Nucleus Reticularis Magnocellularis Research Articles

Evaluation of cellular activity in response to sleep deprivation by a comprehensive analysis of the whole mouse brain.

Sleep deprivation (SD) causes several adverse functional outcomes, and understanding the associated processes can improve quality of life. Although the effects of SD on neuronal activity in several brain regions have been identified, a comprehensive evaluation of the whole brain is still lacking. Hence, we performed SD using two different methods, gentle handling and a dedicated chamber, in targeted recombination in active populations 2 (TRAP2) mice crossed with Rosa-ZsGreen reporter mice and visualized cellular activity in the whole brain. Using the semi-automated post-imaging analysis tool Slice Histology Alignment, Registration, and Cell Quantification (SHARCQ), the number of activated cells was quantified. From the analysis of 14 brain regions, cellular activity was significantly increased in the olfactory areas and decreased in the medulla by the two SD methods. From the analysis of the further subdivided 348 regions, cellular activity was significantly increased in the vascular organ of the lamina terminalis, lateral hypothalamic area, parabigeminal nucleus, ventral tegmental area, and magnocellular reticular nucleus, and decreased in the anterior part of the basolateral amygdalar nucleus, nucleus accumbens, septohippocampal nucleus, reticular nucleus of the thalamus, preoptic part of the periventricular hypothalamic nucleus, ventromedial preoptic nucleus, rostral linear nucleus raphe, facial motor nucleus, vestibular nuclei, and some fiber tracts (oculomotor nerve, genu of corpus callosum, and rubrospinal tract) by the two SD methods. Two subdivided regions of the striatum (caudoputamen and other striatum), epithalamus, vascular organ of the lamina terminalis, anteroventral preoptic nucleus, superior colliculus optic layer, medial terminal nucleus of the accessory optic tract, pontine gray, and fiber tracts (medial lemniscus, columns of the fornix, brachium of the inferior colliculus, and mammillary peduncle) were differentially affected by the two SD methods. Most brain regions detected from these analyses have been reported to be involved in regulating sleep/wake regulatory circuits. Moreover, the results from the connectivity analysis indicated that the connectivity of cellular activity among brain regions was altered by SD. Together, such a comprehensive analysis of the whole brain is useful for understanding the mechanisms by which SD and/or sleep disruption affects brain function.

Sex and estrogen affect the distribution of urocortin-1 immunoreactivity in brainstem autonomic nuclei of the rat.

Urocortin-1 (UCN-1), a neuropeptide closely related to the hypothalamic hormone corticotropin-releasing factor, has been associated with stress, feeding behaviors, cardiovascular control, and to exhibit functional gender differences. This study was done to investigate whether estrogen (E; 17β-estradiol) treatment (9 weeks) altered UCN-1 immunoreactivity in brainstem autonomic nuclei in female Wistar rats. Experiments were done in age matched adult males (controls), females (intact), and ovariectomized (OVX) only and OVX+E (30pg/ml plasma) treated females. All animals received intracerebroventricular injections of colchicine and were then perfused transcardially with Zamboni's fixative. Coronal brainstem sections (40μm) were cut and processed immunohistochemically for UCN-1. In males, moderate UCN-1 fiber labeling was found in the nucleus of the solitary tract (NTS) and throughout the rostral ventral lateral medulla (RVLM). Additionally, a few UCN-1 immunoreactive neurons were observed in hypoglossal nucleus (XII), facial nucleus (FN) and nucleus ambiguus (Amb). In intact females and OVX+E females, fewer UCN-1 labeled fibers were found within NTS compared to males. In contrast, the RVLM was more densely innervated in the female cases. Furthermore, in both intact and OVX+E females UCN-1 labeled neurons were found not only within Amb, FN and XII, but also within NTS, RVLM and nucleus raphé pallidus (RP). In OVX only animals, moderate to dense UCN-1 fiber labeling was observed in the NTS complex and throughout RVLM compared to males and the other female groups. However, in contrast to all other groups, UCN-1 labeled neurons were found in greater number within Amb, FN, NTS, dorsal motor nucleus of the vagus, XII, RVLM, magnocellular reticular nucleus and RP. These data not only suggest that sex differences exist in the distribution of UCN-1 within brainstem autonomic areas, but that circulating level of E may play an important role with regards to the function of these UCN-1 neurons during stress responses.

