Modulation of locus coeruleus neurons by extra- and intracellular adenosine 5'-triphosphate.

The cell membrane of rat locus coeruleus (LC) neurons is sensitive to both extra- and intracellular ATP. Extracellular ATP or its enzymatically stable analogues activate membrane receptors of the P2 type. These receptors inhibit a persistent potassium current and simultaneously activate a nonselective cationic conductance. The resulting depolarization increases the spontaneous firing rate. A decrease in the concentration of intracellular ATP during hypoxia or hypoglycemia opens ATP-sensitive K+ (KATP) channels of LC neurons. The resulting hyperpolarization depresses the discharge of action potentials and conserved energy. The hypoxia-induced hyperpolarization is additionally due to the release of adenosine from neighboring neurons or glial cells. A certain class of compounds, termed potassium channel openers, also decrease the firing, while sulphonylurea antidiabetics known to block KATP channels increase it. Sulphonylurea antidiabetics antagonize the excitability decrease induced both by potassium channel openers and metabolic damage.

Neuropeptide Y potentiates via Y2-receptors the inhibitory effect of noradrenaline in rat locus coeruleus neurones.

Intracellular recordings were carried out in a pontine slice preparation of the rat brain containing the locus coeruleus (LC). Pressure application of noradrenaline with various pulse durations inhibited the spontaneous frequency of action potentials and hyperpolarized the membrane. Neuropeptide Y (NPY), its C-terminal fragment NPY (16-36) and peptide YY (PYY), at a concentration of 0.1 mumol/l all, potentiated the effect of noradrenaline, while [Leu31, Pro34]NPY (0.1 mumol/l) was inactive. These results are compatible with the presence of Y2-type NPY-receptors at the cell somata of LC neurones.

Effects of potassium channel openers and their antagonists on rat locus coeruleus neurones.

1. Intracellular recordings were obtained from a pontine slice preparation of the rat brain containing the locus coeruleus (LC). Two openers of ATP-sensitive potassium (K(ATP)) channels, RO 31-6930 (10 microM) and cromakalim (100 microM) decreased the spontaneous discharge of action potentials without altering their amplitude or duration. Neither compound changed the resting membrane potential. 2. Of two K(ATP) channel blockers, tolbutamide (300 microM) increased the firing rate, while glibenclamide (3 microM) only tended to do so. In addition, both compounds antagonized the effect of RO 31-6930 (10 microM). Neither glibenclamide (3 microM) nor tolbutamide (300 microM) altered the resting membrane potential. 3. Tetrodotoxin (0.5 microM) depressed the firing, but did not influence the inhibitory action of RO 31-6930 (10 microM). The excitatory amino acid antagonist, kynurenic acid (500 microM), did not change the spontaneous discharge of action potentials. 4. Small shifts (2-4 mV) of the membrane potential by hyper- or depolarizing current injections markedly decreased and increased the firing rate, respectively. 5. Noradrenaline (100 microM) hyperpolarized the cells and decreased their input resistance. This effect was not antagonized by glibenclamide (3 microM) or tolbutamide (300 microM). Ba2+ (2 mM), a blocker of both ATP-sensitive and inwardly rectifying potassium channels, abolished the effects of RO 31-6930 (10 microM) and noradrenaline (100 microM). 6. These data suggest that K(ATP) channels are present on the noradrenergic LC neurones, but are not coupled to alpha 2-adrenoceptors.

Galanin receptors inhibit the spontaneous firing of locus coeruleus neurones and interact with mu-opioid receptors.

Electrophysiological experiments were performed in a pontine slice preparation of rat brain containing the locus coeruleus (LC). The extracellular part of this study showed that galanin (0.003-0.1 mumol/l), [Met5]enkephalin (0.01-10 mumol/l) and noradrenaline (0.1-100 mumol/l) concentration dependently inhibited the firing rate. Noradrenaline (1 and 3 mumol/l) had the same effect both before and during the application of galanin (0.001 or 0.01 mumol/l). Similarly, [Met5]enkephalin (0.03 and 0.1 mumol/l) produced identical inhibition, regardless of the presence or absence of 0.01 mumol/l galanin. Whereas rauwolscine (1 mumol/l) potentiated the effect of galanin (0.03 mumol/l), idazoxan (1 mumol/l) was inactive. In contrast, both naloxone (0.1 mumol/l) and beta-funaltrexamine (0.1 mumol/l) facilitated the galanin-induced inhibition. In the intracellular experiments, galanin (0.3 mumol/l) abolished the spontaneous discharge of action potentials, hyperpolarized the cells and decreased their input resistance. In conclusion, galanin may depress the firing rate by increasing a potassium permeability. Moreover, galanin receptors appear to interact with mu-opioid receptors but not with alpha 2-adrenoceptors.

Depression by neuropeptide Y of noradrenergic inhibitory postsynaptic potentials of locus coeruleus neurones.

Intracellular recordings were performed in a pontine slice preparation of the rat brain containing the locus coeruleus (LC). The spontaneous firing of action potentials was prevented by passing continuous hyperpolarizing current via the recording electrode. Focal electrical stimulation evoked a synaptic depolarization (PSP) followed by a hyperpolarization (IPSP). Neuropeptide Y (NPY; 0.1 mumol/l) inhibited the IPSP only. Pressure ejection of noradrenaline produced hyperpolarization which was potentiated in the presence of NPY (0.1 mumol/l). Hence, NPY appears to inhibit the release of noradrenaline from dendrites or recurrent axon collaterals of LC neurones.

Depolarization of rat locus coeruleus neurons by adenosine 5'-triphosphate.

Intracellular recordings were performed in a pontine slice preparation of the rat brain containing the locus coeruleus. The enzymatically stable P2-purinoceptor agonist alpha,beta-methylene ATP increased the firing rate without altering the amplitude or shape of action potentials; the afterhyperpolarization following a spike was not changed either. When locus coeruleus neurons were hyperpolarized by current injection in order to prevent spontaneous firing, alpha,beta-methylene ATP produced depolarization and a slight increase in the apparent input resistance. A combined application of kynurenic acid and bicuculline methiodide failed to alter the alpha,beta-methylene ATP-induced depolarization, and tetrodotoxin only slightly depressed it. A gradual shift of the membrane potential by hyperpolarizing current injection led to a corresponding decrease, but no abolition or reversal of the alpha,beta-methylene ATP effect. In the hyperpolarized region, the current-voltage curve of alpha,beta-methylene ATP came into close approximation with, but did not cross, the control curve. Elevation of the external K+ concentration, or the intracellular application of Cs+ by diffusion from the microelectrode, depressed the response to alpha,beta-methylene ATP; external tetraethylammonium was also inhibitory. External Ba2+ and Cs+ had no effect or only slightly decreased the alpha,beta-methylene ATP-induced depolarization. A low Na+, or a low Ca2+ high Mg2+ medium, as well as the presence of Co2+ in the medium, markedly reduced or even abolished the depolarization by alpha,beta-methylene ATP. ATP itself did not produce consistent changes in the membrane potential or input resistance. However, in the presence of the P1-purinoceptor antagonist 8-cyclopentyl-1,3-dipropylxanthine, ATP consistently increased the firing rate and evoked an inward current. In conclusion, P2-purinoceptor activation appears to depolarize locus coeruleus neurons by inhibiting a persistent potassium current, and at the same time opening calcium-sensitive sodium channels or calcium-sensitive non-selective cationic channels.