Plasma membrane Ca2+-ATPase isoforms in frog crista ampullaris: identification of PMCA1 and PMCA2 specific splice variants.

Ca2+ ions play a pivotal role in inner ear hair cells as they are involved from the mechano-electrical transduction to the transmitter release. Most of the Ca2+ that enters into hair cells via mechano-transduction and voltage-gated channels is extruded by the plasma membrane Ca2+-ATPases (PMCAs) that operate in both apical and basal cellular compartments. Here, we determined the identity and distribution of PMCA isoforms in frog crista ampullaris: we showed that PMCA1, PMCA2 and PMCA3 are expressed, while PMCA4 appears to be negligible. We also identify PMCA1bx, PMCA2av and PMCA2bv as the major splice variants produced from PMCA1 and PMCA2 genes. PMCA2av appears to be the major Ca2+-pump operating at the apical pole of the cell, even if PMCA1b is also expressed in the stereocilia. PMCA1bx is, instead, the principal PMCA of hair cell basolateral compartment, where it is expressed together with PMCA2 (probably PMCA2bv) and PMCA3. Frog crista ampullaris hair cells lack a Na/Ca exchanger, therefore PMCAs are the only mechanism of Ca2+ extrusion. The coexpression of specific isozymes in the different cellular compartments responds to the need of a fine regulation of both basal and dynamic Ca2+ levels at the apical and basal pole of the cell.

Potassium currents in the hair cells of vestibular epithelium: position-dependent expression of two types of A channels.

The complement of voltage-dependent K+ currents was investigated in hair cells of the frog crista ampullaris. The currents were recorded in transversal slices of the peripheral, intermediate and central regions of the crista by applying the patch clamp technique to cells located at different positions in the slices. Voltage-clamp recordings confirmed that cells located in each region have a distinctive complement of K+ channels. Detailed investigation of the currents in each region revealed that the complement of K+ channels in intermediate and central regions showed no variations among cells, whereas peripheral hair cells differed in the expression of two classes of A-type currents. These currents showed different kinetics of inactivation as well as steady-state inactivation properties. We termed these currents fast I(A) and slow I(A) based on their inactivation speed. The magnitude of both currents exhibited a significant gradient along the transversal axis of the peripheral regions. Fast I(A) magnitude was maximal in cells located in the external zone of the crista slice and decreased gradually to become very small in the median zone (centre) of the section, while the gradient of slow I(A) magnitude was reversed. A-type currents appear to act as a transient buffer that opposes hair cell depolarization induced by positive current injections. However, fast I(A) is partially active at the cell resting potential, while slow I(A) can be recruited only following large hyperpolarizations. Thus, two types of A currents are differentially distributed in vestibular hair cells and have different roles in shaping receptor potential.