M Channels Containing KCNQ2 Subunits Modulate Norepinephrine, Aspartate, and GABA Release from Hippocampal Nerve Terminals

• Published in: The Journal of neuroscience 2004-01, Vol.24 (3), p.592-597
• Main Authors: Martire, Maria, Castaldo, Pasqualina, D'Amico, Monia, Preziosi, Paolo, Annunziato, Lucio, Taglialatela, Maurizio
• Format: Article
• Language: English
• Subjects: Aminopyridines - pharmacology; Animals; Anthracenes - pharmacology; Anticonvulsants - pharmacology; Aspartic Acid - chemistry; Aspartic Acid - metabolism; Brief Communications; Carbamates - pharmacology; CHO Cells; Cricetinae; gamma-Aminobutyric Acid - chemistry; gamma-Aminobutyric Acid - metabolism; Hippocampus - chemistry; Hippocampus - metabolism; KCNQ2 Potassium Channel; KCNQ3 Potassium Channel; Male; Norepinephrine - chemistry; Norepinephrine - metabolism; Patch-Clamp Techniques; Phenylenediamines - pharmacology; Potassium - metabolism; Potassium Channel Blockers - pharmacology; Potassium Channels - chemistry; Potassium Channels - drug effects; Potassium Channels - metabolism; Potassium Channels, Voltage-Gated; Presynaptic Terminals - chemistry; Presynaptic Terminals - metabolism; Protein Subunits - chemistry; Protein Subunits - metabolism; Rats; Synaptosomes - chemistry; Synaptosomes - drug effects; Synaptosomes - metabolism
• ISSN: 0270-6474; 1529-2401
• DOI: 10.1523/JNEUROSCI.3143-03.2004

KCNQ subunits encode for the M current (I(KM)), a neuron-specific voltage-dependent K+ current with a well established role in the control of neuronal excitability. In this study, by means of a combined biochemical, pharmacological, and electrophysiological approach, the role of presynaptic I(KM) in the release of previously taken up tritiated norepineprine (NE), GABA, and d-aspartate (d-ASP) from hippocampal nerve terminals (synaptosomes) has been evaluated. Retigabine (RT) (0.01-30 microm), a specific activator of I(KM), inhibited [3H]NE, [3H]d-ASP, and [3H]GABA release evoked by 9 mm extracellular K+ ([K+]e). RT-induced inhibition of [3H]NE release was prevented by synaptosomal entrapment of polyclonal antibodies directed against KCNQ2 subunits, an effect that was abolished by antibody preabsorption with the KCNQ2 immunizing peptide; antibodies against KCNQ3 subunits were ineffective. Flupirtine (FP), a structural analog of RT, also inhibited 9 mm [K+]e-induced [3H]NE release, although its maximal inhibition was lower than that of RT. Electrophysiological studies in KCNQ2-transfected Chinese hamster ovary cells revealed that RT and FP (10 microm) caused a -19 and -9 mV hyperpolarizing shift, respectively, in the voltage dependence of activation of KCNQ2 K+ channels. In the same cells, the cognition enhancer 10,10-bis(4-pyridinylmethyl)-9(10H)-anthracenone (XE-991) (10 microm) blocked KCNQ2 channels and prevented their activation by RT (1-10 microm). Finally, both XE-991 (10-100 microm) and tetraethylammonium ions (100 microm) abolished the inhibitory effect of RT (1 microm) on [3H]NE release. These findings provide novel evidence for a major regulatory role of KCNQ2 K+ channel subunits in neurotransmitter release from rat hippocampal nerve endings.