The authors investigated the effects of chronic kanamycin (KM) administration on the basement membrane (BM) anionic sites in the ampulla by studying the binding of cationic polyethyleneimine (PEI). KM sulfate was administered intramuscularly 'to guinea pigs with normal Preyer's reflexes daily for 10 or 17 days. The PEI distribution was unchanged on the subepithelial BM in the dark cell region and on the capillary BM in the crista ampullaris. However, PEI binding decreased significantly on the subepithelial BM in the sensory cell and transitional cell regions of those guinea pigs administered KM for 17 days. In the sensory cell region, the PEI distribution did not recover until 6 weeks after KM treatment. Findings suggest that chronic administration of KM severely alters the number of subepithelial BM anionic sites in the sensory cell region.