Bulbar Reticular Formation Research Articles

The Ascending Reticular Activating System.

Discovery of the ascending reticular activating system (ARAS) can be attributed to work done in research neuroscientist Horace Magoun's laboratory. Before this finding, most scientists would focus on the diencephalon (and anterior midbrain) but not more caudally. Stimulation of the medial bulbar reticular formation in the pontine and midbrain tegmentum resulted disappearance of synchronized discharge and low-voltage fast activity. The effects were mediated by a thalamic projection system. This finding was a dramatic departure from the early philosophers' ascription of the awake soul to the ventricles (Galen), lumbosacral cord (Plato), pineal gland (Descartes), and even from more modern nineteenth- and twentieth-century hypotheses that the corpus striatum or periaqueductal gray matter housed the "seat of awareness." Magoun and his collaborators closed in on its true location in the cephalic brainstem-clinicians and neuropathologists would soon follow.

Brain cholinergic alterations in idiopathic REM sleep behaviour disorder: a PET imaging study with 18F-FEOBV

BackgroundREM sleep behaviour disorder (RBD) occurs frequently in patients with synucleinopathies such as Parkinson's disease, dementia with Lewy body, or multiple system atrophy, but may also occur as a prodromal stage of those diseases; and is termed idiopathic RBD (iRBD) when not accompanied by other symptoms. Cholinergic degeneration of the mesopontine nuclei have been described in synucleinopathies with or without RBD, but this has not yet been explored in iRBD. We sought to assess cholinergic neuronal integrity in iRBD using PET neuroimaging with the 18F-fluoroethoxybenzovesamicol (FEOBV). MethodsThe sample included 10 participants evenly divided between healthy subjects and patients with iRBD. Polysomnography and PET imaging with FEOBV were performed in all participants. Standardized uptake value ratios (SUVRs) were compared between the two groups using voxel wise t-tests. Non-parametric correlations were also computed in patients with iRBD between FEOBV uptake and muscle tonic and phasic activity during REM sleep. ResultsCompared with healthy participants, significantly higher FEOBV uptakes were observed in patients with iRBD. The largest differences were observed in specific brainstem areas corresponding to the bulbar reticular formation, pontine coeruleus/subcoeruleus complex, tegmental periacqueductal grey, and mesopontine cholinergic nuclei. FEOBV uptake in iRBD was also higher than in controls in the ventromedial area of the thalamus, deep cerebellar nuclei, and some cortical territories (including the paracentral lobule, anterior cingulate, and orbitofrontal cortex). Significant correlation was found between muscle activity during REM sleep, and SUVR increases in both the mesopontine area and paracentral cortex. ConclusionWe showed here for the first time the brain cholinergic alterations in patients with iRBD. As opposed to the cholinergic depletion described previously in RBD associated with clinical Parkinson's disease, increased cholinergic innervation was found in multiple areas in iRBD. The most significant changes were observed in brainstem areas containing structures involved in the promotion of REM sleep and muscle atonia. This suggests that iRBD might be a clinical condition in which compensatory cholinergic upregulation in those areas occurs in association with the initial phases of a neurodegenerative process leading to a clinically observable synucleinopathy.

Revealing the cerebello-ponto-hypothalamic pathway in the human brain

The cerebellum is shown to be involved in some limbic functions of the human brain such as emotion and affect. The major connection of the cerebellum with the limbic system is known to be through the cerebello-hypothalamic pathways. The consensus is that the projections from the cerebellar nuclei to the limbic system, and particularly the hypothalamus, or from the hypothalamus to the cerebellar nuclei, are through multisynaptic pathways in the bulbar reticular formation. The detailed anatomy of the pathways responsible for mediating these responses, however, is yet to be determined. Diffusion tensor imaging may be helpful in better visualizing the surgical anatomy of the cerebello-ponto-hypothalamic (CPH) pathway. This study aimed to investigate the utility of high-spatial-resolution diffusion tensor tractography for mapping the trajectory of the CPH tract in the human brain. Fifteen healthy adults were studied. We delineated, for the first time, the detailed trajectory of the CPH tract of the human brain in fifteen normal adult subjects using high-spatial-resolution diffusion tensor tractography. We further revealed the close relationship of the CPH tract with the optic tract, temporo-pontine tract, amygdalofugal tract and the fornix in the human brain.

