Effects of coagulation of thelocus coeruleus and pontine raphe nuclei on the background activity recorded from the rat cerebellar fastigial nucleus neurons

Simultaneous or separate coagulation of thelocus coeruleus (LC) and the pontine raphe nucleus (PRN) results in a significant increase of irregular-type background activity in the cerebellar fastigial nucleus neurons. There are also considerable changes in the dynamics of impulse sequences, in particular, the number of neurons with random interpulse intervals markedly increases. Destruction of theLC and/or PRN is followed by a marked drop in the mean frequency of discharges in the neurons of the fastigial nucleus.Neirofiziologiya/Neurophysiology, Vol. 26, No. 6, pp. 437–442, November–December, 1994.     

Serotonin augments the cationic current Ih in central neurons

Serotonin (5-HT) induced a slow depolarization when superfused onto neurons of the rat brainstem nucleus prepositus hypoglossi (PH) in vitro. The depolarization was associated with a decrease in cell input resistance. In voltage clamp, 5-HT caused an inward current that activated at approximately -50 mV and was present only at potentials negative to this. With hyperpolarizing voltage-clamp steps, PH neurons exhibited a slow inward current relaxation. The properties of this conductance were consistent with the cationic, nonselective current, Ih. Bath-applied 5-HT augmented Ih. Extracellular CsCl blocked both Ih and the inward current produced by 5-HT. In addition, forskolin, isobutylmethylxanthine, and 8-bromo-cAMP mimicked the inward current seen with 5-HT. The 5-HT1 agonist 5-carboxamidotryptamine produced a similar inward current. We conclude that 5-HT excites PH neurons by augmenting Ih, probably through receptor-mediated stimulation of adenylate cyclase. As Ih is found in many types of neurons, this mechanism may be a common mode of regulating cell excitability.     

Afferent connections of the cat lateral vestibular nucleus

Location within the brain of HP-labeled neurons (origins of projections to the lateral vestibular nucleus) was investigated by iontophoretic injection of this enzyme. Bilateral projections to the following midbrain structures were revealed: the field of Forel, interstitial nuclei of Cajal, oculomotor nerve nuclei, and the red nucleus — to all parts of the lateral vestibular nucleus. Bilateral projections were also shown from more caudally located structures, viz. the superior, medial and inferior (descending) vestibular nuclei, Y groups of the vestibular nuclear complex, facial nucleus and hypoglossi, nucleus prepositus nervi hypoglossi and caudal nuclei of the trigeminal tract; ipsilateral projections from crus IIa of lobulus ansiformus of the cerebellar hemisphere; contralateral projections from the bulbar lateral reticular nucleus and Deiter's nucleus. A tonic organization pattern of afferent inputs from a number of brainstem formations to the dorsal and ventral lateral vestibular nucleus is revealed and trajectories of HP-labeled fiber systems projecting to Deiter's nucleus described.L. A. Orbeli Institute of Physiology, Academy of Sciences of the Armenian SSR, Erevan. Translated from Neirofiziologiya, Vol. 20, No. 4, pp. 494–503, July–August, 1988.     

Efferent projections of the area postrema demonstrated by autoradiography

The efferents connections of the area postrema (AP) have been studied autoradiographically following iontophoretic injections of 3H-glycine or 3H-leucine into the area postrema. Precise control of the diffusion of the labelled amino acids injected iontophorectically into the AP was made using the technique of sectioning with a cryostat. AP projects to a great number of structures. Projections to nucleus tractus solitarius (NFS), dorsal vagal nucleus, nucleus intercalatus, nucleus praepositus hypoglossi, nucleus hypoglossal, the mesencephalic nucleus of V nerve, locus coeruleus and superior and inferior colliculi are shown bilaterally. The density of the efferents was greatest to the NFS and the LC. Corelations are suggested with functional mechanisms of cardiovascular regulation.     

