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Hypotensive stress engages corticotropin-releasing factor (CRF) release within the rat locus coeruleus (LC), which activates LC neurones, initiating norepinephrine release in forebrain and activation of forebrain electroencephalographic activity. This study identified CRF afferents to the LC that are engaged during hypotensive stress. One of two potential CRF afferents, the central nucleus of the amygdala (CNA) or bed nucleus of the stria terminalis (BNST), was electrolytically lesioned and LC activation during hypotensive stress was quantified. Neither lesion altered LC spontaneous discharge rate or activation by intra-LC administered CRF. By contrast, LC activation by hypotensive stress was greatly attenuated in CNA-lesioned, but not BNST-lesioned, rats. Hypotensive stress-induced changes in transcriptional activation were immunohistochemically identified in CRF neurones that were retrogradely labelled from the LC region. c-fos immunoreactivity was prevalent in the paraventricular nucleus of the hypothalamus (PVN), CNA and BNST. However, only the PVN contained a substantial number of neurones that were doubly immunolabelled for CRF and c-fos, and few of these were retrogradely labelled from the LC. By contrast, immunoreactivity for the phosporylated form of cyclic AMP response-element binding protein (PCREB) was prevalent in CRF neurones in the CNA and BNST. Moreover, approximately one-third of the PCREB-expressing CRF neurones in the CNA were retrogradely labelled from the LC. These electrophysiological and anatomical findings implicate the CNA as a primary source of CRF that activates the LC during hypotensive stress. Additionally, CREB phosphorylation, rather than c-fos induction, is associated with hypotensive activation of CRF-CNA neurones that project to the LC.

Substantial clinical and preclinical findings support an association between stress and opiate abuse. To understand the mechanisms underlying this association, it is important to identify substrates of the stress response and endogenous opioid systems that interact and specific points at which stress circuits and endogenous opioid systems intersect.This review focuses on corticotropin-releasing factor (CRF), a critical substrate of the stress response, and its potential for interactions with endogenous opioid systems within the pontine nucleus, locus coeruleus (LC), a brain region that has been implicated as a target in response to stress and opiates.Evidence is reviewed supporting the hypothesis that CRF and endogenous opioids interact to co-regulate the LC. Thus, CRF- and enkephalin-immunoreactive fibers innervating LC dendritic fields overlap, and some axon terminals in this region co-localize CRF and enkephalin. CRF and opioids have opposing effects on LC neuronal discharge and on intracellular signaling mechanisms within LC neurons. Finally, a history of stress or opiate use induces plasticity in CRF-LC or opiate-LC interactions, respectively. Disruptions in the CRF/opioid balance as a result of this plasticity are proposed to result in hyperactivity or hyperresponsiveness of the LC-norepinephrine (NE) system.Co-regulation of the LC-NE system by CRF and opioids may be important in acute adaptation to stress. Potential clinical consequences of an imbalance in this regulation as a result of prior stress include increased risk of opiate self administration and decreased sensitivity to opiates used in clinical settings. Conversely, chronic exposure to opiates may predispose individuals to stress-related psychiatric disorders.

Neural circuits that are positioned to regulate rat distal colon function were identified by immunohistochemical detection of pseudorabies virus (PRV) and corticotropin-releasing factor (CRF). The distribution of PRV-immunoreactive neurons was examined in spinal cord and brain at increasing times (72-118 hours) after distal colon injection. At 72-80 hours, PRV-labeling was confined to the spinal cord, in the parasympathetic preganglionic column in the lumbosacral spinal cord and in the intermediolateral column of the thoracic spinal cord. At longer survival times (88 hours), PRV-immunolabeled neurons in the lumbosacral spinal cord were also distributed in superficial layers of the dorsal horn, the dorsal commissure, and around the central canal. Trans-synaptic labeling was identified in the medullary raphe nuclei, parapyramidal region, A5, Barrington's nucleus, A7, and the dorsal cap of the paraventricular nucleus of the hypothalamus after longer survival times (88-91 hours). Substantial labeling of the locus coeruleus, periaqueductal gray and forebrain regions occurred at later survival times (or = 96 hours). In dual-labeled sections, CRF terminal labeling surrounded PRV-labeled neurons in the parasympathetic preganglionic column of the lumbosacral spinal cord. Additionally, many neurons in Barrington's nucleus, but not other CRF-containing nuclei, were double labeled for CRF and PRV. These results, taken with previous studies, support a convergence in transneuronal labeling from different pelvic viscera that may be related to coordination of overall pelvic visceral functions. Importantly, they provide an anatomic substrate for an impact of CRF from Barrington's nucleus in normal and pathophysiological functions of the distal colon.

