Immunocytochemical and quantitative studies of Na +,K +-ATPase distribution in the developing chick cochlea

Abstract

Immunocytochemical methods were used to examine cryosections of the embryonic and neonatal chicken cochlea in order to study the histological distribution of the Na +,K 4-ATPase molecule during maturation. In complementary studies the Na +,K +-ATPase capacity of microdissected freeze-dried substructures of the cochlea was determined fluorometrically. The dark cell of the tegmentum vasculosum exhibited intense immunochemical staining of the convoluted basolateral infoldings. The adjacent light cells demonstrated very little staining. The plasma membrane of the hair cell was also stained as were the first order auditory neurons, including the cell soma. The homogene cells and the supporting cells were unstained. The dark cell was only lightly stained with the antibody at stage 45 but became more intense and selective by the seventh postnatal day. The other cells of the cochlear duct exhibited specific immunofluorescent staining of their plasma membranes from stage 45 onwards and the fluorescent intensity did not change. The tegmentum vasculosum exhibited very high activities of the Na +,K +-ATPase enzyme relative to the other structures of the cochlea. Furthermore, a pronounced gradient of enzymatic activity was detected longitudinally. The proximal tip (or high-frequency end) had a sixteen-fold greater capacity for Na +- and K +-dependent ATP hydrolysis relative to the distal tip (or low-frequency end). The appearance of this enzyme in the tegmentum vasculosum during the development of the cochlea paralleled the known rate of improvement in hearing thresholds.