Organization of projections from the medial agranular cortex to the superior colliculus in the rat: a study using anterograde and retrograde tracing methods

The organization of corticotectal projections from the medial agranular cortex (AGm), which has been considered to contain rat's frontal eye field, was examined using anterograde and retrograde tracing techniques. When biotinylated dextranamine (BDA) injections were made into the rostral part of the AGm, small numbers of BDA-labeled axons were found in the rostral two-thirds of the superior colliculus (SC) while some labeled axons were seen in the caudal one-third of the SC. These labeled axons were distributed mainly in the lateral part of the stratum griseum intermediale. On the other hand, after BDA injections into the caudal part of the AGm, moderate to dense plexuses of labeled axons were found in the rostral two-thirds of the SC while some labeled axons were seen in the caudal one-third of the SC. These labeled axons were distributed in the ventromedial and dorsolateral marginal zones of the stratum griseum intermediale as well as in the stratum griseum profundum. The corticotectal projections were largely uncrossed. After combined injections of BDA into the caudal part of the AGm on one side and cholera toxin B subunit (CTb) into the paramedian pontine reticular formation on the opposite side or into the interstitial nucleus of Cajal on the same side, the overlapping distributions of BDA-labeled axons and CTb-labeled neurons were found in the ventromedial marginal zone of the stratum griseum intermediale ipsilateral to the site of BDA injection. These results suggest that the caudal part of the AGm plays a more significant role in the oculomotor function than does the rostral part of the AGm.     

Effects of physiological manipulations on locus coeruleus neuronal activity in freely moving cats. II. Cardiovascular challenge

Several cardiovascular manipulations were examined for their effects on single-unit activity of locus coeruleus noradrenergic (LC-NE) neurons in unanesthetized, unrestrained cats: hydralazine (1 mg/kg, i.v.) was administered to present a tonic hypotensive stimulus, and to activate preferentially the neural component of the sympathoadrenal system; hemorrhage was used to decrease blood volume and to activate both the neural and hormonal components of the sympathoadrenal system; intravenous infusion of isotonic saline was used to increase blood volume. LC-NE neurons were activated by hydralazine, in parallel with the sympathetic response (indicated by elevated heart rate and plasma NE). LC-NE unit activity was decreased following a volume load. However, contrary to previous findings in anesthetized animals, hemorrhage had no effect on LC-NE unit activity, but did activate both components of the sympathetic response. It is concluded that: (1) cardiovascular stimuli can influence the activity of LC-NE neurons, though they show less sensitivity to such stimuli than do primary regulatory mechanisms; (2) the response of LC-NE neurons to physiological stimuli can occur independent of changes in behavioral state; (3) these neurons do not appear to play a specific role in cardiovascular regulation, but may respond to physiological challenges in general; (4) finally, in agreement with previous studies, our data show that LC-NE neurons are generally co-activated with the sympathetic nervous system, but also that the two can be dissociated (e.g. hemorrhage).     

The release of noradrenaline in the locus coeruleus and prefrontal cortex studied with dual-probe microdialysis

The present study was undertaken to investigate and compare the properties of noradrenaline release in the locus coeruleus (LC) and prefrontal cortex (PFC). For that aim the dual-probe microdialysis technique was applied for simultaneous detection of noradrenaline levels in the LC and PFC in conscious rats. Calcium omission in the LC decreased noradrenaline levels in the LC, but increased its levels in the PFC. Novelty increased noradrenaline levels in both structures. Infusion of the alpha(2)-adrenoceptor agonist clonidine decreased extracellular noradrenaline in the LC as well as in the PFC. Infusion of the alpha(2A)-adrenoceptor antagonist BRL44408, or the alpha(1)-adrenoceptor agonist cirazoline into the LC or PFC caused a similar dose-dependent increase in both structures. When BRL44408 or cirazoline were infused into the LC, few effects were seen in the PFC. Infusion of the 5-HT(1A)-receptor agonist flesinoxan into the LC or the PFC decreased the release of noradrenaline in both structures. When flesinoxan was infused into the LC, no effects were seen in the PFC. When the GABA(A) antagonist bicuculline was applied to the LC, noradrenaline increased in the LC as well as in the PFC. It is concluded that the release of noradrenaline from somatodendritic sites and nerve terminals responded in a similar manner to presynaptic receptor modulation. The possible existence of dendritic noradrenaline release is discussed.     