Functional properties of nociceptive neurons in the nucleus centralis lateralis of the cat thalamus.

We studied functional properties of nociceptive neurons in the thalamic intralaminar nuclei, especially the nucleus centralis lateralis (CL) in urethane-chloralose anesthetized cats. Nociceptive neurons were found in the intralaminar nuclei, i.e. CL, nucleus centralis medialis (CeM) and nucleus parafascicularis (Pf). One third of these nociceptive neurons had their receptive fields throughout the body, including the orofacial area. Namely, they received convergent nociceptive input from the trigeminal and spinal nerve territories. These neurons with widely complex receptive fields were similar to those we found in the middle third of the caudal bulbar reticular formation (RF). Electrical stimulation of the CL antidromically activated about half of the caudal bulbar RF neurons tested. Furthermore, electrical stimulation of the cingulate gyrus antidromically activated some of CL neurons. These findings suggest that most of the CL neurons receiving nociceptive input from throughout the body receive information directly or indirectly via the caudal bulbar RF, and that certain of this information is then conveyed to the cingulate gyrus.

P95. REM sleep without atonia and REM sleep behavioral disorder in patients with amytrophic lateral sclerosis

REM sleep behavioral disorder (RBD) is characterized by dream-enacting behavior arising from REM sleep. On polysomnography (PSG), physiological muscle atonia is absent during phasic REM sleep in patients with RBD. RBD reflects disinhibition of cortical motor impulses on the brainstem level during REM sleep. RBD comprises a prodromal or concomitant condition in neurodegenerative diseases characterized by α-synuclein deposition. In patients with amyotrophic lateral sclerosis (ALS) RBD has not been systematically been investigated. Since neurodegeneration in ALS spreads from the motor cortex to the bulbar reticular formation we retrospectively studied the presence of REM sleep without atonia (RSWA) and RBD in patients with ALS. Patients with possible, probable or definite ALS according to the revised El Escorial criteria were included in the study. Occurrence of nocturnal behavioral abnormalities was assessed by a standardized physician interview. Exlusion criteria were tetraplegia, home ventilatory support, continous positive airway pressure support for obstructive sleep apnea, concomitant PD or MSA, dementia, and medication with either benzodiazepines or tricyclic antidepressants. Video polysomnography was performed according to standard recommendations, and PSG records were validated manually by two independent scorers according to the AASM criteria. Tonic muscle activity during REM sleep was scored when 50% ore more of a single 30 s epoch showed an EMG amplitude higher than the smallest EMG amplitude during non-REM sleep. Phasic muscle activity during REM sleep was scored when 5 out of 10 3 s mini-epochs showed transient muscle activity for 0.5–1.0 s with an EMG amplitude at least 4-fold above background activity. Classification of behavioral abnormalities during REM sleep was performed based on the widely used RBD severity scale (RBD-SS). Video-polysomnography records of 12 patients underwent detailed analysis. Three patients were female. Mean age was 48.8 years. One patient was diagnosed to have familial ALS with a proven mutation in the FUS gene. Two patients had bulbar onset of ALS symptoms. Behavioral abnormalities or increased motor acitivity during sleep clearly not associated with nocturnal dyspnea were reported by two patients and their bed partners. Subtle clinical signs of parkinsonism were present in only one of these patients on admission (mild hyposmia, distal rigor). Both patients showed dream-enacting behavior in combination with RSWA on video-PSG confirming RBD. Four patients without self-reported symptoms of possible RBD exhibited non-physiologic motor activity during REM sleep including jerks and vocalisations. Violent behavior or vigorous movements did not occur. 9 out of 12 patients showed tonic or phasic musle activity on surface EMG reflecting RSWA. It is not known whether ALS predisposes to the development of RBD. In accordance to very few case reports in the literature our study suggests that both RSWA and mild RBD may be encountered in patients with ALS. Further studies in larger patient samples are necessary to elucidate the prevalence of RBD in patients with ALS.

