We studied the effect of chronic treatment with kanamycin on the basement membrane (BM) anionic sites in the cochlea and endolymphatic sac using polyethyleneimine (PEI) as a cationic tracer. Albino guinea pigs weighing 250-300 g received kanamycin (400 mg/kg/day, i.m.) for 10 or 17 consecutive days. The number of BM anionic sites as derived from the PEI area was not affected in Reissner's membrane, spiral prominence, basilar membrane or endolymphatic sac, whereas it was significantly decreased in the stria vascularis and spiral limbus, being more marked in the guinea pigs treated for 17 days than in those treated for 10 days. The number of BM anionic sites in these regions did not recover until 6 weeks after kanamycin treatment. These findings suggest that chronically administered kanamycin may selectively and progressively affect the BM anionic sites in the stria vascularis and spiral limbus, resulting in disruption of a barrier function in the cochlea, and that severely impaired BM anionic sites in the cochlea may not recover.