Ventral tegmental area (VTA) GABA neurons are critical substrates modulating the mesocorticolimbic dopamine system implicated in natural and drug reward. The aim of this study was to evaluate the effects of ethanol on glutamatergic and GABAergic modulation of VTA GABA neuron electrical synaptic transmission. We evaluated the effects of systemic ethanol (0.05-2.0 g/kg i.p.), the N-methyl-D-aspartate (NMDA) receptor antagonist dizocilpine (MK-801; 0.05-0.2 mg/kg i.v.), the connexin-36 gap junction blocker quinidine (5-20 mg/kg i.v.), the fast-acting barbiturate methohexital (Brevital; 5-10 mg/kg i.v.), and the benzodiazepine chlordiazepoxide (Librium; 5-10 mg/kg i.v.), as well as in situ VTA administration of NMDA and the GABA A receptor agonist muscimol, on VTA GABA neuron spontaneous activity and internal capsule stimulus-induced poststimulus spike discharges (ICPSDs). Systemic ethanol, quinidine, and dizocilpine reduced, whereas local NMDA enhanced, and the systemic and local GABA A receptor modulators did not significantly alter VTA GABA neuron ICPSDs. Ethanol potentiated dizocilpine inhibition of VTA GABA neuron ICPSDs, but not quinidine inhibition. In situ microelectrophoretic application of dopamine markedly enhanced VTA GABA neuron firing rate (131%), spike duration (124%), and spike coupling, which were blocked by systemic quinidine. These findings indicate that VTA GABA neurons are coupled electrically via gap junctions and that the inhibitory effect of ethanol on electrical transmission is primarily via inhibition of NMDA receptor-mediated excitation, not via enhancement of GABA receptor-mediated inhibition. Thus, the rewarding properties of ethanol may result from inhibitory effects on excitatory glutamatergic neurotransmission between electrically coupled networks of midbrain GABA neurons.The two major types of membrane-bound proteins that are directly affected by physiologically relevant levels of ethanol (i.e., concentrations up to 100 mM or 460 mg/dl, at which point ethanol can be lethal in humans) are ligand-gated ion channels and voltage-dependent calcium channels (Harris, 1999). Ligand-gated ion channels, including ␥-GABA, N-methyl-D-aspartate (NMDA), glycine, nicotinic cholinergic, and 5-hydroxytryptamine type 3 receptors play a major role in synaptic transmission and have been shown to be directly modulated by ethanol. In particular, the intoxicating and rewarding properties of ethanol seem to result from either attenuation of NMDA receptor-mediated and/or enhancement of GABA receptor-mediated neurotransmission (for reviews, see Chester and Cunningham, 2002;Davies, 2003).The ventral tegmental area (VTA) is the neuronal origin of the mesocorticolimbic dopamine projection and has been implicated in locomotor activity, cognition, and in the reinforcing/rewarding properties of drugs of abuse (Wise, 1996), including ethanol. With drugs such as cocaine and amphetamine, the link to dopamine systems is readily apparent; however, with other drugs such as opiates and ethanol, the link to dopamin...

Communication between neurons in the mammalian brain is primarily through chemical synapses; however, evidence is accumulating in support of electrical synaptic transmission between some neuronal types in the mature nervous system. The authors have recently demonstrated that the gap junction (GJ) blocker quinidine suppresses stimulus-induced and dopamine-evoked coupling of gamma amino butyric acid (GABA) neurons in the ventral tegmental area (VTA) of mature rats (Stobbs et al., 2004). The aim of this study was to evaluate the role of connexin-36 (Cx36) GJs in mediating electrical coupling between VTA GABA neurons in P50-80 rats in vivo and P25-50 rats in vitro. Single stimulation of the internal capsule (IC) evoked VTA GABA neuron spike couplets in mature rats when activated antidromically, and multiple poststimulus spike discharges (PSDs) when activated with brief high-frequency stimulation of the IC (ICPSDs). The Cx36 GJ blocker mefloquine (30 mg/kg) suppressed VTA GABA neuron ICPSDs in mature freely behaving rats. VTA GABA neurons recorded via whole-cell patch clamp in the midbrain slice preparation of P25-50 rats showed robust expression of Cx36 transcripts when tested with single-cell quantitative reverse transcription polymerase chain reaction. In P50-80 rats, Cx36 protein immunoreactivity was evident in the VTA and surrounding structures. Dye-coupling between VTA neurons was observed following Neurobiotin labeling of VTA GABA neurons, as well as with the fluorochrome Alexa Fluor 488 using real-time video fluorescent microscopy. Thus, mature VTA GABA neurons appear to be connected by electrical synapses via Cx36 GJs, whose coupling is enhanced by corticotegmental input and by dopamine.

Normally, cell membranes resist hydrolysis by secretory phospholipase A 2 . However, upon elevation of intracellular calcium, the cells become susceptible. Previous investigations demonstrated a possible relationship between changes in lipid order caused by increased calcium and susceptibility to phospholipase A 2 . To further explore this relationship, we used temperature as an experimental means of manipulating membrane physical properties. We then compared the response of human erythrocytes to calcium ionophore at various temperatures in the range of 20-50 °C using fluorescence spectroscopy and two-photon fluorescence microscopy. The steady state fluorescence emission of the environmentsensitive probe, laurdan, revealed that erythrocyte membrane order decreases systematically with temperature throughout this range, especially between 28 and 45 °C. Furthermore, the ability of calcium ionophore to induce increased membrane order and susceptibility to phospholipase A 2 depended similarly on temperature. Both responses to calcium influx were enhanced as membrane fluidity increased. Analysis of the spatial distribution of laurdan fluorescence at several temperatures indicated that the ordering effect of intracellular calcium on fluid membranes generates an increase in the number of fluid-solid boundaries. Hydrolysis of the membrane appeared to progress outward from these boundaries. We conclude that phospholipase A 2 prefers to hydrolyze lipids in fluid regions of human erythrocyte membranes, but primarily when those regions coexist with domains of ordered lipids.

There is an ongoing controversy regarding the role of gender in modulating components of the human visual-evoked potential (VEP) and event-related potentials (ERPs). Our aim was to further characterize the role of gender on VEPs, ERPs and response performance in an object recognition task. We recorded VEPs and reaction time (RT) in a paradigm wherein subjects responded to a randomly presented "Relevant" stimulus, and did not respond when presented with "Irrelevant" or "Standard" visual stimuli. There was no effect of gender on early components of the VEP or RT to Relevant stimuli. Relevant and Irrelevant stimuli evoked distinct VEP components including the P300, N400 and late-positive (LP) ERPs that were well-discriminated from those of the Standard stimulus. Females were characterized by greater P300 and N400 responses than males for the Relevant stimulus, but exclusively greater N400 responses for the Irrelevant stimulus. There were no significant gender differences for the LP, or for the latency of any ERP component. Gender differences were not attributed to hemispheric asymmetry, as there were no significant differences in P300 and N400 VEP amplitudes between lateral occipital or parietal electrode positions. These results indicate that the N400 can be elicited in a task requiring the processing of irrelevant, but not unexpected, stimuli and that females process visual information differently than males, perhaps by increased allocation of attentional resources to distracting stimuli.