Mapping of Neurotensin in the Alpaca (Lama pacos) Brainstem

We studied the distribution of cell bodies and fibres containing neurotensin (NT) in the brainstem of the alpaca using an indirect immunoperoxidase technique. Immunoreactive fibres were widely distributed throughout the brainstem, whereas the distribution of cell bodies was less widespread. Immunoreactive perikarya were only found in the mesencephalic and bulbar reticular formation, periaqueductal grey, nucleus of the solitary tract, laminar spinal trigeminal nucleus and in the inferior colliculus. A high density of fibres containing NT was found in the dorsal nucleus of the raphe, marginal nucleus of the brachium conjunctivum, locus coeruleus, inferior colliculus, inter-peduncular nucleus, substantia nigra, periaqueductal grey, reticular formation of the mesencephalon, pons and medulla oblongata, nucleus of the solitary tract, laminar spinal trigeminal nucleus, hypoglossal nucleus, inferior central nucleus and in the tegmental reticular nucleus. The widespread distribution indicates that NT might be involved in multiple physiological actions in the alpaca brainstem; this must be investigated in the future as alpacas lives from 0 m above sea level to altitudes of up 5000 m and hence the involvement of this neuropeptide in special and unique regulatory physiological mechanisms could be suggested.

Projections of the hand field of the macaque ventral premotor area F5 to the brainstem and spinal cord

In the present study we first assessed that the hand motor field of the macaque ventral premotor area F5, involved in visuomotor control of hand actions, is connected to both the hand field of the primary motor cortex (M1) and the spinal cord. We then injected retroanterograde tracers in this field to completely illustrate its possible descending motor projections. In the brainstem the F5 hand motor field projects to the intermediate and deep layers of the superior colliculus (SC) and to sectors of the mesencephalic, pontine, and bulbar reticular formation, which are the sources of spinal projections. In the spinal cord, labeled terminals were virtually all confined to the C2-T1 segments, mostly contralaterally. At C6-T1 levels the labeling was weaker and mostly clustered laterally in the intermediate zone. At C2-C5 levels, labeled terminals were much denser and diffusely distributed over the mid-dorsal part of the intermediate zone where a propriospinal system that directly controls hand muscle motoneurons and mediates commands for the control of dexterous finger movements is located (Isa et al. [2007] Physiology 22:145-152). Thus, the F5 hand motor field has a weaker direct access and a stronger indirect access to spinal segments where hand muscle motoneurons are located, suggesting a role of this field in the generation and control of hand movements not only at the M1 level, but also at the spinal cord level. These projections may represent the neural substrate for the F5 hand motor field's role in the recovery of manual dexterity after M1 lesions.

Patterns of primary afferent depolarization of segmental and ascending intraspinal collaterals of single joint afferents in the cat

We have examined in the anesthetized cat the threshold changes produced by sensory and supraspinal stimuli on intraspinal collaterals of single afferents from the posterior articular nerve (PAN). Forty-eight fibers were tested in the L3 segment, in or close to Clarke's column, and 70 fibers in the L6-L7 segments within the intermediate zone. Of these, 15 pairs of L3 and L6-L7 collaterals were from the same afferent. Antidromically activated fibers had conduction velocities between 23 and 74 m/s and peripheral thresholds between 1.1 and 4.7 times the threshold of the most excitable fibers (xT), most of them below 3 xT. PAN afferents were strongly depolarized by stimulation of muscle afferents and by cutaneous afferents, as well as by stimulation of the bulbar reticular formation and the midline raphe nuclei. Stimulation of muscle nerves (posterior biceps and semitendinosus, quadriceps) produced a larger PAD (primary afferent depolarization) in the L6-L7 than in the L3 terminations. Group II were more effective than group I muscle afferents. As with group I muscle afferents, the PAD elicited in PAN afferents by stimulation of muscle nerves could be inhibited by conditioning stimulation of cutaneous afferents. Stimulation of the cutaneous sural and superficial peroneal nerves increased the threshold of few terminations (i.e., produced primary afferent hyperpolarization, PAH) and reduced the threshold of many others, particularly of those tested in the L6-L7 segments. Yet, there was a substantial number of terminals where these conditioning stimuli had minor or no effects. Autogenetic stimulation of the PAN with trains of pulses increased the intraspinal threshold in 46% and reduced the threshold in 26% of fibers tested in the L6-L7 segments (no tests were made with trains of pulses on fibers ending in L3). These observations indicate that PAN afferents have a rather small autogenetic PAD, particularly if this is compared with the effects of heterogenetic stimulation. Therefore, the depression of the PAN intraspinal fields produced by autogenetic stimulation described by Rudomin et al. (Exp Brain Res DOI 10.1007/s00221-006-0600-x, 2006) may be ascribed to other mechanisms besides a GABAa PAD. It is suggested that the small or no autogenetic PAD displayed by the examined joint afferents prevents presynaptic filtering of their synaptic actions and preserves the original information generated in the periphery. This could be important for proper adjustment of limb position.

