The tumor-promoting drug 4 beta-phorbol 12-myristate 13-acetate (TPA) causes the loss of myofibrils in primary cultures of chick embryonic myotubes [9]. When myofibrils in chick myotubes are dispersed by TPA treatment (5 X 10(-8) M), there remains a class of non-myofibrillar actin filaments that are sensitive to subsequent breakdown by cytochalasin D. Microfilament bundles in fibroblasts in the same cultures seem unaffected by this TPA treatment, but are also broken down by cytochalasin D (0.2 micrograms/ml); this dose has little effect on myofibrils. We have previously shown that treatment of chick myotubes with cytochalasins would destabilize clusters of acetylcholine receptors (AChRs) [6]. In order to further examine the relationship between actin filaments and cell surface AChRs, we have used the receptor-specific ligand alpha bungarotoxin (a-BGT) to study the fate of AChR clusters in drug-treated and control myotubes. Cells treated with TPA showed no loss in the number of receptor clusters present on their surface. However, if these cells were also treated with cytochalasin D, cluster number was reduced to approximately the same value as seen for cytochalasin treatment alone (50% of the control value). These results suggest that the cytoskeletal link to these cell surface receptors is not mediated by attachment to the alpha actin-containing myofibrils, but rather clustered AChRs are stabilized by a class of non-myofibrillar actin filaments.