To determine the organization of the three major cytoskeletal elements of cultured human trabecular meshwork cells (actin filaments, microtubules and intermediate filaments), we employed fluorescence microscopy and stereo-transmission electron microscopy of extracted, S-1 labeled and critical-point dried cells. Morphologic changes resulting from treatment with cytochalasin B, colchicine, taxol and nocodozole were also characterized. Compared with the cynomolgus monkey trabecular cell, morphologic differences in overall cell shape and orientation of both actin filaments and microtubules were noted. However, the responses to cytoskeletal active drugs were quite similar. Taxol, nocodozole and colchicine had a marked effect on microtubule organization, while nocodozole and colchicine had a marked effect on vimentin filament organization. None of these drugs produced marked changes in human trabecular cell shape. In contrast, treatment with the anti-actin drug cytochalasin B resulted in both a marked change in cell shape associated with organizational changes in all three cytoskeletal elements. These studies suggest a central role of actin filaments in determining overall cell shape and cytoskeletal organization in the cultured human trabecular cell.