The differential effects of pelvic and vagal inputs on the supraspinal cystitis viscero(noci)ceptive-related axis

Recently the development of the cyclophosphamide (CP, 100 mg/kg/ip) model has added an important element to the study of neural activities accompanying cystitis genesis. CP cystitis genesis results in the dual activation of the pelvic and vagal sensory afferent systems, which in turn activate a supraspinal network comprising the ventrocaudal bulbar reticular formation (vcBRF), the sensory subdivisions of the dorsal vagal complex (DVC) and its subcortical telencephalic targets, the dorsolateral subdivision of the bed nucleus of the stria terminalis (BSTLd) and the nucleus centralis of the amygdala (CeL). Altogether these structures form the sensory neural axis of the CP cystitis. However, both clinical and experimental observations have given evidence that only the pelvic afferents are at the origin of the painful sensation and related behaviour. Because of this, and for a better understanding of the nervous network that subserves cystitis painful information, we sought to determine whether the structures that constitute the cystitis sensory neural pathway have the same reactivity depending on the origin of the sensory afferent inputs, either pelvic or vagal. Using c- fos expression, which permits quantitative analysis of neural activity, we have demonstrated that the supraspinal CP cystitis responding structures do not form an homogeneous population in terms of sources of inputs. Although all structures are predominantly driven by vagal inputs, only the vcBRF, the DVC and the BSTLd respond to pelvic inputs. Consequently, and by referring to clinical observations, we have concluded that, it is these three areas, excluding the CeL, which constitute the main framework of the supraspinal pain sensory neural pathway of CP-induced cystitis. The activation of the vagus nerve would more probably relate to the other side effects that accompany CP injections such as nausea and headache attacks.

Environmental influences on viscero(noci)ceptive brain activities: the effects of sheltering

Visceral disorders are always accompanied by pain and/or a sense of ill-being that entails people to isolate themselves both physically and socially. By analogy with what happens in human beings, we have transferred to the rat the question of whether a protective, dark and quiet environment would influence the brain activities induced by visceral chemically-induced (cyclophosphamide [CP], 100 mg/kg/ip) adverse conditions of life. CP is an antitumoral drug that induces severe side effects (cystitis, headache, nausea, photophobia, phonophobia) and produces a strong state of ill-being in human beings. Brain activities were quantified using the expression of the Fos protein, a molecular marker of neuronal activity. The results compare data from groups of paired animals having been offered a shelter or not. Data were collected 4 h after the injection of CP, i.e., when cystitis was fully developed. Sheltered and unsheltered groups did not differ in bladder pathology. Intentional sheltering was shown to attenuate the expression of the CP-related Fos-Li activity within the locus coeruleus (LC) without affecting that of the structures known preferentially to process nociceptive inputs of bladder origin (dorsal vagal complex, ventrocaudal bulbar reticular formation, nucleus centralis of amygdala, dorsolateral portion of bed nucleus of stria terminalis). The LC levels of tyrosine hydroxylase and galanin neuronal contents were not affected. The LC belongs to the emotional activation system and can respond to a wide range of somatosensory and viscerosensory stimuli. Our hypothesis is that the LC would be processing the nervous activities that accompany the sense of ill-being coming from adverse conditions of life, including visceral disorders, and that voluntary isolation, by reducing its activity, would enable animals to minimize their level of distress.

Cyclophosphamide cystitis as a model of visceral pain in rats: a c-fos and Krox-24 study at telencephalic levels, with a note on pituitary adenylate cyclase activating polypeptide (PACAP).