The corticocortical projections of the physiologically defined eye field in the rat medial frontal cortex

This study investigated in the rat the corticocortical projections of the frontal eye field (FEF), which is located within the medial frontal cortex. The experiments were carried out on Wistar rats. Seven animals received a single iontophoretic injection of Phaseolus vulgaris leucoagglutinin in an FEF site within the medial frontal cortex where intracortical microstimulation elicited eye movements. In these cases, anterogradely labeled fibers and terminal-like elements were found in both hemispheres. The densest labeling was seen in the injected hemisphere, where labeled fibers prevailed in the visual cortex and their laminar distribution differed between the primary and secondary visual cortices. Dense labeled fibers were also seen in the frontal and retrosplenial cortex, whereas a columnar arrangement of terminal-like elements was detected in a restricted part of area 1 of the somatosensory cortex. Contralaterally to the injection site, labeled fibers were distributed mainly in the homotopic region. In two animals, the tracer was injected in a site at the FEF border whose stimulation evoked eye and whisker movements. In these animals, a different distribution of labeling was observed with respect to the other rats in which the tracer was deposited within the FEF, and anterograde labeling was observed in areas 1 and 2 of the parietal cortex of both hemispheres; in addition, no labeling was observed in these cases in the primary visual cortex. These findings suggest that cortical sites confined within the rat FEF are implicated in the control of orienting and exploring behaviors in addition to the control of eye movement.     

Stimulation of the locus coeruleus suppresses trigeminal sensorimotor function in the rat

The nucleus locus coeruleus (LC) has been implicated in the modulation of the spinal sensorimotor function. The aim of the present study was to examine the effect of electrical stimulation of the LC on sensorimotor function in the trigeminal system. The following two cases of sensorimotor behaviors mediated by the trigeminal brainstem sensory nuclear complex were examined: (1) the activity of the masseter muscle evoked by pressure on the region of the temporomandibular joint (TMJ); and (2) the activity of the digastric muscle evoked by electrical stimulation of the tooth pulp, resulting in the jaw-opening reflex. In the first case, LC stimulation at 10, 30 and 50 microA resulted in a 70%, 68% and 55% reduction in the magnitude of electromyogram (EMG) activity of the masseter muscle compared with the control (without LC stimulation), respectively. The threshold intensity for the onset of masseter EMG activity increaced to 106%, 111% and 121% of the control with 10, 30 and 50 microA LC stimulation, respectively. In the second case, EMG magnitude in response to the digastric muscle decreased to 42% of the control when 30 microA of LC stimulation was delivered. These results suggest that descending influences from the LC can act in suppression of the trigeminal sensorimotor function.     

Effects of acute and chronic clozapine on dopaminergic function in medial prefrontal cortex of awake, freely moving rats

We previously showed that chronic administration of the clinically atypical and clinically superior antipsychotic drug clozapine selectively reduces dopamine (DA) release in the nucleus accumbens but not neostriatum, and that this effect appears mediated by anatomically selective mesolimbic DA depolarization blockade. The present study extends that research to another mesocorticolimbic DA locus, the medial prefrontal cortex. Acute clozapine challenge (5-40 mg/kg i.p.) produced dose-dependent increased extracellular levels of DA and its metabolites, 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA), in the medial prefrontal cortex of awake, free-moving rats as measured by in vivo brain microdialysis. Chronic clozapine treatment (20 mg/kg/day for 21 days) did not significantly change basal extracellular levels of DA, DOPAC or HVA. Acute clozapine challenge on day 22 in the chronic clozapine-treated animals produced no significant differences in medial prefrontal cortex DA, DOPAC or HVA as compared to chronic vehicle-treated animals, indicating that tolerance to clozapine does not develop in the mesocortical DA system, in contrast to the mesolimbic system. The DA agonist apomorphine (100 micrograms/kg) produced decreased basal extracellular levels of DA, DOPAC and HVA in medial prefrontal cortex of both chronic clozapine-treated and chronic vehicle-treated rats.(ABSTRACT TRUNCATED AT 250 WORDS)     