Benzodiazepines attenuate single unit activity in the locus coeruleus

Several classes of anxiolytic compounds have the common effect of decreasing the firing of noradrenergic neurons or attenuating the post- synaptic effects of noradrenergic activity. In order to determine whether the benzodiazepines, the most widely used anxiolytics, also decrease noradrenergic activity, the effect of acute intravenous injections of diazepam (0.1–2.0 mg/kg) and chlordiazepoxide (0.5–4.0 mg/kg) were administered to anesthetized rats while spontaneous activity of single neurons in the principal noradrenergic nucleus, the locus coeruleus, was recorded. Diazepam and chlordiazepoxide decreased spontaneous single unit activity in the locus coeruleus at relatively low doses. This net effect on noradrenergic systems is consistent with the actions of several classes of nonbenzodiazepine anxiolytics, and with the involvement of noradrenergic systems in the neural mechanisms of anxiety.     

Afferent connections of cat facial nucleus; a study using retrograde axonal transport of horseradish peroxidase

Neuronal populations in the brainstem and spinal cord — the sources of fiber pathways to the facial nucleus — were investigated in adult cats by microiontophoretically injecting horseradish peroxidase into restricted areas of the facial nucleus. Projections were identified from thenucleus nervi hypoglossi, nucleus praepositus hypoglossi, nucleus raphe pallidus, nucleus intercalatus, medial nucleus of the solitary tract, dorsal motor nucleus of the vagus, neurons of genu of the facial nerve, ipsilateral red nucleus, and reticular formation of the midbrain to the facial nucleus. Projections from a number of other brain structures to the facial nucleus also received confirmation. A topographic map was drawn up, showing how brainstem and spinal cord afferents are distributed in the facial nucleus.L. A. Orbeli Institute of Physiology, Academy of Sciences of the Armenian SSR, Erevan. Translated from Neirofiziologiya, Vol. 18, No. 1, pp. 35–45, January–February, 1986.     

Vestibular integrator neurons have quadratic functions due to voltage dependent conductances

The nonlinear properties of the dendrites of the prepositus hypoglossi nucleus (PHN) neurons are essential for the operation of the vestibular neural integrator that converts a head velocity signal to one that controls eye position. A novel system of frequency probing, namely quadratic sinusoidal analysis (QSA), was used to decode the intrinsic nonlinear behavior of these neurons under voltage clamp conditions. Voltage clamp currents were measured at harmonic and interactive frequencies using specific nonoverlapping stimulation frequencies. Eigenanalysis of the QSA matrix reduces it to a remarkably compact processing unit, composed of just one or two dominant components (eigenvalues). The QSA matrix of rat PHN neurons provides signatures of the voltage dependent conductances for their particular dendritic and somatic distributions. An important part of the nonlinear response is due to the persistent sodium conductance (g_NaP), which is likely to be essential for sustained effects needed for a neural integrator. It was found that responses in the range of 10 mV peak to peak could be well described by quadratic nonlinearities suggesting that effects of higher degree nonlinearities would add only marginal improvement. Therefore, the quadratic response is likely to sufficiently capture most of the nonlinear behavior of neuronal systems except for extremely large synaptic inputs. Thus, neurons have two distinct linear and quadratic functions, which shows that piecewise linear?+?quadratic analysis is much more complete than just piecewise linear analysis; in addition quadratic analysis can be done at a single holding potential. Furthermore, the nonlinear neuronal responses contain more frequencies over a wider frequency band than the input signal. As a consequence, they convert limited amplitude and bandwidth input signals to wider bandwidth and more complex output responses. Finally, simulations at subthreshold membrane potentials with realistic PHN neuron models suggest that the quadratic functions are fundamentally dominated by active dendritic structures and persistent sodium conductances.     