Brainstem neurons responsible for postural, masseter or pharyngeal muscle atonia during paradoxical sleep in freely-moving cats.

In this mini review, we summarize our findings regarding the brainstem neurons responsible for the postural, masseter, or pharyngeal muscle atonia observed during paradoxical sleep (PS) in freely moving cats. Both the pons and medulla contain neurons showing tonic activation selective to PS and atonia, referred to as PS/atonia-on-neurons. The PS/atonia-on neurons, characterized by their most slow conducting property and located in the peri-locus coeruleus alpha (peri-LCa) and adjacent LCa of the mediodorsal pontine tegmentum, play a critical executive role in the somatic and orofacial muscle atonia observed during PS. Slow conducting medullary PS/atonia-on neurons located in the nuclei reticularis magnocellularis (Mc) and parvocellularis (Pc) may play a critical executive role in the generation of, respectively, antigravity or orofacial muscle atonia during PS. In addition, either tonic or phasic cessation of activity of medullary serotonergic neurons may play an important role in the atonia of genioglossus muscles during PS via a mechanism of disfacilitation.

Collateral axonal projections from rostral ventromedial medullary nitric oxide synthase containing neurons to brainstem autonomic sites

The magnocellular reticular nucleus and adjacent lateral paragigantocellular nucleus have been shown to contain a large population of nitric oxide synthase (NOS) immunoreactive neurons. However, little is known about the projections of these neurons within the central nervous system. Retrograde tract-tracing techniques combined with immunohistochemistry were used in this study to investigate whether NOS neurons in this rostral ventromedial medullary (RVMM) region send collateral axonal projections to autonomic sites in the nucleus of the solitary tract (NTS) and in the nucleus ambiguus (Amb). Fluorogold and/or rhodamine labeled latex microspheres were microinjected into the NTS and Amb at sites that elicited bardycardia and/or depressor responses ( l-glutamate; 0.25 M; 10 nl). After a survival period of 10–14 days, the rats were sacrificed and tissue sections of the brainstem were processed immunohistochemically for the identification of NOS containing neuronal perikarya. After unilateral injection of the tract-tracers into the NTS and Amb, retrogradely labeled neurons were observed bilaterally throughout the RVMM region. Of the number of RVMM neurons retrogradely labeled from the NTS (684 ± 143), 9% were found to be immunoreactive to NOS. Similarly, of those RVMM neurons retrogradely labeled from the Amb (963 ± 207), 7% also contained NOS immunoreactivity. Neurons with collateral axonal projections to NTS and Amb (14% and 10%, respectively) were observed predominantly within a region of RVMM that extended co-extensively with approximately the rostrocaudal extent of the facial nucleus. Of these double labeled neurons, 36.4 ± 20 (39%) were also found to be immunoreactive to NOS. These data indicate that the RVMM contains at least three population of NOS neurons that send axons to innervate functionally similar cardiovascular responsive sites in the NTS and Amb. Although the function of these NOS containing medullary pathways in cardiovascular control is not known, it is likely that those with collateral axonal projections represent the anatomical substrate by which the RVMM may simultaneously coordinate cardiovascular responses during physiological changes associated with respiration and/or motor movements.

THE EFFECT OF GENDER ON THE DISTRIBUTION OF UROCORTIN NEURONS IN THE BRAINSTEM

Intraventricular (icv) administration of urocortin (ucn), a neuropeptide closely related to the hypothalamic hormone corticotropin-releasing factor, has been shown to elicit transient sympathoexcitatory responses in the male rat. However, the central sites at which ucn exerts these effects and whether gender differences exist in the location and distribution of ucn neurons are not known. In this study, experiments were done in male, and in female ovariectomized (OVX) only and OVX 17β-estradiol (30 pg/ml plasma; OVX+E) treated Wistar rats to investigate the distribution of neurons in the brainstem containing ucn-like immunoreactivity. Six to 9 weeks after OVX or OVX+E, animals were given an icv injection of colchicine 48 hours prior to sacrifice. In all three experimental cases, ucn neurons were identified in the Edinger Westphal nucleus, superior olivary nucleus, and a few scattered neurons were found in the compact formation of the nucleus ambiguus (Ambc). However, in both female groups, ucn neurons were observed within the nucleus tractus solitarius (NTS) and the number of ucn neurons in the Ambc was increased compared to the male. Finally, in the OVX group distinct groups of ucn neurons were also identified in the rostral nucleus paragigantocellularis lateralis, the magnocellular reticular nucleus, and throughout the external formation of Amb. Taken together, these data suggest that the wide distribution of ucn neurons within autonomic brainstem structures in the OVX female may be associated with the greater effects observed on cardiovascular variables during stress. Heart and Stroke Foundation of Ontario

