Alcohol effects on lipid bilayer permeability to protons and potassium: relation to the action of general anesthetics

Although general anesthetics are clinically important and widely used, their molecular mechanisms of action remain poorly understood. Volatile anesthetics such as isoflurane (ISO) are thought to alter neuronal function by depressing excitatory and facilitating inhibitory neurotransmission through direct interactions with specific protein targets, including voltage-gated sodium channels (Nav). Many anesthetics alter lipid bilayer properties, suggesting that ion channel function might also be altered indirectly through effects on the lipid bilayer. We compared the effects of ISO and of a series of fluorobenzene (FB) model volatile anesthetics on Nav function and lipid bilayer properties. We examined the effects of these agents on Nav in neuronal cells using whole-cell electrophysiology, and on lipid bilayer properties using a gramicidin-based fluorescence assay, which is a functional assay for detecting changes in lipid bilayer properties sensed by a bilayer-spanning ion channel. At clinically relevant concentrations (defined by the minimum alveolar concentration), both the FBs and ISO produced prepulse-dependent inhibition of Nav and shifted the voltage dependence of inactivation toward more hyperpolarized potentials without affecting lipid bilayer properties, as sensed by gramicidin channels. Only at supra-anesthetic (toxic) concentrations did ISO alter lipid bilayer properties. These results suggest that clinically relevant concentrations of volatile anesthetics alter Nav function through direct interactions with the channel protein with little, if any, contribution from changes in bulk lipid bilayer properties. Our findings further suggest that changes in lipid bilayer properties are not involved in clinical anesthesia.

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Revisiting lipid general anesthetic interactions (II): Halothane location and changes in lipid bilayer microenvironment monitored by fluorescence

Canadian Journal of Chemistry ◽

10.1139/v04-024 ◽

2004 ◽

Vol 82 (7) ◽

pp. 1139-1149 ◽

Cited By ~ 20

Author(s):

Anna Carnini ◽

Heather A Phillips ◽

Leanne G Shamrakov ◽

David T Cramb

Keyword(s):

Phase Transition ◽

Lipid Bilayer ◽

Acyl Chain ◽

Significant Shift ◽

General Anesthetics ◽

General Anesthetic ◽

Scanning Calorimetry ◽

Anesthetic Action ◽

Steady State Fluorescence Spectroscopy ◽

Universal Mechanism

A universal mechanism for the action of general anesthetics (GA) is not yet available. In this study, we investigated the interaction between halothane and 1,2-dipalmitoyl-sn-3-glycero-3-phosphocholine (DPPC) and 1,2-dioleoyl-sn-3-glycero-3-phosphocholine (DOPC) bilayers labeled with Laurdan, Prodan, and NBD-C6-PC as the reporter probes using steady-state fluorescence spectroscopy. We have evidence that halothane is located on the acyl chain side, near the headgroup region of the bilayer. Additionally, we find that halothane may be inhomogeneously distributed within DOPC and DPPC bilayers. We also show data that indicate halothane increases the free volume available to fluorescent probes. Differential scanning calorimetry and UV scanning calorimetry experiments were implemented to further observe the effects of halothane addition to the DPPC lipid bilayer. A significant shift of the phase-transition temperature of the DPPC system was observed. Our findings suggest that general anesthetic lipid bilayer interactions may play a significant role in the overall mechanism of anesthetic action, and these effects should not be ignored when interactions between membrane proteins and anesthetics are considered.Key words: liposomes, anesthesia, fluorescence, phase transition, phospholipid bilayers.

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