Neuronal growth cones integrate signals from outgrowth-promoting molecules, e.g., laminin (LN) or fibronectin (FN), and outgrowth-inhibiting molecules, e.g., chondroitin sulfate proteoglycans (CSPGs), to navigate through extracellular matrix (ECM). Sensory neurons on LN typically turn to avoid areas rich in inhibitory CSPGs, whereas neuron-like cells of human origin (SH-SY5Y) preferentially stop/stall. These different behaviors may reflect differences in neuron type, response to outgrowth-promoters, or the mechanisms involved in outgrowth vs. inhibition. We used image analysis to determine the effects of different outgrowth promoters on the response of SH-SY5Y cells to inhibitory CSPGs. LN increased neurite initiation and elongation compared to cells plated either on endogenous matrix or FN. On a patterned substratum consisting of alternating stripes of FN and CSPGs, 59.6 ± 9.3% of SH-SY5Y growth cones turned upon CSPG contact, whereas only 31.9 ± 8.2% of growth cones turned at a LN/CSPG border. Growth cones on LN spread more upon contact with CSPG than growth cones on FN, whereas growth cones on LN or FN not contacting CSPGs were morphologically similar. Because it is known that integrins are involved in outgrowth on promoters, we analyzed integrin expression in response to inhibitory CSPGs in a choice assay. CSPGs did not induce increases or redistribution of several integrin subunits in SH-SY5Y cells. Furthermore, an anti-β1 integrin function-blocking antibody did not alter growth cone behavior at a CSPG border. These results indicate that significant mechanistic differences may exist between outgrowth on homogenous outgrowth promoters and growth cone turning at inhibitory molecules. J. Neurosci. Res. 66:630–642, 2001. © 2001 Wiley-Liss, Inc.