Mapping the rat somatosensory pathway from the anterior cruciate ligament nerve endings to the cerebrum

The anterior cruciate ligament (ACL) serves as the primary restraint to anterior tibial translation. In addition to this biomechanical function, the ACL appears to have a function in neuromuscular control. This hypothesis was formulated after the discovery of mechanoreceptors within the ACL. The full somatosensory pathway from the ACL to the cerebrum has yet to be elucidated. In order to map this sensory pathway, we conducted a viral trans-synaptic tracing experiment using the neurotropic pseudorabies virus (PRV). The pseudorabies virus was injected into the ACL of rats and allowed to replicate and spread trans-synaptically for 6-7 days. The brain and spinal cord of each sacrificed rat was then removed and processed immunohistochemically to detect the presence of PRV. PRV-immunoreactive neurons were found to be localized in several different regions from the spinal cord to the cerebrum. Four nuclei in the reticular formation of the brain stem demonstrated strong positive labeling: the mesencephalic reticular nucleus, magnocellular reticular nucleus, paragigantocellular reticular nucleus, and gigantocellular reticular nucleus. This finding suggests that the nerve endings of the rat ACL project into the cerebrum and that the reticular formation may play an important role in the afferent pathway of those nerve endings.

Mapping the rat somatosensory pathway from the anterior cruciate ligament nerve endings to the cerebrum

The anterior cruciate ligament (ACL) serves as the primary restraint to anterior tibial translation. In addition to this biomechanical function, the ACL appears to have a function in neuromuscular control. This hypothesis was formulated after the discovery of mechanoreceptors within the ACL. The full somatosensory pathway from the ACL to the cerebrum has yet to be elucidated. In order to map this sensory pathway, we conducted a viral trans-synaptic tracing experiment using the neurotropic pseudorabies virus (PRV). The pseudorabies virus was injected into the ACL of rats and allowed to replicate and spread trans-synaptically for 6–7 days. The brain and spinal cord of each sacrificed rat was then removed and processed immunohistochemically to detect the presence of PRV. PRV-immunoreactive neurons were found to be localized in several different regions from the spinal cord to the cerebrum. Four nuclei in the reticular formation of the brain stem demonstrated strong positive labeling: the mesencephalic reticular nucleus, magnocellular reticular nucleus, paragigantocellular reticular nucleus, and gigantocellular reticular nucleus. This finding suggests that the nerve endings of the rat ACL project into the cerebrum and that the reticular formation may play an important role in the afferent pathway of those nerve endings.

Neuroimmunological activation of the afferent laryngeal neural circuit in experimentally induced laryngeal inflammation

ConclusionsThese results show that laryngeal inflammatory reactions may induce the expression of proinflammatory cytokines along the afferent laryngeal circuit and in nuclei associated with the HPA axis. Local laryngeal inflammation may induce functional and physiologic alterations in the laryngeal neural system via neuroimmunologic reactions.ObjectiveIdiopathic laryngeal disorders associated with various neurologic conditions such as spasmodic dysphonia, idiopathic vocal fold paralysis and sudden infant death syndrome are causally related to upper respiratory tract infections, and it can be speculated that these disorders result in neurophysiologic alterations. The goal of this study was to identify the neurophysiologic effect on the central nervous system of local inflammatory alterations in the larynx.Material and methodsThe expression of c-fos and IL-1β was identified after injecting saline solution, 10 μg of lipopolysaccharide or 100 μg of lipopolysaccharide into the larynx of 12 rats.ResultsThe inflammatory cytokine IL-1β was mainly expressed in the inferior olivary nucleus and raphe nucleus, which are associated with the hypothalamic–pituitary–adrenal (HPA) axis. IL-1β expression was also found in the nuclei of afferent nervous pathways of the superior laryngeal nerve, such as the nucleus tractus solitarius, nucleus ambiguus, lateral reticular nucleus, magnocellular reticular nucleus and paragigantocellular reticular nucleus.

