Effect of anaesthetics on calcium-induced luminescence of aequorin

The effect of some general and local anaesthetics on the calcium-induced luminescence of aequorin was studied in vitro using a photomultiplier tube and recording technique. Purified aequorin (0.1 microliter) was injected into a 500 micron diameter porous cellulose acetate capillary tube containing 0.5 M KC1, 20 mM phosphate (pH 7.2) and calcium-EGTA buffers. The trapped aequorin was superfused with buffer solutions which sometimes contained anaesthetic (test) solutions. The results showed that some anaesthetics, e.g. urethane, etomidate and lignocaine, increased whereas others, e.g. methohexitone, thiopentone, decreased the light output (luminescence) of aequorin in constant ionized calcium and EGTA buffers. Similar results were produced by some non-anaesthetic drugs, e.g. glycerol, TEA, caffeine, etc. Concentration-response curves for calcium-dependent and -independent luminescence of aequorin showed that anaesthetics variously affected the aequorin response. Some anaesthetics, e.g. lignocaine, increased the maximum response while others, e.g. etomidate, increased the affinity (i.e. decreased EC50s) of aequorin to calcium ions without altering the slope, which remained at about 2. It was concluded that anaesthetics can either excite or depress aequorin luminescence, the effect being dependent on the type and the concentration used.