Neurochemical and electrophysiological studies indicate that the locus coeruleus (LC)-norepinephrine system is activated by physiological and external stressors. This activation is mediated in part by corticotropin-releasing factor (CRF), the hypothalamic neurohormone that initiates the endocrine response to stress. We have previously shown that the central nucleus of the amygdala (CNA) provides CRF afferents to noradrenergic processes in the peri-LC area that may serve to integrate emotional and cognitive responses to stress. The bed nucleus of the stria terminalis (BNST) shares many anatomical and neurochemical characteristics with the CNA, including a high density of CRF-immunoreactive cells and fibers; however, recent studies have suggested that the CNA and the BNST may differentially regulate responses to conditioned and unconditioned fear, respectively, suggesting divergent neuroanatomical circuits underlying these processes.In the present study, neuroanatomical substrates subserving regulation of the LC by the BNST were examined. Anterograde tract-tracing was combined with immunoelectron microscopy to test the hypotheses that BNST efferents target noradrenergic neurons of the LC and that these efferents exhibit immunolabeling for CRF.Ultrastructural analysis of sections that were dually labeled for the anterograde tracer biotinylated dextran amine (BDA) injected into the BNST and tyrosine hydroxylase (TH)-immunoreactivity demonstrated that BDA-labeled axon terminals formed synaptic specializations (primarily inhibitory) with TH-labeled dendrites and dendrites that lacked TH immunoreactivity. In contrast to CNA efferents that exhibited substantial immunolabeling for CRF, far fewer BDA-labeled terminals from the BNST in the rostrolateral peri-LC contained CRF.The present results indicate that the BNST may provide distinct neurochemical regulation of the peri-LC as compared to other limbic afferents such as the CNA. These data are interesting in light of behavioral studies showing that the CNA and BNST may be differentially involved in fear versus anxiety, respectively.

Corticotropin-releasing factor (CRF) acts as a putative neurotransmitter in the locus ceruleus (LC) to mediate its activation by certain stressors. In this study, we quantified LC sensitivity to CRF 24 h after swim stress, at a time when behavioral depression that is sensitive to antidepressants is apparent. Rats were placed in a tank with 30 cm (swim stress) or 4 cm water and 24 h later, either behavior was monitored in a forced swim test or LC discharge was recorded. Swim stress rats were more immobile than control animals in the swim test. LC neurons of swim stress rats were sensitized to low doses of CRF (0.1-0.3 microgram i.c.v.) that were ineffective in control animals and were desensitized to higher doses. Swim stress selectively altered LC sensitivity to CRF because neither LC spontaneous discharge nor responses to other agents (e.g., carbachol, vasoactive intestinal peptide) were altered. Finally, the mechanism for sensitization was localized to the LC because neuronal activation by low doses of CRF was prevented by the intracerulear administration of a CRF antagonist. CRF dose-response curves were consistent with a two-site model with similar dissociation constants under control conditions but divergent dissociation constants after swim stress. The results suggest that swim stress (and perhaps other stressors) functionally alters CRF receptors that have an impact on LC activity. Stress-induced regulation of LC sensitivity to CRF may underlie behavioral aspects of stress-related psychiatric disorders.