Démence fronto-temporale familiale avec inclusions marquées par l’anti-ubiquitine dans le tronc cérébral

Les démences fronto-temporales (DFT) forment un groupe hétérogène. Nous avons étudié une famille dont la mère et quatre enfants étaient atteints de DFT d’expression clinique typique, avec début précoce, avant 40 ans, par des troubles de la personnalité, et évolution prolongée, environ 30 ans, avec longue conservation de l’orientation spatiale, sans signe d’atteinte du motoneurone ni signes extrapyramidaux. L’étude neuropathologique d’un cas montra une atrophie massive prédominant au niveau des lobes frontaux. Dans les zones atrophiques, il existait une perte neuronale et un aspect spongieux des couches superficielles. Il n’y avait pas d’inclusion argentophile « de Pick », mais des inclusions marquées par l’anti-ubiquitine, mais non par l’anti-Tau, dans le fascia dentata. Dans le cytoplasme de certains neurones, dans les couches profondes du cortex mais surtout dans le tronc cérébral (réticulée magnocellulaire, noyaux du pont, noyaux moteurs, particulièrement noyau masticateur), de grandes inclusions argentophiles ovoïdes ou réniformes, peu denses, marquées seulement par l’anti-ubiquitine étaient observées. L’étude génétique mettait en évidence un Lod score positif non significatif avec des marqueurs flanquant le gène MAPτ, en l’absence de mutation décelable de ce gène. L’étude biochimique de la protéine Tau ne montrait pas d’anomalie qualitative. L’affection transmise dans la famille que nous avons étudiée paraît distincte aussi bien des DFT associées à une sclérose latérale amyotrophique (SLA) que des « taupathies ». Fronto-temporal dementias (FTD) were described a century ago on the macroscopic basis of frontal and/or temporal lobe atrophy. Progress in neuropathology, immunohistochemistry, biochemistry and genetics has since shown that they are heterogeneous entities, encompassing many different diseases with similar clinical presentations. A few, such as tauopathies due to mutations of the gene coding for tau protein (MAPτ form a well-defined group. Definition and grouping of other types of FTD is still problematic. We studied a family where the mother and 4/8 children were affected with FTD. Clinical presentation was typical of FTD. Onset was ill-defined with early (at age 40 years or less) personality changes. The clinical course was protracted (about 30 years). For a long period, the patients were able to live in the community in spite of obvious signs such as hyperorality and loss of verbal initiative; operative orientation as to place was preserved for a long time: a mute patient was still able to drive. Signs of extrapyramidal or motoneuron involvement were not observed. The genetic study failed to detect any mutation in MAPτ; the lod score for flanking markers was positive but not significant. Biochemical study showed no qualitative abnormality in tau protein. Neuropathological study of one affected subject showed brain atrophy (962g), with elective frontal lobe involvement. Cortical nerve cell loss was more marked in superficial layers and in frontal areas; glia was inconspicuous; pseudolaminar spongiosis was present in the more severely affected zones. No argentophilic “Pick bodies“ were seen; ubiquitin-positive, tau-negative round inclusions were present in the cytoplasm of fascia dentata neurones. “Tangles“ were mostly restricted to the entorhinal cortex, partly correlated with tau immunoreactivity, but better with ubiquitin immunoreactivity. Large, ovoid or reniform, moderately dense, spongy, granular or filamentous argentophilic cytoplasmic nerve cell inclusions were observed. They were ubiquitin-positive, but did not react with other antibodies, particularly anti-tau. They were present in swollen nerve cells in the deeper cortical layers but were most conspicuous in the brain stem: in the magnocellular reticular nuclei (e.g. nucleus centralis pontis ), in the pes pontis , in the inferior olive and in motor nuclei, especially in the trigeminal motor nucleus. They were not associated with nerve cell loss, atrophy nor pycnosis. Cerebellar relay nuclei neurones were swollen, and their cytoplasm contained argentophilic filaments. In our opinion, “ubiquitinopathy” would be non-specific and “Motor Neuron Disease-Inclusion Dementia” (MNDID) would not be satisfactory as a diagnosis for the present cases of FTD. Hopefully, progress in genetics may allow a causal, and thence definitive, classification.

