin. We detected forces produced by growth cones when N-cadherin was present. Furthermore, inhibition of N-cadherin, shootin1a or actin dynamics suppressed 3D axon outgrowth. Our findings demonstrate that the adhesion-clutch is critical machinery for 3D neural network formation under the regulation of specific adhesions.