This article is the fifth of a series aimed at mapping brain activities as they result from the development of cyclophosphamide (CP) cystitis in behaving rats using c-fos and Krox-24 expression. The inactive hepatic metabolites of CP are metabolized in the kidney to produce acrolein, which generates cystitis. Data come from animals which were injected once i.p. with either 1 ml saline (sham) or 100 mg/kg CP in 1 ml saline under transient volatile anesthesia and which behaved freely for 1-4 h postinjection, 4 h being the minimum time for cystitis to completely develop. Survival times longer than 4 h were not studied owing to ethical considerations. The first 2 h postinjection cover a period of time over which inputs of multifactorial origin (stress and pain due to the intraperitoneal injection process, possible effects due to the presence of hepatic CP metabolites in blood, cystitis onset) interact in an indistinguishable way; the last 2 h are more cystitis specific as the other effects have vanished. Complete screening of telencephalic levels has been performed. These data complete previously published data at both spinal and subtelencephalic levels. Of all the telencephalic structures, only the bed nucleus of the stria terminalis in the dorsal part of its lateral division (BSTLd) and, to a lesser degree, the nucleus centralis of the amygdala, mostly in its caudal portion (cCeA), appeared to be significantly driven over the most specific cystitis period. Both of these structures had related, but not identical patterns of expression. They both reacted shortly after CP injection, but, while cCeA maintained its activity throughout cystitis development, BSTLd showed a rebound, reaching a peak value when cystitis was fully developed. Both of these areas are the only telencephalic areas to contain high PACAP38 immunoreactivity. This is evidence that, (1) both the BSTLd and cCeA could be the most rostral areas that visceronociceptive inflow would reach when cystitis genesis is under way, and (2) PACAP38 could be one of the neurochemical agents involved in telencephalic visceronociceptive processing. From our complete mapping of brain activities under a fully developed cystitis situation (4 h postinjection), it appears that the activities in BSTLd and cCeA are concomitant with those of both the dorsal vagal complex (DVC), paratrigeminal nucleus (PaT), and the ventrocaudal bulbar reticular formation (vcBRF) at brainstem levels, suggesting they all form the main part of the neural network that subserves the central processing of cystitis-related inputs, comprising pain and associated pseudoaffective responses. Both the DVC and BSTLd, which are the most powerfully driven areas, would be particularly important in such a way. The origin of these activities should be found in both vagal (as sensed through PaT activity) and spinal (pelvic) influences. This network profoundly differs from those reported for painful situations, either somatic or visceral, which controversally accompany positive cardiac inotropism.

Opioid peptide involvement in the bulbar inhibition of electrodermal activity in the cat

By analogy with supraspinal and spinal inhibitory controls of pain, it was hypothesized that an opioid mechanism could be involved in the bulbar inhibitory control of the electrodermal activity. This activity was evoked as skin potential responses on the footpads of 13 cats by the central tegmental field stimulation (control responses) and inhibited by the simultaneous stimulation of bulbar reticular formation (experimental responses). Then, naloxone, an opioid peptide antagonist, was injected intravenously or intrathecally and its effects were analyzed on both control and experimental responses Intravenous injections of naloxone increased significantly the amplitude of experimental responses from 6 to 12 min after the injection and had no effect on the amplitude of control responses. Intrathecal injections of naloxone induced significant increases of amplitude of experimental responses from 6 to 42 min after the injection. These results showed that a spinal opioid peptide link could be involved in bulbar inhibition mechanisms of electrodermal activity.

Organization and efferent connections of the archistriatum of the mallard, Anas platyrhynchos L.: an anterograde and retrograde tracing study.

The intratelencephalic and descending connections of the archistriatum of the mallard were studied using anterograde and retrograde tracers. Autoradiography after injections of [3H]-leucine served to visualize the intratelencephalic and extratelencephalic efferent connections of the archistriatum. Horseradish peroxidase (HRP), HRP-wheatgerm agglutinin, and fluorescent tracers were used to identify the precise origin of the projections to the various terminal fields found in the anterograde experiments. Four main regions can be recognized in the archistriatum of the mallard: (1) the rostral or anterior part that is a source of contralateral intratelencephalic projections, in particular to the contralateral archistriatum; (2) the dorsal intermediate archistriatum that is the origin of a large descending fiber system, the occipitomesencephalic tract, with projections to dorsal thalamic nuclei, the medial spiriform nucleus, the intercollicular nucleus, the deep tectum, parts of the mesencephalic and bulbar reticular formation, and the subnuclei of the descending trigeminal tract. There are no direct projections to motor nuclei. This part corresponds to the somatic sensorimotor part as defined by Zeier and Karten (1971, Brain Res. 31:313-326); it also contributes to the ipsilateral intratelencephalic connections and, to a lesser degree, to contralateral intratelencephalic connections. (3) The ventral intermediate archistriatum is another region that is also a source of intratelencephalic projections, in particular of those to the lobus parolfactorius. The most lateral zone sends fibers to the septal area. (4) The caudoventral intermediate and posterior archistriatum is another region that is a source of the projections to the hypothalamus and thus corresponds to the amygdaloid part of the archistriatum as defined by Zeier and Karten; it also contributes a modest component to the occipitomesencephalic tract. The different cell populations are not spatially separated, which makes it impossible to recognize distinct subnuclei within the four main regions of the archistriatum of the mallard.