Role for raphe magnus neuronal responses in the behavioral reactions to colorectal distension.

The brain stem is necessary for the expression of behavioral reactions to noxious visceral inputs. Neurons in raphe magnus (RM) and the adjacent nucleus reticularis magnocellularis (NRMC) respond to visceral stimuli and can facilitate the behavioral reaction to visceral stimulation. To determine which RM and NRMC cells could play a role in generating the reaction to colorectal distension (CRD), the responses of RM and NRMC cells to multiple intensities of CRD were compared with simultaneously evoked cardiovascular and visceromotor reactions in halothane-anesthetized rats. Most neurons (89%) responded to CRD with one of three basic response patterns. For cells with a graded response pattern, the response magnitude increased with increasing stimulation intensity. For flat responding cells, the response magnitude was not different across suprathreshold stimulation intensities. Finally, neurons with a switch response pattern responded to low- and high-intensity CRD in opposing directions. Cells were either inhibited or excited by CRD in each of these categories. Responses of cells with both graded and switch response patterns were significantly correlated with CRD-evoked tachycardia, pressor reaction, and hunching. The activity of graded-responding cells have the greatest predictive value for CRD-evoked reactions. Flat-responding cells have nonlinear responses that may augment reactions to stimuli above the noxious threshold. Cells with switch type response patterns may contribute to differential reactions evoked by CRD stimuli within the noxious range. In sum, RM and NRMC neurons respond to CRD with a variety of patterns, each of which may contribute to the sculpting of CRD reactions in different ways.

Cardioacceleratory responses to hypocretin-1 injections into rostral ventromedial medulla

Intracisternal injections of hypocretin-1 (hcrt-1) have been shown to elicit sympathoexciatory responses. However, the location of central sites that may mediate these cardiovascular effects have not been clearly elucidated. This study was done in male Wistar rats to investigate the effects of microinjections of hcrt-1 into the rostral ventromedial medulla (RVMM) on mean arterial pressure (MAP), heart rate (HR) and the arterial baroreflex. An initial series of experiments was done to provide a detailed mapping of the location of hcrt-1- and hcrt-1 receptors (hcrtR-1)-like immunoreactivity (ir) in the RVMM region. Hcrt-1 and hcrtR-1 ir were found throughout the RVMM region, but primarily within the magnocellular reticular nucleus and the adjacent nucleus paragigantocellularis lateralis. In the second series, this region containing hcrt-1 and hcrtR-1 ir was explored for sites that elicited changes in MAP and HR in the anaesthetized rat. Microinjection of hcrt-1 (0.5–2.5 pmol) into the region of magnocellular reticular nucleus elicited a dose-dependent increase in HR, with little or no change in MAP. Administration (iv) of the muscarinic receptor antagonist atropine methyl bromide significantly attenuated (∼62%) the HR response whereas, the total autonomic blockade abolished the HR response. Finally, unilateral or bilateral microinjection of hcrt-1 into the magnocellular reticular nucleus significantly attenuated the reflex bradycardia resulting from the activation of the baroreflex following the increase in MAP from an iv injection of phenylephrine. These data suggest that hcrt-1 in the RVMM region activates neuronal circuits that both inhibit vagal activity and increase sympathetic activity to the heart, and that it alters the excitability of central circuits that reflexly control the circulation.

Projections from the nucleus reticularis magnocellularis to the rat cervical cord using electrical stimulation and iontophoretic injection methods.

The aim of this study is to clarify the fiber distribution of the nucleus reticularis magnocellularis (NRMC) and adjacent areas in the rat spinal cord. Biotinylated dextran amine was injected iontophoretically through a glass capillary into the areas, in which a single cell responded to noxious electrical stimulation of the sciatic nerve and to a pinch of the thigh skin with multiple spikes. Labeled fibers descended bilaterally through the ventral funiculi of the medulla oblongata and then through the ventral and lateral funiculi of the cervical cord with an ipsilateral predominance, and terminated in the spinal gray (laminae I-X). A single fiber sometimes ran through several laminae while bifurcating many short branches with axon varicosities and terminal buttons in one transverse section, that is, through laminae V, VII and X, through laminae V, IIl-IV and I-II, and through laminae VII to I-II. The present study showed that the wide distribution of a single fiber and a mass of fibers descending from the NRMC and adjacent areas might modulate not only somatic sensory and motor functions but also autonomic functions in the spinal cord.