Corticotropin-releasing factor (CRF), the neurohormone that initiates the endocrine limb of the stress response via its actions on the anterior pituitary, also acts as a neurotransmitter in the noradrenergic locus coeruleus (LC) to activate this system during stress. Because the central nucleus of the amygdala contains numerous CRF-immunoreactive neurones, the present study examined whether CRF projections from the central nucleus of the amygdala target LC dendrites, thereby providing a mechanism for limbic-CRF modulation of brain noradrenergic activity. Retrograde tracers injected into the rostrolateral pericoerulear region, where CRF-immunoreactive fibres are dense, labelled numerous CRF-immunoreactive neurones in the central nucleus of the amygdala. Consistent with this, ultrastructural analysis of the rostrolateral pericoerulear region in sections that were dually labelled for an anterograde tracer (biotinylated dextran amine, BDA) injected into the central nucleus of the amygdala and CRF immunoreactivity revealed that a substantial percentage (35%) of amygdaloid axon terminals were CRF-immunoreactive. These terminals formed synaptic specializations with unlabelled dendrites that were more often of the asymmetric (excitatory) type. Additionally, ultrastructural analysis of sections that were dually labelled to visualize CRF-and tyrosine hydroxlase-immunoreactivity demonstrated synaptic specializations between CRF-immunoreactive terminals and LC dendrites in the rostrolateral peri-LC, which were also frequently asymmetric. Taken together with previous ultrastructural findings that LC dendrites in the rostrolateral pericoerulear region are targeted by anterogradely labelled terminals from the central nucleus of the amygdala, the present results implicate this nucleus as a source of CRF that can impact on LC activity via effects on dendrites in the rostrolateral pericoerulear region. This cellular substrate for amygdaloid-CRF modulation of brain noradrenergic activity may serve as a mechanism for the integration of emotional and cognitive responses to stress.

Corticotropin-releasing factor (CRF) administered intracerebroventricularly (i.c.v.) activates noradrenergic locus coeruleus (LC) neurons of halothane-anesthetized and unanesthetized rats. This study used a technique for microinfusing CRF into the LC from calibrated micropipettes to characterize and quantify the effects of locally administered CRF on LC discharge in halothane-anesthetized rats. CRF (3-100 ng) microinfusion into the LC increased discharge rate in a dose-dependent manner from 28 +/- 8 to 105 +/- 26% above preinfusion discharge rates. The CRF dose-response curve generated by local microinfusion was parallel to, and shifted approximately 200-fold to the left, of that generated by i.c.v. administration. Intracoerulear microinfusion of the CRF antagonist, [DPhe12,Nle(21,38),CalphaMeLeu37]r/hCRF(12-41), greatly attenuated LC activation produced by a maximally effective dose of i.c.v. administered CRF, suggesting that these effects are primarily due to actions within the LC. In rats in which both LC discharge rate and norepinephrine levels in prefrontal cortex were measured by in vivo microdialysis, CRF microinfused into the LC increased both endpoints. Finally, LC activation produced by CRF (60 ng) microinfusion into the LC was associated with cortical electroencephalographic activation. Taken together with previous anatomical and electrophysiological evidence for endogenous CRF interactions in the LC, our results support the hypothesis that CRF serves as an excitatory neurotransmitter in the LC, and suggest that its actions on LC neurons are translated to enhanced norepinephrine release and an impact on cortical targets.