Single unit activity of noradrenergic neurons in locus coeruleus and serotonergic neurons in the nucleus raphe dorsalis of freely moving cats in relation to the cardiac cycle

Single unit activity of serotonergic (5-HT) neurons in the nucleus raphe dorsalis (NRD) and of noradrenergic (NE) neurons in the locus coeruleus (LC) was recorded in relation to the cardiac cycle in awake, freely moving cats. The discharge of NRD-5-HT neurons showed no relationship to the cardiac cycle, while LC-NE neurons displayed a cardiac periodicity such that the units were most likely to fire from 80 to 180 ms after the peak of the cardiac r-wave (diastole), and least likely to fire during the period from 40 ms before to 60 ms after the r-wave (systole). The strength of this periodicity was inversely related to the discharge rate of individual cells. Exposure to a noxious environmental stimulus (15 min of 100 dB white noise) greatly attenuated the cardiac relationship of LC-NE neurons. A blood volume expansion of approximately 15% (9.0 ml/kg b. wt.) decreased unit rate by about 25%, but did not alter either the timing or the magnitude of the LC-NE cardiac relationship. These data are discussed in terms of the participation of NRD-5-HT and LC-NE neurons in cardiovascular function, and the possible role of LC-NE neurons in short-and long-term volume homeostasis.     

Single unit activity of locus coeruleus neurons in the freely moving cat: I. During naturalistic behaviors and in response to simple and complex stimuli

The single unit activity of presumed noradrenergic (NE) neurons in the area of the locus coeruleus (LC) was recorded in freely moving cats. Consistent with previous reports, the activity of LC neurons was found to be state dependent: active waking greater than quiet waking greater than slow wave sleep greater than REM sleep (virtually silent). The activity of these neurons showed no relationship to movement per se. In response to simple sensory stimulation, LC units showed a short latency, short duration excitatory response. In response to a variety of non-noxious naturalistic stimuli, e.g. rats, food and a conspecific, LC unit activity did not increase above an active waking baseline. However, in response to noxious stimuli, e.g. pinches, visual threats, emesis, and forced treadmill running, LC unit activity increased above that during active waking and reached its highest levels. These data, in conjunction with those in the following report, are consistent with a general role for NE-LC neurons in the organism's adaptive response to environmental and physiological challenges.     

Noradrenergic neurons expressing substance P receptor (NK1) in the locus coeruleus complex: a double immunofluorescence study in the rat

By using a double immunofluorescence method we examined the distribution of noradrenergic neurons expressing substance P receptor (NK1) or neuromedin K receptor (NK3) in the rat brainstem. The distribution of SPR-like immunoreactive (-LI) neurons completely overlapped that of tyrosine hydroxylase (TH)-LI neurons in the locus coeruleus (A6), ventrolateral and lateral reticular formation of pons (A5 and A7). Partially overlapping distribution of SPR- and TH-LI neurons were found in certain regions of the medulla oblongata (A1-A4). Neurons showing both SPR- and TH-like immunoreactivities, however, were only found in the locus coeruleus complex (A5-A7): 100% of these TH-LI neurons displayed SPR-like immunoreactivity. Neurons showing both NKR- and TH-like immunoreactivities were not detected in the aforementioned areas of brainstem. The present study has provided morphological evidence for direct physiological modulation of noradrenergic neurons by tachykinins through SPR in locus coeruleus complex (A5-A7).     

The medial preoptic area is involved in both sexual arousal and performance in male rats: re-evaluation of neuron activity in freely moving animals

A total of 74 single unit activities was recorded from the medial preoptic-anterior hypothalamic continuum (MPOA) during free copulatory behavior of male rats. Forty-six units (62.2%) showed changed activities during at least one phase of male copulatory movements; 26 units (35.1%) increased in the firing rate during pelvic thrusting; 32 units (43.2%) increased in activity during backward jumping immediately after intromission; only one unit (1.4%) showed decreased firing rate during thrusting and backward jumping; 12 units (16.2%) increased in activity during pursuit of a female; 19 units (25.7%) were suppressed during genital grooming. Furthermore, 67 units (90.5%) showed a significant change in activity throughout a series of copulatory behavior. From the introduction of a female up to ejaculation, relatively large number of units increased in the firing rate above the value during pre-introduction adaptation period. During postejaculatory interval, however, most units decreased in activity below the level during copulation. These results strongly suggest that the MPOA is involved in both sexual arousal and performance in male rats.     