Raphe Magnus Neurons Respond to Noxious Colorectal Distension

Physiological studies of neurons in raphe magnus (RM) and the adjacent nucleus reticularis magnocellularis (NRMC) have demonstrated that the response to noxious cutaneous stimulation predicts the response to opioid administration and therefore a cell's functional role in nociceptive modulation. Although visceral stimulation, like opioids, elicits antinociception, little is known about how RM and NRMC cells respond to visceral stimulation. Therefore RM and NRMC cells were tested for their responses to both colorectal distension (CRD) and noxious cutaneous heat in halothane-anesthetized rats. Less than a third of serotonergic cells responded to CRD with small increases or decreases in discharge rate. In contrast, almost two-thirds of nonserotonergic cells responded to CRD stimulation with either excitatory (35%) or inhibitory (30%) responses to CRD. The response to heat did not predict the response to CRD with nearly equal proportions of heat-excited, -inhibited, and -unaffected cells being excited, inhibited, or unaffected by CRD. The dissociation between the responses to cutaneous heat and CRD demonstrates that cell classes based on the response to noxious heat are not homogeneous and may play multiple functional roles.

Physiological and anatomic evidence for functional subclasses of serotonergic raphe magnus cells.

Serotonergic cells in the medullary nucleus raphe magnus (RM) and adjacent nucleus reticularis magnocellularis (NRMC) project to the spinal cord where they are likely to modulate nociceptive transmission. Previous studies have suggested that these cells are physiologically and anatomically heterogeneous. In the present investigation, we examined whether subclasses of serotonergic RM and NRMC cells can be delineated based on their response to a visceral stimulus, and whether any such subclasses are morphologically distinct. Most RM and NRMC serotonergic cells tested (81 of 116) responded to retraction of the descending aorta into a polyethylene tube (the snare stimulus) with 57% of all cells tested excited and 13% inhibited. Responses of serotonergic cells to the snare outlasted the stimulus, were not reflective of evoked cardiovascular changes, and were observed in sino-aortic deafferented rats, evidence that the snare stimulus does not influence serotonergic cell discharge through activation of baroreceptors. Because serotonergic cells responsive to the snare were also responsive to mechanical brushing within the retroperitoneum, the snare is likely to change serotonergic cell discharge by means of the activation of mechanosensitive visceral afferents. Intracellular labeling of physiologically characterized serotonergic RM and NRMC cells showed that cells that were responsive to the snare stimulus had simpler axonal collateralization patterns than cells that were unresponsive to the snare stimulus. This association between morphological and physiological properties provides additional evidence that subpopulations of serotonergic cells exist and serve varied physiological functions.

Immunohistochemical localization of amylin in rat brainstem☆,☆☆

Immunohistochemical studies were conducted on rat brainstem using a specific polyclonal antiserum against the COOH-terminal (25–37) of human amylin. Amylin-immunoreactive cell bodies were observed in the vestibular, cochlear, trapezoid, and inner cerebellar nuclei and in the mesencephalic nucleus of trigeminal nerve. Positive cell bodies were also found in lateral, gigantocellular and magnocellular reticular nuclei. Numerous amylin-immunoreactive nerve fibers were shown in the trigeminal spinal tract, in the solitary area and in the area postrema. Amylin-immunoreactive cell bodies were often surrounded by a network of tyrosine hydroxylase-immunoreactive nerve fibers. These results provide morphologic evidence that amylin may play a role in some discrete sensory functions.

Serotonergic Raphe magnus cells that respond to noxious tail heat are not ON or OFF cells.

