General anesthetic actions on norepinephrine, dopamine, and gamma-aminobutyric acid transporters in stably transfected cells.
The effects of general anesthetics on neurotransmitter uptake by plasma membrane transporters are controversial. We analyzed the effects of representative volatile and IV general anesthetics on recombinant transporters for norepinephrine (human NET), dopamine (rat DAT), or gamma-aminobutyric acid (rat GAT-1) stably expressed in a porcine kidney cell line (LLC-PK(1)). This approach avoids complicating factors associated with neuronal preparations, such as the involvement of multiple transporters and the indirect effects of membrane potential. At clinical concentrations, human NET was inhibited only by halothane (50% inhibitory concentration [IC(50)] = 0.54 mM), rat DAT was sensitive to both halothane and isoflurane (IC(50) = 0.60 and 0.64 mM, respectively), and rat GAT-1 was insensitive to both volatile anesthetics. Human NET was inhibited in a dose-dependent fashion by propofol (IC(50) = 41 micro M), ketamine (IC(50) = 150 micro M), and etomidate (IC(50) > 200 micro M), but not by pentobarbital. Only propofol inhibited NET at a clinically relevant concentration (5 micro M). Rat DAT was inhibited in a dose-dependent fashion by propofol (IC(50) = 120 micro M), etomidate (IC(50) = 100 micro M), and ketamine (IC(50) = 210 micro M), but not by pentobarbital. None of these anesthetics was predicted to inhibit DAT at concentrations that produce anesthesia. Propofol inhibited rat GAT-1, but only at the largest concentration tested. General anesthetics have drug- and subtype-selective actions on neurotransmitter transporters. We conclude that effects on catecholamine, but not gamma-aminobutyric acid, transporters may contribute to secondary synaptic actions of certain anesthetics but are unlikely to be essential to their anesthetic properties.


CONCLUSIONS
Previous studies have implicated neurotransmitter transporters as targets for general anesthetic effects on synaptic transmission. Recombinant transporters for norepinephrine and dopamine were sensitive to certain volatile and IV anesthetics, whereas gamma-aminobutyric acid transporters were insensitive. These anesthetic- and neurotransmitter-specific effects may underlie some of the secondary effects of general anesthetics.
D018681:Anesthetics, General / Q000494:pharmacology*; D000818:Animals; D002352:Carrier Proteins / Q000037:antagonists & inhibitors*; D050483:Dopamine Plasma Membrane Transport Proteins; D004305:Dose-Response Relationship, Drug; D050485:GABA Plasma Membrane Transport Proteins; D006801:Humans; D018374:LLC-PK1 Cells; D008562:Membrane Glycoproteins; D008565:Membrane Proteins / Q000037:antagonists & inhibitors*; D049990:Membrane Transport Modulators; D026901:Membrane Transport Proteins / Q000037:antagonists & inhibitors*; D009419:Nerve Tissue Proteins; D050484:Norepinephrine Plasma Membrane Transport Proteins; D027361:Organic Anion Transporters; D051381:Rats; D013552:Swine; D027981:Symporters / Q000037:antagonists & inhibitors*; D014162:Transfection
D018681:Anesthetics, General; D002352:Carrier Proteins; D050483:Dopamine Plasma Membrane Transport Proteins; D050485:GABA Plasma Membrane Transport Proteins; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D049990:Membrane Transport Modulators; D026901:Membrane Transport Proteins; D009419:Nerve Tissue Proteins; D050484:Norepinephrine Plasma Membrane Transport Proteins; D027361:Organic Anion Transporters; C493413:SLC6A1 protein, human; C493227:SLC6A2 protein, human; C493214:SLC6A3 protein, human; C493415:Slc6a1 protein, rat; C493226:Slc6a2 protein, rat; C493216:Slc6a3 protein, rat; D027981:Symporters