Abstract
1. Using whole-cell recording and confocal immunofluorescent microscopy, we have investigated the differential electrogenic activity, subunit expression and subcellular distribution of the Na(+)-K(+)-ATPase in the lemniscal (ventral) and non-lemniscal (dorsal) pathways of the rat medial geniculate body (MGB) in vitro. 2. Bath application of Na(+)-K(+)-ATPase inhibitors strophanthidin or dihydro-ouabain produced a transient, dose-dependent inward current or membrane depolarization which were significantly larger in dorsal MGB neurones than in ventral cells (45.9 +/- 6.45 vs. 24.3 +/- 4.1 pA; P < 0.05). Electrophysiological and morphometric measurements showed that the dorsal MGB neurones had a significantly lower input conductance and a smaller somata than their ventral counterparts. The level of the resting membrane potential also differed by about 6 mV between the two cell populations, with the dorsal cells being more hyperpolarized (-74.2 +/- 0.6 vs. -67.7 +/- 1.3 mV; P < 0.001). 3. Incubation of enzymatically dissociated MGB neurones with fluorescent monoclonal antibodies against alpha 1-alpha 3 isoforms of Na(+)-K(+)-ATPase showed that both dorsal and ventral cells expressed primarily alpha 3 subunits. Confocal laser scanning revealed, however, that the mean pixel density of alpha 3 fluorescent antibodies in the plasma membrane domain, but not in the cytoplasmic compartment, was about 40% higher in dorsal neurones than in the ventral cells (29.7 +/- 4.7 vs. 16.9 +/- 2.3 grey shadow per pixel; P < 0.05). 4. The above results suggest that the electrogenic activity of the Na(+)-K(+)-ATPase is differentially regulated between lemniscal and non-lemniscal auditory thalami through a mechanism that probably involves differential pump densities in the cell membrane.