Patterns of connectivity of spinal interneurons with single muscle afferents.

A technique was developed to measure, in the anesthetized and paralyzed cat under artificial ventilation, changes of excitability to intraspinal stimulation simultaneously in two different afferent fibers or in two collaterals of the same afferent fiber. Intraspinal stimulation reduced the threshold of single muscle afferent fibers ending in the intermediate nucleus. This effect was seen with strengths below those required to activate the afferent fiber tested (1.5-12 microA), occurred at a short latency (1.5-2.0 ms), reached a maximum between 15 and 30 ms, and lasted up to 100 ms. The effects produced by graded stimulation applied at the shortest conditioning-testing stimulus time intervals increased by fixed steps, suggesting recruitment of discrete elements, most likely of last-order interneurons mediating primary afferent depolarization (PAD). The short-latency increases in excitability produced by the weakest effective intraspinal stimuli were usually detected only in the collateral closest to the stimulating micropipette, indicating that the stimulated interneurons mediating PAD have spatially restricted actions. The short-latency PAD produced by intraspinal stimuli, as well as the PAD produced by stimulation of the posterior biceps and semitendinosus (PBSt) nerve or by stimulation of the bulbar reticular formation (RF), was depressed 19-30 min after the i.v. injection of 0.5 mg/kg of picrotoxin, suggesting that all these effects were mediated by GABAergic mechanisms. The PAD elicited by stimulation of muscle and/or cutaneous nerves was depressed following the i.v. injection of (-)-baclofen, whereas the PAD elicited in the same collateral by stimulation of the RF was baclofen-resistant. The short-latency PAD produced by intraspinal stimulation was not always depressed by i.v. injections of (-)-baclofen. Baclofen-sensitive and baclofen-resistant monosynaptic PADs could be produced in different collaterals of the same afferent fiber. The results suggest that the intraspinal terminals of single muscle afferents receive synapses from more than one PAD-mediating GABAergic interneuron and that a single last-order interneuron has synaptic connections with a restricted number of intraspinal terminals and/or collaterals of the same afferent fiber. In addition, they support the existence of separate subsets of last-order baclofen-sensitive and baclofen-resistant interneurons that respond predominantly to segmental and to descending inputs. It is suggested that the restricted nature of the PAD plays an important role in the central control of the synaptic effectiveness of group I muscle afferents.

Electrical stimulation of the cholinergic laterodorsal tegmental nucleus elicits scopolamine-sensitive excitatory postsynaptic potentials in medial pontine reticular formation neurons

A large and consistent body of data implicates mesopontine cholinergic neurons in the production of rapid eye movement sleep, and indicates that many rapid eye movement sleep events are mediated by activation of pontine reticular formation neurons. There is anatomical evidence for projections from the mesopontine cholinergic nuclei to the pontine reticular formation, but no study has shown that stimulation of this cholinergic zone produces excitatory postsynaptic potentials in pontine reticular formation neurons. In the present study, intracellular recordings were made from 168 pontine reticular formation neurons, identified by antidromic activation from the bulbar reticular formation and by neurobiotin intracellular labeling, in acutely anesthetized cats. The effects of single-pulse electrical stimulation of the laterordorsal tegmental nucleus portion of the ipsilateral mesopontine cholinergic zone were evaluated in these neurons. Under urethane anesthesia this stimulation produced, in 21 of 22 recorded neurons, long-latency excitatory postsynaptic potentials (mean = 3 ms), consistent with the conduction velocity of unmyelinated cholinergic fibers (measured conduction velocity was 2 m/s). This excitatory postsynaptic potential was virtually abolished by intravenous administration of the muscarinic cholinergic receptor blocker scopolamine ( n = 40 neurons), and by acute cuts separating the laterodorsal tegmental nucleus and the recorded neurons ( n = 40). In contrast, a short-latency excitatory postsynaptic potential (0.7–1.5 ms) was not reduced in amplitude by scopolamine and could still be elicited following acute transverse cuts. Unlike the longer-latency excitatory postsynaptic potential, its amplitude was not reduced by barbiturate anesthesia. These data, suggesting the presence of an excitatory, cholinergic laterodorsal tegmental nucleus projection to the pontine reticular formation, provide further support to other lines of evidence implicating mesopontine cholinergic neurons in the production of rapid eye movement sleep, and are compatible with a model of rapid eye movement sleep generation in which a key element is mesopontine cholinergic input depolarizing and increasing the excitability of reticular core neurons.