Role of the lateral paragigantocellular nucleus in the network of paradoxical (REM) sleep: an electrophysiological and anatomical study in the rat

The lateral paragigantocellular nucleus (LPGi) is located in the ventrolateral medulla and is known as a sympathoexcitatory area involved in the control of blood pressure. In recent experiments, we showed that the LPGi contains a large number of neurons activated during PS hypersomnia following a selective deprivation. Among these neurons, more than two-thirds are GABAergic and more than one fourth send efferent fibers to the wake-active locus coeruleus nucleus. To get more insight into the role of the LPGi in PS regulation, we combined an electrophysiological and anatomical approach in the rat, using extracellular recordings in the head-restrained model and injections of tracers followed by the immunohistochemical detection of Fos in control, PS-deprived and PS-recovery animals. With the head-restrained preparation, we showed that the LPGi contains neurons specifically active during PS (PS-On neurons), neurons inactive during PS (PS-Off neurons) and neurons indifferent to the sleep-waking cycle. After injection of CTb in the facial nucleus, the neurons of which are hyperpolarized during PS, the largest population of Fos/CTb neurons visualized in the medulla in the PS-recovery condition was observed in the LPGi. After injection of CTb in the LPGi itself and PS-recovery, the nucleus containing the highest number of Fos/CTb neurons, moreover bilaterally, was the sublaterodorsal nucleus (SLD). The SLD is known as the pontine executive PS area and triggers PS through glutamatergic neurons. We propose that, during PS, the LPGi is strongly excited by the SLD and hyperpolarizes the motoneurons of the facial nucleus in addition to local and locus coeruleus PS-Off neurons, and by this means contributes to PS genesis.     

Identification of a Group of GABAergic Neurons in the Dorsomedial Area of the Locus Coeruleus

The locus coeruleus (LC)-norepinephrine (NE) system in the brainstem plays a critical role in a variety of behaviors is an important target of pharmacological intervention to several neurological disorders. Although GABA is the major inhibitory neurotransmitter of LC neurons, the modulation of LC neuronal firing activity by local GABAergic interneurons remains poorly understood with respect to their precise location, intrinsic membrane properties and synaptic modulation. Here, we took an optogenetic approach to address these questions. Channelrhodopsin (ChR2) in a tandem with the yellow fluorescent protein (YFP) was expressed in GABAergic neurons under the control of glutamic acid decarboxylase 2 (GAD2) promoter. Immediately dorsomedial to the LC nucleus, a group of GABAergic neurons was observed. They had small soma and were densely packed in a small area, which we named the dorsomedial LC or dmLC nucleus. These GABAergic neurons showed fast firing activity, strong inward rectification and spike frequency adaptation. Lateral inhibition among these GABAergic neurons was observed. Optostimulation of the dmLC area drastically inhibited LC neuronal firing frequency, expanded the spike intervals, and reset their pacemaking activity. Analysis of the light evoked inhibitory postsynaptic currents (IPSCs) indicated that they were monosynaptic. Such light evoked IPSCs were not seen in slices where this group of GABAergic neurons was absent. Thus, an isolated group of GABAergic neurons is demonstrated in the LC area, whose location, somatic morphology and intrinsic membrane properties are clearly distinguishable from adjacent LC neurons. They interact with each and may inhibit LC neurons as well as a part of local neuronal circuitry in the LC.     

Neuroanatomical localization of substance p: implications for central cardiovascular control

The presence of substance P (SP) neurons in pathways known to be involved in the central control of the cardiovascular system has been studied with neuroanatomical and neurochemical techniques. SP-immunoreactive (SP-I) neurons are found in afferent baro- and chemoreceptor pathways which transmit information from peripheral receptors to the nucleus tractus solitarius. In addition, SP-I neurons located in the nucleus interfascicularis hypoglossi of the ventral medulla innervate the intermediolateral cell column, the site of origin of preganglionic sympthetic nerves. The role of these SP-I neurons in cardiovascular control remains to be determined.     

蓝斑核和中缝背核参与电剌激弓状核引起的大鼠...

