Electrophysiological correlates of presynaptic opiate receptor activation: reduction in norepinephrine-mediated inhibition from the locus coeruleus

Inhibitory responses of rat cerebellar Purkinje cells to locus coeruleus (LC) stimulation and iontophoresis of norepinephrine (NE) were examined before and after administration of morphine to determine whether the inhibitory modulation of NE release by opiates results in a functional impairment in noradrenergic synaptic action. Administration of morphine systematically (0.2-1.2 mg/kg, i.v.) or by iontophoresis reduced inhibitions in Purkinje firing elicited by LC stimulation without affecting depressions in activity induced by postsynaptic applications of NE. This antagonistic effect of morphine on LC-induced inhibition was reversed or prevented by naloxone and mimicked by administration of levorphanol but not dextrorphan. Morphine increased the excitatory response of Purkinje cells to monosynaptic input from the parallel fibers, whereas it blocked [gamma]-aminobutyric acid-induced inhibitions in firing via a non-opiate receptor-mediated mechanism. These results demonstrate that morphine interferes with synaptic inhibition derived from the LC and suggest that this may occur via activation of presynaptic opiate receptors residing on noradrenergic nerve terminals.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/26539/1/0000078.pd

Inhibitory responses of rat basolateral amygdaloid neurons recorded in vitro

The purpose of the present study was to characterize the ionic and pharmacological basis of the actions of synaptically released and exogenously applied GABA in basolateral amygdaloid pyramidal cells in vitro. Stimulation of forebrain afferents to pyramidal neurons in the basolateral amygdala evoked an excitatory postsynaptic potential followed by early and late inhibitory postsynaptic potentials. The early inhibitory postsynaptic potential had a reversal potential near -70 mV, was sensitive to changes in the chloride gradient across the membrane and was blocked by the GABAA antagonists picrotoxin and bicuculline methiodide but not by the GABAB antagonists phaclofen or 2-hydroxysaclofen. In contrast, the late inhibitory postsynaptic potential had a reversal potential of approximately -95 mV and was markedly reduced or abolished by GABAB antagonists.Pressure application of GABA to the surface of the slice typically elicited a triphasic response in basolateral amygdaloid pyramidal neurons consisting of a short-latency hyperpolarization that preceded or was superimposed on a membrane depolarization followed by a longer latency hyperpolarization. Each of the responses was associated with an increase in membrane conductance. Determinations of the reversal potential, ionic dependency and sensitivity to pharmacological blockade of each component of the GABA-induced response revealed that the initial hyperpolarizing (Erev approximately -70 mV) and depolarizing (Erev approximately -55 mV) responses were mediated by a GABAA-mediated increase in chloride conductance, whereas the late hyperpolarizing response (Erev approximately -82 mV) to GABA arose from a GABAB-mediated increase in potassium conductance. Experiments in which GABA was applied at various locations on the cell suggested that the short-latency hyperpolarization resulted from activation of somatic GABA receptors, whereas the depolarizing and late hyperpolarizing responses were generated primarily in the dendrites. In contrast to the complex membrane response profile elicited by GABA, pressure ejection of the GABAB agonist baclofen produced only membrane hyperpolarizations.Taken together, these results suggest that inhibitory responses that are recorded in basolateral amygdaloid pyramidal cells are mediated by activation of both GABAA and GABAB receptors. Consistent with findings elsewhere in the CNS, the early inhibitory postsynaptic potential and initial hyperpolarization and depolarizing response to local GABA application appear to involve a GABAA-mediated increase in chloride conductance, whereas the late inhibitory postsynaptic potential and the late hyperpolarizing response to GABA arise from a GABAB-mediated increase in potassium conductance.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/29831/1/0000178.pd

Electrophysiological effects of dynorphin peptides on hippocampal pyramidal cells in rat

Single-unit extracellular recording was carried out in rats to characterize the effects of dynorphin and several structurally related peptides on hippocampal pyramidal cell activity. Dynorphin, applied electrophoretically or by pneumatic pressure, produced a dose-dependent depression of both spontaneous and glutamate-evoked discharge in a majority (63%) of CA1 and CA3 cells tested. In addition, a small number of cells in both cellular fields responded to the peptide with a prolonged elevation in firing. The inhibitory effects of dynorphin in were not blocked by naloxone. Moreover, administration of des-tyrosine-dynorphin depressed the firing of pyramidal cells in a manner similar to that of the parent compound. Ethylketocyclazocine produced a mixed pattern of excitatory and inhibitory effects, whereas naloxone-sensitive elevations in firing were most often observed with the application of dynorphin-(1-8). Application of [Leu5]enkephalin produced only facilitations in pyramidal cell firing. The possibility is raised that biologically significant non-opiate actions, in addition to potent opiate-mediated effects, may occur upon release of pro-dynorphin peptides in the hippocampus.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/25783/1/0000344.pd