Differential distribution of three types of nociceptive neurons within the caudal bulbar reticular formation in the cat

Nociceptive neurons within the reticular formation (RF) caudal to the obex were studied. 197 units recorded from the lateral part of subnucleus reticularis ventralis had receptive fields in the head, 72 units recorded from the medial RF in the body, and 160 units recorded from the middle third of RF in the head and body. About half of the units tested were antidromically excited by stimulation of nucleus centralis lateralis.

Cyclophosphamide cystitis as a model of visceral pain in rats. A survey of hindbrain structures involved in visceroception and nociception using the expression of c-Fos and Krox-24 proteins.

The evoked expression of the immediate early gene-encoded proteins c-Fos and Krox-24 was used to study activation of hindbrain neurons as a function of the development of cyclophosphamide (CP) cystitis in behaving rats. CP-injected animals received a single dose of 100 mg/kg i.p. under transient volatile anesthesia and survived for 1 to 4 h in order to cover the whole postinjection period during which the disease develops. CP-injected groups included: (1) animals with minor simple chorionic edema, an early characteristic of inflammation (1 h postinjection); (2) animals with well-developed simple chorionic edema (2 h postinjection); (3) animals with mild inflammation (chorionic edema accompanied by epithelial cleavage; 3 h postinjection); and (4) animals with complete inflammation (4 h postinjection). In addition to onset of chorionic edema, the earliest postinjection period also included the general aspects of the nervous reaction consecutive to the injection process (handling, transient volatile anesthesia and postanesthesia awakening, abdominal pinprick, CP-blood circulating effects). Controls included both noninjected animals and saline-injected animals surviving for the same times as CP-injected ones. Quantitative results come from c-Fos expression. It has been shown that: (1) saline injection is a significant stimulus for only nucleus O and central gray pars alpha and nucleus medialis of the dorsal vagal complex; (2) all structures driven by CP injection (nucleus O and central gray pars alpha, locus coeruleus, Barrington's nucleus and parabrachial area mostly in its ventral and lateral subdivisions, dorsal vagal complex, ventrocaudal portion of lateral bulbar reticular formation) responded vigorously shortly after injection, but only two (dorsal vagal complex, ventrocaudal portion of lateral bulbar reticular formation) showed increased or renewed activity when cystitis completely developed, i.e., when noxious visceral inputs reached highest levels. Regarding the sequential activation of these structures in relation to postinjection time, evidence is given that: (1) a large variety of hindbrain structures are differentially involved in either the general reaction consecutive to the injection process or to various degrees of cystitis; (2) these structures extend from the brain-spinal cord to the pons-mesencephalon transitional junction levels; (3) the two structures most powerfully driven by visceronociceptive inputs are also the most caudal ones, being located at the brain-spinal cord junction level; and (4) the dorsal vagal complex could be the main hindbrain visceral pain center, with three particular subdivisions, the nucleus medialis, nucleus commissuralis, and ventralmost part of area postrema, being involved.

Segmental and supraspinal control of synaptic effectiveness of functionally identified muscle afferents in the cat.