Physiological and immunohistochemical studies have suggested that corticotropin-releasing factor (CRF), the hypophysiotropic peptide that initiates endocrine responses to stress, may serve as a neurotransmitter to activate noradrenergic neurons in the nucleus locus coeruleus (LC). We combined immunoperoxidase labeling for CRF and immunogold-silver localization of the catecholamine-synthesizing enzyme tyrosine hydroxylase (TH) in single sections through the rat LC to determine potential substrates for interactions between these two transmitters. Light microscopic analysis indicated that CRF processes are dense and highly varicose in the rostral LC region in the vicinity of noradrenergic dendrites. Electron microscopy of this rostral region revealed that immunoperoxidase labeling for CRF was mainly restricted to axons and axon terminals and was rarely seen in somata or dendrites. Axon terminals containing CRF immunoreactivity varied in size, content of synaptic vesicles, and formation of synaptic specializations. The postsynaptic targets of the CRF-labeled axon terminals consisted of both TH-labeled dendrites and dendrites lacking detectable TH-immunoreactivity. Of 113 CRF-immunoreactive axon terminals, approximately 70% were in direct contact with TH-labeled and unlabeled dendrites. Of the CRF-labeled axon terminals forming synapses with TH-labeled and unlabeled dendrites, they were either of the asymmetric (excitatory type; 19%) or symmetric (inhibitory type; 11%) variety or did not form identifiable contacts in the plane of section analyzed. Unlabeled axon terminals and glial processes were also commonly located adjacent to the plasma membranes of CRF-labeled axon terminals. These results provide the first direct ultrastructural evidence that axon terminals containing CRF-immunoreactivity 1) directly contact catecholamine-containing dendrites within the rostral pole of the LC, 2) may presynaptically modulate other afferents, and 3) are often enveloped by astrocytic processes.

The present study compared the effects of two analogs of corticotropin-releasing factor (CRF), [D-Phe12,Nle21,38, C alpha MeLeu37]CRF12-41 (D-PheCRF12-41) and alpha helical CRF9-41, as antagonists of CRF in in vivo and in vitro assays. In halothane-anesthetized rats, intracerebroventricular (i.c.v.) administration of both analogs inhibited the activation of locus coeruleus (LC) neuronal discharge produced by CRF (3.0 micrograms, i.c.v.). LC activation by hypotensive stress elicited by intravenous (i.v.) infusion of nitroprusside was antagonized by the same doses of the CRF antagonists that were effective in antagonizing CRF, suggesting that the receptors involved in LC activation by CRF and by hypotensive stress are similar. However, D-PheCRF12-41 was approximately 100 times more potent than alpha helical CRF9-41 when administered i.c.v. The IC50 values for D-PheCRF12-41 as an antagonist of CRF and of nitroprusside were 0.16 and 0.14 microgram, i.c.v., respectively. The IC50 values for alpha helical CRF9-41 as an antagonist of CRF and of nitroprusside were 18 and 27 micrograms, i.c.v., respectively. In contrast, D-PheCRF12-41 was only slightly more potent than alpha helical CRF9-41 in antagonizing CRF-stimulated cyclic AMP production in rat brain homogenates, with IC50s of 78 +/- 15 and 260 +/- 30 nM for D-PheCRF12-41 and alpha helical CRF9-41, respectively. Moreover, the antagonists had similar affinities for CRF binding sites in rat brain homogenates, with Kis of 15.5 +/- 4 nM and 10.3 +/- 6 nM for D-PheCRF12-41 and alpha helical CRF9-41, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

The effects of corticotropin-releasing factor (CRF), a benzodiazepine inverse agonist (methyl-6,7-dimethoxy-4-ethyl-beta-carboline-3-carboxylate; DMCM) and electric foot-shock on rat conflict behavior were characterized and compared. Rats were trained to lever press under a multiple fixed-ratio schedule (FR 20) of food reinforcement in which responses during the first component were not punished, and the first response of each FR during the second component produced electric shock of an intensity sufficient to suppress responding by 10% to 15%. Intracerebroventricular injection of CRF (0.1-5.6 micrograms) caused a dose-dependent decrease in the rate of responding in both components of the schedule. However, CRF was more potent in decreasing rates of punished responding (proconflict effect). DMCM (10-100 micrograms; i.c.v.) also decreased rates of punished and nonpunished responding and was more potent during the punishment component. The suppression of punished and nonpunished responding by CRF and DMCM was mimicked by increasing the shock intensity (delta = 0.1 to 0.6 mA) during the punishment component. To determine whether CRF, DMCM and electric shock shared common mechanisms for these effects, rats were pretreated with i.c.v. injections of either a CRF antagonist (alpha helical CRF9-41, 50 micrograms), a benzodiazepine agonist (chlordiazepoxide, 10 micrograms) or a benzodiazepine antagonist (flumazenil, 10 micrograms) before the administration of equieffective doses of CRF or DMCM or an increase in shock intensity. Chlordiazepoxide attenuated the effects of all three stimuli. Flumazenil antagonized DMCM and CRF, but not shock, implicating a pharmacologic interaction between CRF and benzodiazepine systems.(ABSTRACT TRUNCATED AT 250 WORDS)

