When microtubules in teleost melanophores are disrupted with antimitotic agents, colchicine, high hydrostatic pressure, low temperature, and vinblastine, the alignment and movement of the pigment granules in these cells disappear; during recovery, the return of alignment and movement corresponds in both time and space with the repolymerization of microtubules. Furthermore, analysis of nearest neighbor distances in untreated melanophores reveals that pigment granules are closely associated with microtubules. Other structures such as microfilaments, the endoplasmic reticulum, and the cytoplasmic matrix do not appear to be involved. Thus we conclude that microtubules determine the alignment and are essential for the selective movements of the pigment granules in these cells. Investigations of the mechanism of movement show that microtubules are required for both centrifugal and centripetal migrations and that they do not change in number or location during redistribution of pigment. Our results further indicate that microtubules in melanophores behave as semistable organelles as determined by investigation with colchicine and hydrostatic pressure. These observations and others rule out a push-pull mechanism based on the polymerization and depolymerization of microtubules or one which distinguishes two operationally different sets of microtubules. We propose instead that particles move by sliding along a fixed array of microtubules.