Adenosine 5′-triphosphate (ATP) concentrations in the endolymph and perilymph of the guinea-pig cochlea

Abstract

The concentration of adenosine 5′-triphosphate (ATP) in endolymph (EL), perilymph (PL) and cerebrospinal fluid (CSF), collected from anesthetized guinea pigs was determined using the luciferase-luciferin reaction. The cochlea was exposed by a ventrolateral approach and the bone overlying scala media of the third turn (EL) or scala vestibuli of the first turn (PL) was shaved to a thin layer and a small fenestrum made. For EL sampling, a double-barrelled pipette was inserted through the spiral ligament-stria vascularis complex. One barrel was filled with 150 mM KCl to record the endocochlear potential (EP) and upon the appearance of the positive EP, 0.12–1.22 μl of fluid was aspirated into the other barrel by gentle negative pressure. For PL sampling, a single-barrelled pipette was advanced into scala vestibuli and 0.3–1.6 μl of fluid was collected by capillarity. CSF (0.36–1.75 μl) was obtained from the cisterna magna. The cochleae were removed and processed for light microscopy to determine the extent of tissue damage from the sampling procedure. ATP concentrations (mean ± SEM, nM) for EL, PL and CSF were 12.95±2.4 ( n = 10), 10.5 ± 3.9 ( n = 11) and 16.1 ± 5.4 ( n =11) respectively. Differences in ATP concentrations among fluids were not statistically significant. To test the effect of hypoxia on ATP levels, a group of guinea pigs was subjected to a90 s period of respiratory anoxia prior to sampling of EL, PL or CSF. ATP concentrations were 14.4 ± 3.5 ( n = 11), 20.7 ± 4.1 ( n = 10) and 13.5 ± 4.6 ( n = 4) for EL, PL and CSF, respectively; only PL ATP concentrations were statistically different ( P = 0.018, Wilcoxon rank sum test) to basal conditions. This is the first study which demonstrates the presence of free ATP in cochlear fluids. The results indicate that ATP is present in cochlear fluids at concentrations close to those known to cause hair cell depolarization in vitro.