Corticotropin-releasing factor (CRF) has been implicated as a neurotransmitter in the noradrenergic nucleus, locus coeruleus (LC), and is thought to be hypersecreted in depression. Therefore, the hypothesis that antidepressants interfere with CRF neurotransmission in the LC was tested. The acute and chronic effects of desmethylimipramine (DMI), sertraline (SER), phenelzine (PHE), mianserin (MIA), and cocaine (COC) were quantified on LC spontaneous discharge, LC sensory-evoked discharge, LC activation by intracerebroventricular (i.c.v.)-administered CRF, and LC activation by stress in halothane-anesthetized rats. No consistent effect of the drugs on LC spontaneous discharge rate or sensory responsiveness was observed after acute administration. LC spontaneous discharge rates in rats chronically administered the drugs were similar to rates recorded in matched controls, with the exception of PHE and COC. In these rats, LC spontaneous discharge rates were lower than those of untreated rats. Interestingly, LC responses to repeated sciatic nerve stimulation (as measured by the ratio of evoked-to-tonic LC discharge rate) were enhanced in rats chronically administered SER and PHE. This is opposite to the reported effects of i.c.v.-administered CRF. This was not observed with chronic administration of DMI, MIA, or COC. The most striking effect associated with chronic administration of DMI and MIA was the attenuation of LC activation by hemodynamic stress. Because this activation requires CRF release in the LC region, and because none of the antidepressants altered LC activation by i.c.v.-administered CRF, the data suggest that chronic administration of either antidepressant, DMI or MIA, results in attenuation of stress-elicited CRF release in the LC.(ABSTRACT TRUNCATED AT 250 WORDS)

Neuroendocrine and catecholamine dysfunctions in depression may be linked by corticotropin-releasing factor (CRF) effects on locus coeruleus (LC) neurons. One consequence of CRF hypersecretion in depression would be persistent elevated levels of LC discharge and diminished responses to phasic sensory stimuli. The hypothesis that antidepressants could reverse these changes was tested by characterizing effects of pharmacologically distinct antidepressants on LC sensory-evoked discharge, LC activation by stress, and LC activation by CRF. The most consistent effect of all of the antidepressants tested was a decrease in LC sensory-evoked discharge after acute administration. However, tolerance occurs to these effects after chronic administration. With chronic administration each of the antidepressants produced effects which could potentially interfere with CRF function in the LC. Desmethylimipramine and mianserin attenuated LC activation by a stressor which requires endogenous CRF, suggesting that these antidepressants attenuate stress-elicited release of CRF and perhaps the hypersecretion that occurs in depression. The serotonin reuptake inhibitor, sertraline (SER), enhanced the signal-to-noise ratio of the LC sensory response, an effect opposite to that of CRF. Thus, SER could serve as a functional antagonist of CRF that is hypersecreted in depression. The finding that three pharmacologically distinct antidepressants share the potential to interfere with CRF function in the LC implies that this may be an important common mechanism for antidepressant activity.