Changes occur in central adrenoreceptor function following long-term morphine treatment and during morphine withdrawal

Radioloigand binding techniques were used in combination with electrophysiological recording to characterize changes in beta adrenoreceptor activity in various brain areas in rats treated chronically with morphine. Following chronic morphine treatment, the maximum number of specific binding sites for 3H-dihydroalprenolol (3H-DHA) in parietal cortex and hippocampus showed a biphasic change, indicating an initial increase and decrease (relative to controls) in beta adrenoreceptors in these regions with time during withdrawal. No appreciable changes were observed in the dissocation constants for 3H-DHA binding. The changes in cortical beta adrenoreceptor density found in early (8 hr) and later phases (32 hr) of withdrawal were paralled by a selective increase and decrease, respectively, in cortical neuron sensitivity to noradrenergic stimulation. These results suggest a possible linkage between changes in central adrenoreceptor function and the formation and/or expression of opiate dependence.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/24607/1/0000017.pd

Muscarinic modulation of conductances underlying the afterhyperpolarization in neurons of the rat basolateral amygdala

The excitability level of pyramidal neurons in the basolateral amygdala (BLA) is greatly increased following muscarinic receptor activation, an effect associated with an increased rate of action potential firing and reduction of the afterhyperpolarization (AHP). We impaled BLA pyramidal neurons in slices of rat ventral forebrain with a single microelectrode to examine the currents underlying the AHP and spike frequency accomodation and determine their sensitivities to muscarinic modulation. In voltage-clamp, depolarizing steps were followed by biphasic outward tail currents, consisting of rapidly decaying (IFast) and slowly decaying (ISlow) current components. These corresponded temporally with the medium and slow portions of the AHP, respectively. The reversal potential or the IFast component of the AHP tail current shifted in the depolarizing direction with increases in the extracellular K+ concentration. The amplitude of IFast was reduced during perfusion of 0-Ca2+ medium or by superfusion of TEA (1-5 mM) or carbachol (10-40 [mu]M). It is suggested that IFast was produced by the rapidly decaying Ca2+-activated K+ current (IC) and the muscarinic-sensitive M-current (IM). The ISlow tail current component reversed at the estimated values for EK in medium containing either normal or elevated K+ levels. This component was eliminated by perfusion of 0-Ca2+ medium or inclusion of cyclic-AMP in the recording electrode. It was not blocked by TEA (5 mM) or apamin (50-500 nM), but was reduced by carbachol in a dose-dependent manner (IC50=0.5 [mu]M). Electrical stimulation cholinergic afferent pathways to the BLA produced inhibition of ISlow, an effect which was enhanced by eserine and prevented by atropine. Loss of the ISlow component was always accompanied by similar reductions in accomodation and the slow AHP. It was concluded that this tail current component resulted from the slowly decaying Ca2+-activated K+ current, IAHP. Thus, the muscarinic inhibition of IAHP contributes to the enhanced excitability exhibited by BLA pyramidal neurons following cholinergic stimulation.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/30577/1/0000212.pd

Adrenergic responses of baroreceptive cells in the nucleus tractus solitarii of the rat: a microiontophoretic study

Anatomical and pharmacological evidence suggests a role for catecholamines (CAs) in the modulation of the baroreceptor reflex within the nucleus tractus solitarii (NTS). Single neurons in the NTS of the rat were studied for their responses to activation of the baroreceptor reflex and to iontophoretic administration of dopamine, norepinephrine (NE), and epinerphrine (EPI) to determine the relationship between the effects of baroreflex activation and CA application on baroreceptive neurons in the vagal sensory nucleus. Of 269 cells studied, 104 (38.7%) exhibited decreases and 41 cells (15.2%) showed increases in firing rate in response to baroreflex activation, while the remaining 124 neurons showed no response. All 3 CAs inhibited spike activity in the majority (68.5%) of NTS cells. These inhibitory effects on spontaneous firing were observed regardless of the response profile of a particular neuron to baroreflex activation. The inhibitory effects of NE and EPI on NTS neuronal activity were specifically blocked by the [alpha]-adrenergic receptor antagonist tolazoline, but not by the [beta]-adrenergic antagonist sotalol. These results indicate that CAs may interact at several sites within the NTS to influence baroreflex integration, and that the effects of NE and EPI on neuronal activity are mediated by an [alpha]-adrenergic receptor.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/26574/1/0000113.pd

Ca 2+ and frequency dependence of exocytosis in isolated somata of magnocellular supraoptic neurones of the rat hypothalamus

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/66206/1/jphysiol.2003.051136.pd

Multiple effects of long-term morphine treatment on postsynaptic [beta]-adrenergic receptor function in hippocampus: an intracellular analysis

