T. Munsch et Hc. Pape, Upregulation of the hyperpolarization-activated cation current in rat thalamic relay neurones by acetazolamide, J PHYSL LON, 519(2), 1999, pp. 505-514
1. The effect of inhibition of brain carbonic anhydrase (CB) on the hyperpo
larization-activated cation current (I-h) of thalamocortical (TC) neurones
of the rat ventrobasal thalamic complex (VB) was investigated in an in vitr
o slice preparation using the whole-cell patch-clamp technique and fluoresc
ence ratio imaging of the pH indicator 2',7'-bis(carboxyethyl) -5(and -6)-c
arboxyfluorescein (BCECF).
2. Recording of I-h before and after addition of 0.4-0.8 mM acetazolamide t
o the bathing fluid revealed a significant shift in the voltage dependence
of activation (V-1/2) of 5-7 mV to more positive potentials.
3. Simultaneous recording of I-h and BCECF fluorescence ratio (F-420/F-495)
revealed an increase in I-h amplitude accompanied by an intracellular alka
linization upon application of acetazolamide. The CA inhibitor ethoxyzolami
de (EZA, 50 mu M) also led to an intracellular alkalinization and a subsequ
ent 4-5 mV positive shift of V-1/2 of I-h.
4. Acetazolamide and EZA both profoundly slowed the rapid fall of pH, upon
switching from Hepes- to CO2/HCO3--buffered superfusate, indicating intrace
llular CA isoforms in TC neurones.
5. In slices bathed in Hepes-buffered saline, addition of acetazolamide had
no effect on the amplitude and time course of activation of I-h, indicatin
g that the action of acetazolamide on I-h was dependent on the presence of
HCO3-.
6. Under current-clamp conditions, the neuronal response to hyperpolarizing
current pulses in the presence of acetazolamide was decreased as compared
to control. This resulted in a strongly reduced ability of TC neurones to p
roduce rebound Ca2+-mediated spikes.
7. The present results implied that in TC neurones acetazolamide led to an
intracellular alkalinization which causes, due to its pH sensitivity, an in
crease in I-h.