Corticotropin-releasing factor (CRF), the hormone responsible for adrenocorticotropin release during stress, is thought to be hypersecreted in depression. Because recent studies suggest that CRF may serve as a neurotransmitter in the major noradrenergic nucleus, locus coeruleus (LC), it was hypothesized that antidepressants interfere with the putative neurotransmitter role of CRF in the LC by either: 1) decreasing release of CRF; 2) pharmacologically antagonizing CRF; or 3) functionally antagonizing CRF by producing effects on LC cells that oppose these of CRF. In order to test this hypothesis, the effects of acute and chronic administration of two antidepressants, a norepinephrine re-uptake inhibitor (desmethylimipramine, DMI) and a serotonin re-uptake inhibitor (sertraline, SER), on LC spontaneous discharge, LC sensory evoked discharge, LC activation by a stressor and LC activation by CRF, were compared in halothane-anesthetized rats. Acute i.v. administration of DMI decreased both LC spontaneous discharge and discharge evoked by repeated sciatic nerve stimulation. In contrast, acute i.v. SER administration decreased only evoked LC discharge rate. Chronic DMI administration (10.0 mg/kg/day, i.p., 21 days) resulted in tolerance to its effects on spontaneous and sensory-evoked LC discharge. However, chronic DMI administration attenuated LC activation by hemodynamic stress, which is thought to require CRF release. LC activation by intracerebroventricular CRF was not altered in the chronic DMI rats. In contrast to DMI, chronic SER (10 mg/kg/day, i.p., 21 days) did not alter LC activation by either stress of CRF. However, the response of LC cells to repeated sciatic nerve stimulation was somewhat enhanced in chronic SER rats. This is an effect that is opposite that produced by CRF.(ABSTRACT TRUNCATED AT 250 WORDS)

The effects of two benzazocines, UM-1037 and UM-1046, which produce a quasi-morphine abstinence syndrome in normal rhesus monkeys, were characterized by the use of the isolated guinea-pig ileal preparation and by a drug discrimination procedure in pigeons. These benzazocines produce a contraction of the ileum isolated from narcotic-naive guinea pigs similar to the narcotic antagonist-precipitated contraction of ilea isolated from morphine-pretreated animals. This effect is suppressed by morphine, and the suppression is reversed by naloxone. These benzazocines appear to stimulate the release of acetylcholine from ilea that have not been exposed to narcotics, and thus produce an effect similar to the release of acetylcholine produced by narcotic antagonists in ilea that have been exposed to morphine-like narcotics. The drug discrimination studies are also consistent with the involvement of cholinergic mechanisms in the behavioral effects produced by the benzazocines. In pigeons that have been trained to discriminate UM-1046 from saline, physostigmine, but not neostigmine, produces responses like those produced by UM-1046. In contrast, drugs from other pharmacological classes do not share discriminative stimulus properties with UM-1046. Thus, complementary evidence was obtained from both a physiological and a behavioral approach. The involvement of cholinergic pathways in the quasi-abstinence syndrome produced by UM-1037 and UM-1046 suggests that similar pathways might be active during narcotic abstinence.

In order to determine whether pharmacologically-induced alterations in the spontaneous activity of neurons in the locus coeruleus are associated with changes in blood pressure, the activity of the locus coeruleus and blood pressure were recorded simultaneously in anesthetized rats after the administration of agents known to affect both of these parameters. Spontaneous activity of the locus coeruleus was decreased by intracerebroventricular (i.c.v.) administration of both clonidine and St 91, [2,(2,6-diethyl-phenylimino)imidazolidine chloride], a charged analogue of clonidine. However, only clonidine decreased the mean blood pressure after intracerebroventricular administration suggesting that either the receptors mediating decreases in the activity of the locus coeruleus are different to those mediating hypotension, or that St 91 does not distribute to sites involved in the control of blood pressure even after intracerebroventricular administration. Intravenous administration of clonidine, but not of St 91, decreased the activity of the locus coeruleus and produced a prolonged hypotension, thus suggesting a central mechanism for these effects. Both clonidine and St 91 administered intravenously, produced a brief initial period of hypertension which was not associated with consistent changes in the spontaneous activity of the locus coeruleus. Thus, noradrenergic agonists can decrease the activity of the locus coeruleus without affecting blood pressure, and increase blood pressure without affecting the activity of the locus coeruleus. The spontaneous activity of cells in the locus coeruleus was increased by 100% after the intracerebroventricular administration of corticotropin-releasing factor (CRF; 3.0 micrograms).(ABSTRACT TRUNCATED AT 250 WORDS)