Pharmacological studies have suggested that serotonergic cells in RM contribute to both the inhibition and facilitation of spinal nociceptive transmission. Physiological studies in the medullary nucleus raphe magnus (RM) and adjacent nucleus reticularis magnocellularis have identified putative nociceptive-inhibitory OFF cells and nociceptive-facilitatory neurons ON cells by their responses to noxious thermal stimulation. The present study was designed to determine 1) whether any serotonergic RM cells respond to noxious thermal stimulation and 2) whether noxious heat-responsive serotonergic cells should be classified as ON or OFF cells. Serotonergic cells (n = 150) were identified by physiological criteria in anesthetized rats; 30 of 32 cells tested contained serotonin immunoreactivity. Noxious tail heat elicited a neuronal response in less than a quarter of the serotonergic cells. Most serotonergic cells that responded to tail heat were excited (n = 25), while a small minority of the cells tested were inhibited (n = 8). The tail heat-evoked responses of serotonergic cells were small in magnitude, averaging five to eight spikes in 10 s. Excitatory responses rarely persisted for more than 10 s, while inhibitory responses rarely persisted for more than 20 s. The tail heat-evoked responses of serotonergic cells were compared to those of non-serotonergic cells (n = 186). Non-serotonergic cells that responded to noxious tail heat had significantly greater response magnitudes, averaging 75-95 spikes in 10 s, than heat-responsive serotonergic cells. In addition, most heat-responsive non-serotonergic cells responded for at least 30 s after stimulus onset. These results demonstrate that the tail heat-evoked responses of serotonergic RM cells are qualitatively and quantitatively distinct from those of non-serotonergic ON and OFF cells. It is therefore unlikely that serotonergic RM cells, even the subpopulation that responds to noxious tail heat, share a physiological function with ON and OFF cells.

REM sleep behavior disorder

REM sleep is the stage associated with vivid dream mentation, desynchronous cortical EEG, and atonia of antigravitary muscles. REM sleep behavior disorder (RBD) is characterized by the intermittent loss of REM sleep atonia and by the appearance of elaborate motor activity associated with dream mentation. The animal model of REM sleep without atonia indicates that lesions to the perilocus coeruleus disrupt the excitatory connection to the nucleus reticularis magnocellularis in the descending medullary reticular formation and disable the hyperpolarization of the alpha spinal motoneurons. Extensive neurologic evaluations in humans suffering from both idiopathic and symptomatic forms have not identified specific lesions; however, findings in some patients suggest that diffuse lesions of the hemispheres, bilateral thalamic abnormalities, or primary brain-stem lesions may result in the RBD. Symptomatic RBD cases are associated with several neurologic disorders such as dementia, cerebrovascular diseases, multiple sclerosis, brain-stem neoplasm. RBD has been often documented to precede or to co-occur with neurodegenerative disorders, such as dementia, Parkinson's disease and multiple system atrophy. Most importantly, RBD is readily diagnosable and treatable. Patients and their bed partners usually report immediate improvement in sleep-related motor behavior with small doses of clonazepam.

Spinal and brain circuits to motoneurons of the bulbospongiosus muscle: Retrograde transneuronal tracing with rabies virus

Retrograde transneuronal tracing with rabies virus from the left bulbospongiosus muscle (BS) was used to identify the neural circuits underlying its peripheral and central activation. Rats were killed at 2, 3, 4, and 5 days post-inoculation (p.i.). Rabies immunolabelling was combined with immunohistochemical detection of choline acetyltransferase and oxytocin. Virus uptake was restricted to ipsilateral BS motoneurons (2 days p.i.). The onset of transfer (3 days p.i.) visualized interneurons in the dorsal grey commissure (DGC), intermediate zone, and sacral parasympathetic nucleus (SPN), mainly in DGC at L5–S1, and revealed synaptic connections between BS and external urethral sphincter motoneurons. At 4 and 5 days p.i., higher-order interneurons were labelled in other spinal areas and segments. Supraspinal labelling initially involved only Barrington's nucleus, nucleus reticularis magnocellularis, and paragigantocellularis lateralis (4 days p.i.). Later, labelling extended to other populations traditionally associated with control of sexual activity and micturition (periaqueductal grey, paraventricular nucleus, medial preoptic area, prefrontal cortex), but also indicated the intervention of somatic descending motor pathways (vestibulospinal and reticulospinal neurons, “hindlimb” regions of sensorimotor cortex and red nucleus) and cerebellar nuclei in multisynaptic innervation of the labelled motoneurons. Dual color immunofluorescence disclosed multisynaptic links between these motoneurons and thoracolumbar medial sympathetic (choline acetyltransferase-immunoreactive) neurons. In contrast, preganglionic neurons in SPN and most oxytocinergic neurons in paraventricular hypothalamic nucleus remained unlabelled, suggesting that parasympathetic and somatic outflow to pelvic organs are probably controlled by separate interneuronal populations and that oxytocinergic spinal projections are more likely to influence sacral autonomic rather than somatic outflow. J. Comp. Neurol. 414:167–192, 1999. © 1999 Wiley-Liss, Inc.