The effect of electrical stimulation of hypothalamic arcuate nucleus (ARC) on intragastric pressure (IGP) was observed on 80 Wistar rats anaesthetized with urethan. The main results are as follows: (1) Electrical stimulation of ARC could cause an obvious decrease of IGP. (2) The reduction of IGP induced by electrical stimulation of ARC was not affected by intracerebroventricular injection of naloxone. (3) After lesioning of locus coeruleus or dorsal raphe, the effect of ARC stimulation was depressed. The results suggest that the locus coeruleus and dorsal raphe nucleus may be involved in the reduction of IGP induced by ARC stimulation, but without the involvement of beta-endorphinergic neurons.     

Afferent pathway from locus coeruleus to the nucleus solitarius

After the destruction of the nucleus tractas solitarii, just caudally to the writing pen by means of a stereotaxic instrument, the system of afferent fibres to the nucleus in question was investigated by the methods of Nauta and Fink--Heimer. The fibre terminals were revealed near locus coeruleus. Investigation of the locus coeruleus by Golgi method demonstrated that it usually has neurons of reticular type and transitional ones which resemble by their form the neurons specific for sensory formations. It is possible to conclude that locus coeruleus posesses connections of visceral origin which may play a part in the afferent influence of locus coeruleus on the brain cortex.     

Nonuniformity in the linear network model of the oculomotor integrator produces approximately fractional-order dynamics and more realistic neuron behavior

The oculomotor integrator is a network that is composed of neurons in the medial vestibular nuclei and nuclei prepositus hypoglossi in the brainstem. Those neurons act approximately as fractional integrators of various orders, converting eye velocity commands into signals that are intermediate between velocity and position. The oculomotor integrator has been modeled as a network of linear neural elements, the time constants of which are lengthened by positive feedback through reciprocal inhibition. In this model, in which each neuron reciprocally inhibits its neighbors with the same Gaussian profile, all model neurons behave as identical, first-order, low-pass filters with dynamics that do not match the variable, approximately fractional-order dynamics of the neurons that compose the actual oculomotor integrator. Fractional-order integrators can be approximated by weighted sums of first-order, low-pass filters with diverse, broadly distributed time constants. Dynamic systems analysis reveals that the model integrator indeed has many broadly distributed time constants. However, only one time constant is expressed in the model due to the uniformity of its network connections. If the model network is made nonuniform by removing the reciprocal connections to and from a small number of neurons, then many more time constants are expressed. The dynamics of the neurons in the nonuniform network model are variable, approximately fractional-order, and resemble those of the neurons that compose the actual oculomotor integrator. Completely removing the connections to and from a neuron is equivalent to eliminating it, an operation done previously to demonstrate the robustness of the integrator network model. Ironically, the resulting nonuniform network model, previously supposed to represent a pathological integrator, may in fact represent a healthy integrator containing neurons with realistically variable, approximately fractional-order dynamics. Received: 8 August 1997 / Accepted in revised form: 18 June 1998     

Alpha-neo-endorphin-like immunoreactivity in the cat brain stem

This paper examines the distribution of fibers and cell bodies containing alpha-neo-endorphin in the cat brain stem by using an indirect immunoperoxidase technique. A high or moderate density of immunoreactive cell bodies was found in the superior central nucleus, nucleus incertus, dorsal tegmental nucleus, nucleus of the trapezoid body, and in the laminar spinal trigeminal nucleus, whereas a low density of such perikarya was observed in the inferior colliculus, nucleus praepositus hypoglossi, dorsal nucleus of the raphe, nucleus of the brachium of the inferior colliculus, and in the nucleus of the solitary tract. The highest density of immunoreactive fibers was found in the substantia nigra, dorsal motor nucleus of the vagus, nucleus coeruleus, lateral tegmental field, marginal nucleus of the brachium conjunctivum, and in the inferior and medial vestibular nuclei. These results indicate that alpha-neo-endorphin is widely distributed in the cat brain stem and suggest that the peptide could play an important role in several physiological functions, e.g., those involved in respiratory, cardiovascular, auditory, and motor mechanisms.     

