The effects of acrylamide on fast axonal transport have been measured primarily using the indirect methods of isotope or enzyme accumulation. We report the first direct evaluation of the effects of subchronic acrylamide dosing (150, 300, or 500 mg/kg total dose, i.e., 50 mg/kg, 2x/wk, for 1.5, 3, 5 wk, respectively) on the fast axonal transport motility machinery itself using video-enhanced differential interference contrast optics with digital image processing and computer analysis. Four principle observations were made: (1) Rapid anterograde transport was not affected at any dosage level within 1 wk after cessation of dosing. (2) A high cumulative dosage (500 mg/kg total) of acrylamide or bisacrylamide produced approximately 7-18% decrease in the rate of retrograde transport in both myelinated and unmyelinated axons. (3) Lower dosages of acrylamide (150 or 300 mg/kg total) produced an increase in retrograde transport rates in myelinated axons only. (4) During the "recovery" phase for the 500 mg/kg acrylamide animals (i.e., 3 or 5 wk after the last dosage of acrylamide) the rate of anterograde transport in the myelinated axons was decreased at 3 wk but not at 5 wk, and the rate of retrograde transport in the myelinated axons returned to control levels while the retrograde transport in the unmyelinated axons continued at abnormally slow speeds. The application of this new technique to evaluate the neurotoxic effects of acrylamide provides evidence of dynamic changes in the axonal transport motility machinery itself and differential effects on myelinated versus unmyelinated fibers.