We previously reported that [beta]-adrenergic receptors are increased in cerebral cortex and hippocampus in rats treated chronically with morphine and subsequently down-regulated after morphine withdrawal [22,23]. The changes in receptor density in hippocampus were accompanied by a corresponding super- and subsensitivity, respectively, in [beta]-adrenergic responsiveness, as assessed electrophysiologically by measuring the ability of isoproterenol to augment population spike responses in the slice. In this study, we compared the ability of isoproterenol to reduce the Ca2+-activated K+ slow afterhyperpolarization (slow AHP) in pyramidal neurons in hippocampal slices from opiate-naive and chronic morphine-treated rats to determine whether such changes in [beta]-adrenergic receptor function are localized postsynaptically. Chronic treatment of rats with morphine produced a 3.5-fold parallel shift to the left in the concentration-response curve for isoproterenol and reduced the EC50 from 4.8 +/- 1.3 to 1.4 +/- 0.5 nM. In contrast, sensitivity and maximal responsiveness to isoproterenol was markedly decreased in pyramidal neurons recorded in slices from morphine withdrawn animals. The concentration-response curves for inhibition of the slow AHP by carbachol or forskolin were not affected by chronic morphine treatment. However, blockade of the slow AHP by forskolin was significantly reduced in pyramidal neurons studied after morphine withdrawal. These data suggest that the increase in electrophysiological responsiveness to [beta]-adrenergic receptor stimulation found in hippocampus after chronic morphine treatment most likely resulted from an up-regulation in postsynaptic membrane receptors, whereas alterations occurring beyond the receptor level may be involved in the desentization that is associated with morphine withdrawal.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/31332/1/0000241.pd

Modification of the visual response properties of cerebellar neurons by norepinephrine

Extracellular recordings were conducted in the paraflocculus of anesthetized Long-Evans pigmented rats to determine how iontophoresis of norepinephrine (NE) affects the responsiveness of individual Purkinje cells and interneurons to presentations of visual stimuli within their visual receptive fields. Presentations of moving or stationary visual stimuli during the control (pre-NE) period elicited simple spike excitations or inhibitory responses in slightly more than one-half (55%, n = 32) of the cells tested (20 of 38 Purkinje cells, 12 of 20 interneurons). The predominant effect of NE iontophoresis was to improve visually evoked responses in those neurons which showed modulations in their simple spike discharge to control presentations of visual stimuli. A clear enhancement of visual responses by NE (i.e., absolute increase over control) was observed in 18 of the units, and in 12 of the 14 remaining cells, reductions in stimulus-bound discharge during catecholamine iontophoresis were accompanied by much larger depressions in background activity, resulting in a net enhancement in the ratio of signal-to-noise. NE differentially affected responses to stimulus movement in the preferred and non-preferred direction in one-third of these neurons, such that directional selectivity was increased. However, the orientation bias of individual units was unchanged by NE. Iontophoretic application of the [beta]-adrenergic antagonist sotalol but not the [alpha]-adrenergic antagonist phentolamine blocked these facilitating noradrenergic effects. An additional feature of noradrenergic action was revealed in tests conducted in 26 cells which did not respond to control presentations of visual stimuli. Iontophoresis of NE resulted in the elicitation of visual responses in 11 of these units, suggesting the possibility that NE might act in some cases to gate the efficacy of subliminal synaptic input conveyed by classical afferent channels. It is proposed that an important aspect of noradrenergic action within local cerebellar circuits might be to refine the receptive field properties of individual neuronal elements and thereby improve information flow through the cerebellum.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/28612/1/0000424.pd

Electrophysiological actions of VIP in rat somatosensory cortex

Electrophysiological and biochemical studies suggest that VIP may exert a facilitating action in the neocortical local circuitry. In the present study, we examined the actions of VIP and VIP + norepinephrine (NE) on somatosensory cortical neuron responses to direct application of the putative transmitters acetylcholine (ACh) and gamma-aminobutyric acid (GABA). Spontaneous and transmitter-induced discharges of cortical neurons from halothane-anesthetized rats were monitored before, during and after VIP, NE and VIP + NE iontophoresis. In 57 VIP-sensitive cells tested, VIP application (5-70 nA) increased (n = 18), decreased (n = 36) or had biphasic actions (n = 3) on background firing rate. In a group of 20 neurons tested for NE + VIP, the combined effect of both peptide and bioamine was predominantly (70%) inhibitory. On the other hand, inhibitory and excitatory responses of cortical neurons to GABA (11 of 15 cases) and ACh (10 of 18 cases), respectively, were enhanced during VIP iontophoresis. Concomitant application of VIP and NE produced additive (n = 2) or more than additive (n = 3) enhancing effects on GABA inhibition. NE administration reversed or enhanced further VIP modulatory actions on ACh-induced excitation. These findings provide electrophysiological evidence that NE and VIP afferents may exert convergent influences on cortical neuronal responses to afferent synaptic inputs such that modulatory actions are anatomically focused within the cortex.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/29264/1/0000322.pd