Circadian covariation of norepinephrine and serotonin in the locus coeruleus and dorsal raphe nucleus in the rat

We report robust correlations between concentrations of the neurotransmitters norepinephrine and serotonin in the locus coeruleus and the dorsal raphe nucleus of the brainstem in rats analyzed at 7 different time points over 24 h. We found similar circadian rhythmicities for both monoamines with acrophases just before the onset of the dark period. The monoamine concentrations diminished and the significant intercorrelation between norepinephrine and serotonin in the locus coeruleus disappeared during the night suggestive of a noradrenergic stimulation of dorsal raphe serotonin during the day. Timing of experiments is crucially important in studies on brain monoaminergic indices and their interrelationships.     

Participation of endogenous galanin in the suppression of baroreceptor reflex response by locus coeruleus in the rat

We evaluated the potential participation of endogenous brain galanin (GAL) in the suppression of baroreceptor reflex (BRR) response by locus coeruleus (LC), using adult male Sprague-Dawley rats anesthetized with pentobarbital sodium (40 mg/kg, i.p., with 15 mg/kg/h i.v. infusion supplements). Our physiologic and pharmacologic results demonstrated that bilateral microinjection of GAL antiserum (1:20, 20 n1) into the nucleus tractus solitarii (NTS), the terminal site for baroreceptor afferent fibers, significantly attenuated the suppressive effect of LC on the BRR response. Pretreatment with the same amount of normal rabbit serum (1:20) or heat-inactivated GAL antiserum (1:20), on the other hand, was ineffective. Microinjection of GAL (100 pmol) into the bilateral NTS also appreciably depressed the BRR response. Histochemically, retrogradely labeled neurons were distributed in the LC following microinjection of fast blue into the NTS. Immunofluorescent staining further revealed that some of these fast blue labeled LC neurons also showed positive immunoreactivity to GAL. These results suggest that a direct galaninergic projection to the NTS may participate in the suppression of BRR response by the LC.     

Vagus nerve spiking activity associated with locomotion and cortical arousal states in a freely moving rat

The vagus nerve serves as a central pathway for communication between the central and peripheral organs. Despite traditional knowledge of vagus nerve functions, detailed neurophysiological dynamics of the vagus nerve in naïve behavior remain to be understood. In this study, we developed a new method to record spiking patterns from the cervical vagus nerve while simultaneously monitoring central and peripheral organ bioelectrical signals in a freely moving rat. When the rats transiently elevated locomotor activity, the frequency of vagus nerve spikes was correspondingly increased, and this activity was retained for several seconds after the increase in running speed terminated. Spike patterns of the vagus nerve were not robustly associated with which arms the animals entered on an elevated plus maze. During sniffing behavior, vagus nerve spikes were nearly absent. During stopping, the vagus nerve spike patterns differed considerably depending on external contexts and peripheral activity states associated with cortical arousal levels. Stimulation of the vagus nerve altered rat's running speed and cortical arousal states depending on running speed at the instant of stimulation. These observations are a new step for uncovering the physiological dynamics of the vagus nerve modulating the visceral organs such as cardiovascular, respiratory, and gastrointestinal systems.     

Single unit activity of locus coeruleus neurons in the freely moving cat: II. Conditioning and pharmacologic studies