The effects of dynorphin-A (Dyn) and [Leu5]-enkephalin (Enk) were compared in the vitro hippocampal slice preparation, using extracellular field potential and intracellular voltage recordings. In the CA1 region, Dyn, like Enk, consistently increased the size of the extracellularly recorded population spike (PS) evoked by stratum radiatum (StR) stimulation of the Schaffer collaterals. These responses were naloxone reversible. In contrast, in the CA3 region, Dyn both increased and decreased the PSs evoked by mossy fiber stimulation, whereas Enk slightly enhanced the PS. Intracellular recordings from CA3 pyramidal neurons (HPNs) revealed both excitatory and inhibitory actions of Dyn on spontaneous activity. Associated membrane potential changes were variable. In contrast, Enk had only weak effects on spontaneous activity and no effect on membrane potential. These data suggest regional differences in the effect of Dyn and Enk on hippocampal activity.

The effects of hemodynamic stress produced by i.v. infusion of nitroprusside on discharge characteristics of noradrenergic locus ceruleus (LC) neurons were compared to those produced by central administration of corticotropin-releasing factor (CRF). Like CRF, both nitroprusside infusion and blood volume withdrawal increased LC spontaneous discharge. Additionally, nitroprusside disrupted the response of LC cells to repeated sciatic nerve stimulation in a manner similar to CRF. The neuronal effects of nitroprusside were prevented by prior administration of a CRF antagonist, alpha helical CRF9-41, but not by dexamethasone, which blocks CRF release into the hypophysial portal circulation. These results suggest that LC activation by hemodynamic stress is mediated by neuronal CRF release onto LC neurons. alpha Helical CRF9-41 had no effect on the magnitude or timecourse of nitroprusside-elicited hypotension, suggesting that LC activation by nitroprusside is not important for maintaining hemodynamic homeostasis. Finally, chronic administration of desmethylimipramine (DMI) attenuated the response of LC cells to nitroprusside, but not to CRF, suggesting that antidepressant therapy may inhibit stress-induced CRF release. This study supports the concept that CRF serves as a neurotransmitter in the LC in the initiation of stress responses.

Because the physiological characteristics of noradrenergic locus ceruleus (LC) neurons differ in anesthetized and unanesthetized rats, the effects of morphine on LC activity recorded in both conditions were compared. Intracerebroventricular administration of morphine inhibited spontaneous LC discharge of both anesthetized and unanesthetized rats but morphine was at least 10 times more potent in anesthetized rats. In anesthetized rats LC discharge evoked by sciatic nerve stimulation was insensitive to doses of morphine (0.03 or 0.1 microgram) that inhibited LC spontaneous activity. Only the highest dose of morphine (0.3 microgram) which inhibited completely tonic activity decreased significantly evoked discharge. In parallel experiments in unanesthetized rats, the presentation of auditory stimuli evoked a pattern of LC discharge similar to that evoked by sciatic nerve stimulation in anesthetized rats. Morphine (1.0 and 3.0 micrograms) decreased both spontaneous and evoked activity in these rats; however, spontaneous LC discharge was more sensitive to morphine, as was observed with anesthetized rats. Quantitative analyses of these effects indicated that morphine tends to alter the pattern of LC discharge to sensory stimuli such that the signal-to-noise ratio (ratio of evoked/tonic activity during stimulus presentation) is increased. Morphine effects were reversed by naltrexone (1.0 mg/kg s.c.) in anesthetized rats and 1.0 microgram i.c.v. in unanesthetized rats. The present results indicate that the degree of arousal and the availability of environmental stimuli are important determinants of opiate effects on LC activity.