Inhibitory transmission in locus coeruleus neurons expressing GABAA receptor epsilon subunit has a number of unique properties.

Fast inhibitory synaptic transmission in the brain relies on ionotropic GABA(A) receptors (GABA(A)R). Eighteen genes code for GABA(A)R subunits, but little is known about the epsilon subunit. Our aim was to identify the synaptic transmission properties displayed by native receptors incorporating epsilon. Immunogold localization detected epsilon at synaptic sites on locus coeruleus (LC) neurons. In situ hybridization revealed prominent signals from epsilon, and mRNAs, some low beta1 and beta3 signals, and no gamma signal. Using in vivo extracellular and in vitro patch-clamp recordings in LC, we established that neuron firing rates, GABA-activated currents, and mIPSC charge were insensitive to the benzodiazepine flunitrazepam (FLU), in agreement with the characteristics of recombinant receptors including an epsilon subunit. Surprisingly, LC provided binding sites for benzodiazepines, and GABA-induced currents were potentiated by diazepam (DZP) in the micromolar range. A number of GABA(A)R ligands significantly potentiated GABA-induced currents, and zinc ions were only active at concentrations above 1 muM, further indicating that receptors were not composed of only alpha and beta subunits, but included an epsilon subunit. In contrast to recombinant receptors including an epsilon subunit, GABA(A)R in LC showed no agonist-independent opening. Finally, we determined that mIPSCs, as well as ensemble currents induced by ultra-fast GABA application, exhibited surprisingly slow rise times. Our work thus defines the signature of native GABA(A)R with a subunit composition including epsilon: differential sensitivity to FLU and DZP and slow rise time of currents. We further propose that alpha(3,) beta(1/3,) and epsilon subunits compose GABA(A)R in LC.

alpha(2)-Adrenoceptors mediate the acute inhibitory effect of fluoxetine on locus coeruleus noradrenergic neurons.

So far, the mechanisms underlying the action of selective serotonin reuptake inhibitors, such as fluoxetine, are not completely understood. Thus, to clarify if fluoxetine has any effect on noradrenergic transmission, we measured the spontaneous firing rate of noradrenergic neurons in the locus coeruleus both in vivo and in vitro using single-unit extracellular recordings. In anesthetized rats, fluoxetine (2.5-20 mg/kg, i.v.) reduced the firing rate in a dose-dependent manner, reaching a maximal inhibition of 55 +/- 5% with respect to the basal value. This effect was not only completely reversed by the alpha(2)-adrenoceptor antagonist, RX 821002 (0.2 mg/kg, i.v.), but also prevented by previous application of both idazoxan (0.05 and 0.1 mg/kg, i.v.) and RX 821002 (6.25 microg/kg, i.v). Furthermore, when noradrenaline was depleted from axon terminals by means of the injection of alpha-methyl-DL-tyrosine (250 mg/kg, i.p.) 24 h prior to the experiment, fluoxetine failed to inhibit locus coeruleus activity. In rat brain slices, perfusion with fluoxetine (100 microM for 5 min) did not modify the firing rate of locus coeruleus neurons (n = 7). We conclude that fluoxetine inhibits locus coeruleus neurons in vivo through a mechanism involving noradrenaline interacting with alpha(2)-adrenoceptors. However, the lack of effect on brain slices would seem to indicate that afferents to the nucleus may be involved in the observed inhibitory effect.