A series of experiments were carried out in which the single unit activity of presumed noradrenergic (NE) neurons in the area of the locus coeruleus (LC) was examined in freely moving cats during two conditioning paradigms and in response to pharmacologic manipulations relevant to anxiety. LC unit activity was examined during conditioned emotional response (CER) and conditioned food reward (CFR) training. During CER training, LC units showed a large increase in activity in response to a stimulus paired with a noxious air puff, whereas no increase in unit activity was seen in response to a stimulus not paired with the air puff (both comparisons relative to their appropriate control conditions). By contrast, during CFR training, a stimulus paired with a rewarding food delivery did not elicit a significant increase in LC unit activity relative to its appropriate control condition. Therefore, NE units in the LC greatly increase their activity in response to a stimulus that predicts the occurrence of a noxious event, but not in response to a stimulus that predicts the occurrence of a rewarding event. Administration of the anxiolytic drug diazepam (0.25 and 2.0 mg/kg, i.p.) had no significant effect on the spontaneous activity of LC neurons, but significantly reduced the excitatory response of LC neurons to simple stimuli. Administration of the anxiogenic drug yohimbine (2.0 mg/kg, i.p.) significantly increased the spontaneous activity of LC neurons. These results support the hypothesis that LC neurons play a role in aversive but not appetitive conditioning, and are consistent with the hypothesis that these neurons may play a role in anxiety. These data are discussed within the broader context of LC neurons mediating the organism's adaptive response to environmental or physiological challenges.     

Afferent projections to the self-stimulation regions of the dorsal pons, including the locus coeruleus, in the rat as demonstrated by the horseradish peroxidase technique

Afferent projections to the dorsal pons of the rat have been studied using the horseradish peroxidase (HRP) technique. HRP injections were made in each of the following regions: the vicinity of the locus coeruleus (LC); the periventncular gray, medial to the LC; the medial parabrachial region, lateral to the LC; the ventral cerebellum, dorsal to the LC; and the pontine reticular formation, ventral to the LC. Because intracranial self-stimulation (ICSS) has been obtained in these regions, the afferents have been discussed in terms of their possible contributions to that behavior. Previous ICSS studies of the dorsal pons have focussed on the LC as playing a central role. Presently identified inputs to the LC include: the dorsal raphe nucleus: the ventrolateral periaqueductal gray: the pontine reticular formation: the areas that contain the pontine and medullary noradrenergic and adrenergic cell groups: the lateral hypothalamic area: the contralateral LC: the deep cerebellar nuclei: the ventrolateral and parafascicular thalamic nuclei: and the parabrachial regions of the pons and midbrain.     

Depression as a spreading adjustment disorder of monoaminergic neurons: a case for primary implication of the locus coeruleus

A model for the pathophysiology of depression is discussed in the context of other existing theories. The classic monoamine theory of depression suggests that a deficit in monoamine neurotransmitters in the synaptic cleft is the primary cause of depression. More recent elaborations of the classic theory also implicitly include this postulate, other theories of depression frequently prefer to depart from the monoamine-based model altogether. We suggest that the primary defect emerges in the regulation of firing rates in brainstem monoaminergic neurons, which brings about a decrease in the tonic release of neurotransmitters in their projection areas, an increase in postsynaptic sensitivity, and concomitantly, exaggerated responses to acute increases in the presynaptic firing rate and transmitter release. It is proposed that the initial defect involves, in particular, the noradrenergic innervation from the locus coeruleus (LC). Dysregulation of the LC projection activities may lead in turn to dysregulation of serotonergic and dopaminergic neurotransmission. Failure of the LC function could explain the basic impairments in the processing of novel information, intensive processing of irrational beliefs, and anxiety. Concomitant impairments in the serotonergic neurotransmission may contribute to the mood changes and reduction in the mesotelencephalic dopaminergic activity to loss of motivation, and anhedonia. Dysregulation of CRF and other neuropeptides such as neuropeptide Y, galanin and substance P may reinforce the LC dysfunction and thus further weaken the adaptivity to stressful stimuli.     

Functional role of alpha1-adrenoceptors in the locus coeruleus: a microdialysis study

