Enteric neurons, controlling various gut functions, are prone to oxidative insults, which might damage mitochondria (eg. intestinal inflammation). To resume local energy supply, mitochondria need to be transported. We used MitoTracker dyes and confocal microscopy to investigate basic characteristics of mitochondrial transport in guinea-pig myenteric neurites. During a 10 second observation of 1 millimeter nerve fiber, on average 3 mitochondria were transported at an average speed of 0.41 ± 0.02 µm.s^-1. Movement patterns were clearly erratic and velocities were independent of mitochondrial size. The velocity oscillated periodically (~6s) but was not consistently affected by structures such as en route boutons, bifurcations or stationary mitochondria. Also, mitochondria transported in opposite directions did not necessarily affect each others mobility. Transport was blocked by microtubule disruption (colchicine, 100µM) and destabilization (cytochalasin-D, 1µM) or stabilization (phalloidin, 10µM) of actin filaments respectively decreased (0.22±0.02 µm.s^-1, p<0.05) or increased (0.53±0.02 µm.s^-1, p<0.05) transport speed. Transport was inhibited by TTX (1µM) and removal of extracellular Ca²⁺ (+ 2mM EGTA) had no effect. However, depletion of intracellular stores (thapsigargin) reduced (to 33%) and slowed the transport significantly (0.18±0.02 µm.s^-1, p<0.05), suggesting an important role for stored Ca²⁺ in mitochondrial transport. Transport was also reduced (to 21%) by the mitochondrial uncoupler FCCP (1µM) in a time-dependent fashion and slowed by oligomycin (10µM). We conclude that mitochondrial transport is remarkably independent of structural nerve fiber properties. We also show that mitochondrial transport is TTX-sensitive; speeds up by stabilizing actin and that functional Ca²⁺ stores are required for efficient transport.