Spinal and brain circuits to motoneurons of the bulbospongiosus muscle: Retrograde transneuronal tracing with rabies virus

Retrograde transneuronal tracing with rabies virus from the left bulbospongiosus muscle (BS) was used to identify the neural circuits underlying its peripheral and central activation. Rats were killed at 2, 3, 4, and 5 days post-inoculation (p.i.). Rabies immunolabelling was combined with immunohistochemical detection of choline acetyltransferase and oxytocin. Virus uptake was restricted to ipsilateral BS motoneurons (2 days p.i.). The onset of transfer (3 days p.i.) visualized interneurons in the dorsal grey commissure (DGC), intermediate zone, and sacral parasympathetic nucleus (SPN), mainly in DGC at L5-S1, and revealed synaptic connections between BS and external urethral sphincter motoneurons. At 4 and 5 days p.i., higher-order interneurons were labelled in other spinal areas and segments. Supraspinal labelling initially involved only Barrington's nucleus, nucleus reticularis magnocellularis, and paragigantocellularis lateralis (4 days p.i.). Later, labelling extended to other populations traditionally associated with control of sexual activity and micturition (periaqueductal grey, paraventricular nucleus, medial preoptic area, prefrontal cortex), but also indicated the intervention of somatic descending motor pathways (vestibulospinal and reticulospinal neurons, "hindlimb" regions of sensorimotor cortex and red nucleus) and cerebellar nuclei in multisynaptic innervation of the labelled motoneurons. Dual color immunofluorescence disclosed multisynaptic links between these motoneurons and thoracolumbar medial sympathetic (choline acetyltransferase-immunoreactive) neurons. In contrast, preganglionic neurons in SPN and most oxytocinergic neurons in paraventricular hypothalamic nucleus remained unlabelled, suggesting that parasympathetic and somatic outflow to pelvic organs are probably controlled by separate interneuronal populations and that oxytocinergic spinal projections are more likely to influence sacral autonomic rather than somatic outflow.

Physiological survey of medullary raphe and magnocellular reticular neurons in the anesthetized rat.

The present study was designed to provide a detailed and quantitative description of the physiological characteristics of neurons in the medullary raphe magnus (RM) and adjacent nucleus reticularis magnocellularis (NRMC) under anesthetized conditions. The background discharge and noxious stimulus-evoked responses of RM and NRMC neurons were recorded in rats lightly anesthetized with isoflurane. All cells that were isolated successfully were studied. After recording background discharge, the neuronal response to repeated noxious thermal and noxious mechanical stimulation of the tail was recorded. Most cells were identified as nonserotonergic by their irregular or rapid background discharge pattern. Because the spontaneous discharge of most RM nonserotonergic cells contained pauses and bursts, a comparison between the change in rate evoked by tail heat and the range of rate changes that occur spontaneously was used to classify cells. The mean responses of ON and OFF cells were more than four times the standard deviation of the changes in rate observed spontaneously. ON cells were excited in 86% of the tail heat trials tested. Similarly, OFF cells were inhibited in 97% of the noxious tail heat trials tested. The heat-evoked changes in ON and OFF cell discharge varied over more than two orders of magnitude and were greater in cells with greater rates of background discharge. The heat-evoked responses of and cells had durations of tens of seconds to minutes and were always sustained beyond the visible motor response. Most ON and OFF cells responded to noxious tail clamp in a manner that was similar to their response to noxious heat. More than half of the NEUTRAL cells that were unresponsive to noxious heat were responsive to noxious tail clamp. A minority of ON, OFF, and NEUTRAL cells responded to innocuous brush stimulation with weak, transient responses. Although many cells discharged too infrequently to be classified, units with physiological properties that were different from those described above were rare. In conclusion, most RM and NRMC cells belong to three nonserotonergic physiological cell classes that can be distinguished from each other by the consistency, not the magnitude, of their responses to repeated noxious thermal stimulation. Because most of the heat-evoked change in and cell discharge occurs after the conclusion of the initial motor withdrawal, ON and OFF cells are likely to principally modulate the response to subsequent noxious insults.