The localization of alpha-atrial natriuretic polypeptide (alpha-ANP) in the rat cochlea was studied by immunohistochemical technique, using a polyclonal antibody against synthetic rat alpha-atrial natriuretic polypeptide (alpha-rANP). The spiral ligament of the lateral cochlear wall exhibited pronounced immunoreactivity to atrial natriuretic polypeptide (ANP), whereas the stria vascularis displayed almost no immunoreaction. ANP immunoreactivity (IR) was also intense in the spiral limbus region. IR was observed in the fibroblast-like cells and in the extracellular matrix, in particular along its fibrous bundles, of both the spiral ligament and the spiral limbus. These results indicate that ANP may participate in the regulation of the water electrolyte balance at these sites, which may imply a role for ANP in the regulation of cochlear fluids.