In the early phases of the process, this often resulted in reabsorption of the Temozolomide neurite by the cell soma which was followed by the creation of a new initiation site and the outgrowth of a new neurite. In contrast, on the line pattern, neurites almost never retracted and thus outgrowth was steady. We tracked neurite tip trajectories and found that neurite outgrowth on plain substrate typically occurred for a period of 30 min before a retraction event occurred. This neurite extension lifetime was extended to 180 minutes on the line substrate with retraction events typically occurring at neurite branch points. This allowed for the elimination of the branch points and led the cell to adopt two unbranched neuronal processes that align in the direction of the line pattern. We found that neurite tip velocity was only modestly increased on the line versus plain substrate. Soma motility was also affected. On plain substrate, the soma displayed a highly motile behavior consisting of random bursts of migratory behavior. On the line substrate, cells were much less motile. Thus, the line substrate not only allows neurite orientation, but also switches off the dynamic unstable behavior of neurites and the motile behavior of cells observed on plain substrate. The most marked differences in Evofosfamide 918633-87-1 morphological responses of neuronal like cells in response to the plain versus the line pattern are observed at the level of the filopodia which have been proposed to work as sensors to guide neuronal growth cones. Thus, we performed high resolution time-lapse microscopy experiments in which we visualized F-actin dynamics using the Lifeact-GFP probe, which allows for a high contrast on filopodia. On plain substrate, filopodia directly at the growth cone or the neurite shaft extend randomly in multiple directions, perform a typical lateral back and forth motion and then retract. This is accompanied with dynamic neurite protrusion/ retraction cycles in multiple directions as described above. On the line substrate, we found that the two growth cone filopodia populations displayed different dynamic behaviors. Filopodia located at the growth cone tip that aligned on the ridges were stable and contained high amounts of F-actin reflected by elevated Lifeact- GFP signal, compared to the non-aligned filopodia. Nonaligned filopodia situated on the distal part of the growth cone and throughout the neurite shaft displayed a highly unstable behavior and contained less F-actin.