Localization of cholinergic neurons in the cat lower brain stem

The localization of cholinergic neurons in the cat lower brain stem was determined immunocytochemically with a monoclonal antibody against choline acetyltransferase (ChAT), the acetylcholine synthesizing enzyme. ChAT-positive neurons were observed in four major cell groups: cranial nerve motor and special visceromotor neurons: parasympathetic preganglionic visceromotor neurons; neurons located in the ponto-mesencephalic tegmentum including area X (or pedunculopontine tegmental nucleus), nucleus laterodorsalis tegmenti (Ldt) of Castaldi, and peri-locus coeruleus alpha (peri-alpha); and neurons located in nucleus reticularis magnocellularis (Mc) and adjacent nucleus reticularis gigantocellularis (Gc) of the medulla.     

Altered serotonin and dopamine metabolism in the CNS of serotonin 5-HT(1A) or 5-HT(1B) receptor knockout mice

Measurements of serotonin (5-HT), dopamine (DA), and noradrenaline, and of 5-HT and DA metabolites, were obtained by HPLC from 16 brain regions and the spinal cord of 5-HT(1A) or 5-HT(1B) knockout and wild-type mice of the 129/Sv strain. In 5-HT(1A) knockouts, 5-HT concentrations were unchanged throughout, but levels of 5-HT metabolites were higher than those of the wild type in dorsal/medial raphe nuclei, olfactory bulb, substantia nigra, and locus coeruleus. This was taken as an indication of increased 5-HT turnover, reflecting an augmented basal activity of midbrain raphe neurons and consequent increase in their somatodendritic and axon terminal release of 5-HT. It provided a likely explanation for the increased anxious-like behavior observed in 5-HT(1A) knockout mice. Concomitant increases in DA content and/or DA turnover were interpreted as the result of a disinhibition of DA, whereas increases in noradrenaline concentration in some territories of projection of the locus coeruleus could reflect a diminished activity of its neurons. In 5-HT(1B) knockouts, 5-HT concentrations were lower than those of the wild type in nucleus accumbens, locus coeruleus, spinal cord, and probably also several other territories of 5-HT innervation. A decrease in DA, associated with increased DA turnover, was measured in nucleus accumbens. These changes in 5-HT and DA metabolism were consistent with the increased aggressiveness and the supersensitivity to cocaine reported in 5-HT(1B) knockout mice. Thus, markedly different alterations in CNS monoamine metabolism may contribute to the opposite behavioral phenotypes of these two knockouts.     

Chronic Alterations in Monoaminergic Cells in the Locus Coeruleus in Orexin Neuron-Ablated Narcoleptic Mice

Narcolepsy patients often suffer from insomnia in addition to excessive daytime sleepiness. Narcoleptic animals also show behavioral instability characterized by frequent transitions between all vigilance states, exhibiting very short bouts of NREM sleep as well as wakefulness. The instability of wakefulness states in narcolepsy is thought to be due to deficiency of orexins, neuropeptides produced in the lateral hypothalamic neurons, which play a highly important role in maintaining wakefulness. However, the mechanism responsible for sleep instability in this disorder remains to be elucidated. Because firing of orexin neurons ceases during sleep in healthy animals, deficiency of orexins does not explain the abnormality of sleep. We hypothesized that chronic compensatory changes in the neurophysiologica activity of the locus coeruleus (LC) and dorsal raphe (DR) nucleus in response to the progressive loss of endogenous orexin tone underlie the pathological regulation of sleep/wake states. To evaluate this hypothesis, we examined firing patterns of serotonergic (5-HT) neurons and noradrenergic (NA) neurons in the brain stem, two important neuronal populations in the regulation of sleep/wakefulness states. We recorded single-unit activities of 5-HT neurons and NA neurons in the DR nucleus and LC of orexin neuron-ablated narcoleptic mice. We found that while the firing pattern of 5-HT neurons in narcoleptic mice was similar to that in wildtype mice, that of NA neurons was significantly different from that in wildtype mice. In narcoleptic mice, NA neurons showed a higher firing frequency during both wakefulness and NREM sleep as compared with wildtype mice. In vitro patch-clamp study of NA neurons of narcoleptic mice suggested a functional decrease of GABAergic input to these neurons. These alterations might play roles in the sleep abnormality in narcolepsy.     

