INTRACELLULAR AND EXTRACELLULAR CALCIUM MODULATES STEREOCILIA STIFFNESS ON CHICK COCHLEAR HAIR-CELLS

Segments of the chick basilar papilla were isolated and maintained in culture medium. The sensory hair bundle of individual hair cells was observed with light microscopy and stimulated with a water microjet at 600 Hz. Hair bundle motion was slowed by illuminating the microscope with stroboscopic light, and water jet intensity was systematically varied in decibel (dB) steps until a visual detection level (VDL) threshold of hair bundle motion was achieved. The VDL threshold of many hair cells was measured in each isolated papilla. However, only one of eight extracellular calcium concentrations (0.0, 0.0001, 0.001, 0.01, 0.1, 1.25, 6.0, and 12.0 mM) was used with each papilla. In a second series, a calcium ionophore (ionomycin) was added to the culture medium, and VDL thresholds were again measured at seven of these extracellular calcium concentrations. With extracellular calcium alone, the stimulus level needed to achieve threshold was reduced by 2.73 dB between 0. 1 and 0.01 mM. This change in threshold represented a 1.37-fold decrease in hair bundle stiffness. When ionomycin was added to the culture medium, a progressively greater stimulus intensity was needed to achieve threshold as calcium concentration increased. The 11.7-dB increase in threshold, with the addition of ionomycin, between 0.0001 and 6.0 mM extracellular calcium was equivalent to a 3.85-fold increase in bundle stiffness. These large changes in hair-bundle stiffness, as a function of the extra- or intracellular calcium environment, may play an important role in the micromechanical behavior of the hair cell during sound simulation.