The present study elucidates the role of alpha(1)-adrenoreceptors in the locus coeruleus (LC) using a dual-probe microdialysis in conscious rats. One probe sampled noradrenaline in the LC, whereas the second probe sampled noradrenaline in a main projection area, the prefrontal cortex (PFC). To investigate a possible tonic activation of LC neurons by alpha(1)-adrenoceptor, the alpha(1)-antagonist prazosin (10 microM) was infused into the LC. Extracellular noradrenaline in the LC decreased to about 50% of basal levels but no change of noradrenaline release was detected in the ipsilateral PFC. Next, the interaction between alpha(1)- and alpha(2)-adrenoceptors was investigated. Local administration of the alpha(2)-adrenoceptor antagonist idazoxan (100 microM) into the LC increased the noradrenaline release in the LC to about 400%, whereas noradrenaline release in the PFC rose to 150% of basal levels. A similar effect was seen when the specific alpha(2A)-adrenoceptor antagonist BRL 44408 (10 microM) was infused: extracellular noradrenaline in the LC and PFC increased to about 400 and 120% of the basal levels, respectively. When infusions of idazoxan (100 microM) or BRL 44408 (10 microM) into the LC were combined with prazosin (10 microM), the excitatory effects of the alpha(2)-adrenoceptor antagonists on the release of noradrenaline were strongly suppressed in the LC as well as in the ipsilateral PFC. It is concluded that alpha(1)-adrenoreceptors are involved in the regulation of LC activity. Apparently, alpha(1)- and alpha(2)-adrenoceptors have opposite roles in their function as autoreceptors on LC cells.     

Monoamine oxidase activity in noradrenaline neurons of the locus coeruleus of the rat. A double-labeling histochemical study

The aim of the present study is to examine whether noradrenergic neurons of the locus coeruleus (LC) of the rat contain monoamine oxidase (MAO) activity. Sections were processed initially for MAO enzyme histochemistry using tyramine as a substrate, followed by fluorescence immunohistochemistry for tyrosine hydroxylase (TH). In the LC, virtually all TH-immunoreactive neurons (i.e., noradrenergic neurons) were also positive for MAO. No MAO activity was found in any TH-negative neurons. Neurons in the LC have previously been shown to form dopamine during noradrenaline biosynthesis and to produce serotonin from exogenously administered -5-hydroxytryptophan. Moreover, dopamine- and serotonin-degrading MAO activity has also been found in LC neurons. Therefore, our results indicate that MAO activity is localized within noradrenergic neurons in the LC and is likely involved in the degradation of dopamine that is endogenously synthesized, and also in the elimination of serotonin that is produced from exogenous precursors.     

Spinal projections from the nucleus locus coeruleus and nucleus subcoeruleus in the cat and monkey as demonstrated by the retrograde transport of horseradish peroxidase

The rostral pons of the cat and rhesus monkey were examined for the presence of labeled cells following injections of horseradish peroxidase (HRP) into the lumbar spinal cord. Labeled cells were found in the ipsilateral dorsolateral pontine tegmentum and in the contralateral ventrolateral pontine reticular formation. In both the cat and monkey, labeled cells were located in the nucleus locus coeruleus, nucleus subcoeruleus, in or near the Kölliker-Fuse nucleus, and in the ventral part of the lateral parabrachial nucleus. There is a striking similarity between the distribution of HRP-labeled cells in the dorsolateral pontine tegmentum of the cat and monkey and that of catecholamine-containing cells observed in this area in previous studies.     

Selective prefrontal cortical projections to the region of the locus coeruleus and raphe nuclei in the rhesus monkey

Descending cortical projections to the region of the noradrenergic locus coeruleus (LC) and the serotonergic dorsal and central superior raphe nuclei were analyzed in the rhesus monkey using anterograde labeling techniques. HRP pellets or tritiated leucine were injected into one of 7 cortical areas: the dorsolateral prefrontal cortex, the dorsomedial prefrontal cortex, the orbital prefrontal cortex, the parietal association cortex, somatosensory cortex, the anterior portion of the inferior temporal gyrus, and the posterior portion of the inferior temporal gyrus. Anterogradely labeled fibers were found in and adjacent to the LC and raphe nuclei only following the dorsolateral and dorsomedial prefrontal cortical injections. Terminal labeling was densest at rostral levels of the LC, particularly in the area directly medial to the nucleus. Labeled fibers could not be followed beyond caudal levels of the LC. The projections to the contralateral LC and raphe nuclei were similar to, but less dense than that to the ipsilateral region. Injections into cortical areas other than the dorsal prefrontal cortex resulted in anterograde labeling of the pontine nuclei or pyramids, but not the LC/raphe region. These data, in conjunction with studies in the rat, suggest that the dorsal prefrontal cortex may be the only cortical area to have direct influence on the LC and raphe nuclei and secondary influence on the monoaminergic innervation of large areas of cerebral cortex.