Observations on the projection from the perihypoglossal nuclei onto the cerebellum in the macaque monkey

The occurrence and distribution of retrogradely labeled cells in the perihypoglossal nuclei of the monkey were mapped after injections of horseradish peroxidase in various cerebellar cortical regions. In general the findings are in accord with those made in the cat. The flocculus receives a heavy bilateral projection from the nucleus prepositus, particularly from its caudoventral part, and from the nucleus of Roller. There is an apparently scanty projection from the nucleus intercalatus. The uvula receives a rather similar projection, but in the prepositus the cells projecting to the uvula are on the whole situated more dorsally and rostrally than those supplying the flocculus. The projection to lobules VII-VIII is distinct. More scanty projections have been found to the paramedian lobule and the anterior lobe. The different but partially overlapping sites of origin in the prepositus of fibers to the flocculus and uvula indicate the presence of a topical pattern within the perihypoglosso-cerebellar projection, as in the cat (34). In the monkey the two regions of origin appear to coincide with two particular cell collections in the prepositus (12). Both small and middle sized cells project to the cerebellum, as they do in the cat (9, 48). The nucleus supragenualis nervi facialis in the macaque is morphologically different from the corresponding nucleus in most other mammalian species (12), but it contains labeled cells after injections in the flocculus, uvula and other cerebellar regions. A considerable number of cells in the abducent nucleus are labeled after injections in the flocculus and the posterior vermis.     

6-Hydroxydopamine treatment enhances excitation of cultured cerebellar neurons by glutamate

Explant cultures containing identifiable cerebellar cortical neurons and locus coeruleus neurons were treated with 500 microM 6-hydroxydopamine. At this concentration, locus coeruleus neurons were usually degenerated after 48 h, while the cerebellar cortical neurons had a normal appearance. Extracellular recording and iontophoresis of noradrenaline and glutamate were used to test for changes in electrical activity or neurotransmitter responsiveness of the cerebellar neurons. At 4-5 days following the toxin exposure, spontaneous spiking activity appeared similar to that in control cultures. Mean iontophoretic currents required to give noradrenaline-induced depressions of activity were somewhat lower for the toxin-treated cultures than for controls but not significantly so. The mean currents for glutamate excitations, however, were markedly lower in the treated cultures. Noradrenaline potentiations of glutamate responses were observed in both treated and control cultures. The greatly increased sensitivity of cerebellar neurons to glutamate does not seem related to degeneration of granule cells in the treated cultures but might be explained by disruption of astrocytic uptake mechanisms for glutamate.     

The spontaneous neuronal activity of the hippocampus during the modulation of the theta rhythm by cholinergic substances]

Background activity of the hippocampal neurons, extracellularly recorded in waking chronic rabbits, was analysed in control state and after systemic injection of physostigmine and scopolamine. Similar analysis was done in the hippocampus chronically deprived of ascending brain stem afferents. Cholinergic drugs controlled the number of hippocampal neurons with theta-modulation and the degree of its stability but not the frequency. Activation of cholinergic theta-rhythm resulted also in regularization of activity with suppression of delta-modulation and complex spike discharges; its blockade was accompanied by the opposite changes. Both drugs shifted the level of background activity in the majority of neurons, but the overall mean frequency did not vary between the states. Regression analysis demonstrated significant negative correlations with dominating decrease in the level of activity in high-frequency neurons ( > 25 sp/s) and its increase in low-frequency ones ( < 25 sp/s) after injection of both drugs. Stability of the overall mean frequency and uniformity of its shifts presumably indicate that the frequency, unlike the pattern of the background activity, is not directly controlled by the cholinergic septal input.