The present investigation documents the patterns of primary afferent depolarization (PAD) of single, functionally identified muscle afferents from the medial gastrocnemius nerve in the intact, anesthetized cat. Classification of the impaled muscle afferents as from muscle spindles or from tendon organs was made according to several criteria, which comprised measurement of conduction velocity and electrical threshold of the peripheral axons, and the maximal frequency followed by the afferent fibers during vibration, as well as the changes in discharge frequency during longitudinal stretch, the projection of the afferent fiber to the motor pool, and, in unparalyzed preparations, the changes in afferent activity during a muscle twitch. In confirmation of a previous study, we found that most muscle spindle afferents (46.1-66.6%, depending on the combination of criteria utilized for receptor classification) had a type A PAD pattern. That is, they were depolarized by stimulation of group I fibers of the posterior biceps and semitendinosus (PBSt) nerve, but not by stimulation of cutaneous nerves (sural and superficial peroneus) or the bulbar reticular formation (RF), which in many cases inhibited the PBSt-induced PAD. In addition, we found a significant fraction of muscle spindle primaries that were depolarized by stimulation of group I PBSt fibers and also by stimulation of the bulbar RF. Stimulation of cutaneous nerves produced PAD in 9.1-31.2% of these fibers (type B PAD pattern) and no PAD in 8.2-15.4% (type C PAD pattern). In contrast to muscle spindle afferents, only the 7.7-15.4% of fibers from tendon organs had a type A PAD pattern, 23-46.1% had a type B and 50-61.5% a type C PAD pattern. These observations suggest that the neuronal circuitry involved in the control of the synaptic effectiveness of muscle spindles and tendon organs is subjected to excitatory as well as to inhibitory influences from cutaneous and reticulospinal fibers. As shown in the accompanying paper, the balance between excitation and inhibition is not fixed, but can be changed by crushing the afferent axons in the peripheral nerve and allowing subsequent reconnection of these afferent fibers with muscle receptors.

Changes in PAD patterns of group I muscle afferents after a peripheral nerve crush.

In the anesthetized cat we have analyzed the changes in primary afferent depolarization (PAD) evoked in single muscle spindle and tendon organ afferents at different times after their axons were crushed in the periphery and allowed to regenerate. Medial gastrocnemius (MG) afferents were depolarized by stimulation of group I fibers in the posterior biceps and semitendinosus nerve (PBSt), as soon as 2 weeks after crushing their axons in the periphery, in some cases before they could be activated by physiological stimulation of muscle receptors. Two to twelve weeks after crushing the MG nerve, stimulation of the PBSt produced PAD in all MG fibers reconnected with presumed muscle spindles and tendon organs. The mean amplitude of the PAD elicited in afferent fibers reconnected with muscle spindles was increased relative to values obtained from Ia fibers in intact (control) preparations, but remained essentially the same in fibers reconnected with tendon organs. Quite unexpectedly, we found that, between 2 and 12 weeks after crushing the MG nerve, stimulation of the bulbar reticular formation (RF) produced PAD in most afferent fibers reconnected with muscle spindle afferents. The mean amplitude of the PAD elicited in these fibers was significantly increased relative to the PAD elicited in muscle spindle afferents from intact preparations (from 0.08 +/- 0.4 to 0.47 +/- 0.34 mV). A substantial recovery was observed between 6 months and 2.5 years after the peripheral nerve injury. Stimulation of the sural (SU) nerve produced practically no PAD in muscle spindles from intact preparations, and this remained so in those afferents reconnected with muscle spindles impaled 2-12 weeks after the nerve crush. The mean amplitude of the PAD produced in afferent fibers reconnected with tendon organs by stimulation of the PBSt nerve and of the bulbar RF remained essentially the same as the PAD elicited in intact afferents. However, SU nerve stimulation produced a larger PAD in afferents reconnected with tendon organs 2-12 weeks after the nerve crush (mean PAD changed from 0.05 +/- 0.04 to 0.32 +/- 0.17 mV). The results obtained indicate that the PAD patterns of the afferent fibers reconnected with muscle spindle and tendon organ afferents are changed after crushing their axons in the periphery: stimulation of the bulbar RF appears to produce larger PAD in fibers reconnected with muscle spindles, and stimulation of cutaneous afferents produces larger PAD in fibers reconnected with tendon organs. It is suggested that these alterations in the patterns of PAD of muscle afferents result from central changes in the balance of excitatory and inhibitory influences acting on the segmental pathways mediating the PAD. Although the functional role of these changes has not been established, they may reflect compensatory changes aimed to adjust information arising from damaged afferents.

Raphe magnus and reticulospinal actions on primary afferent depolarization of group I muscle afferents in the cat.

1. In the anaesthetized cat, electrical stimulation of the bulbar reticular formation produced a short latency (2.1 +/- 0.3 ms) positive potential in the cord dorsum. In contrast, stimulation of the nucleus raphe magnus with strengths below 50 microA evoked a slow negative potential with a mean latency of 5.5 +/- 0.6 ms that persisted after sectioning the contralateral pyramid and was abolished by sectioning the ipsilateral dorsolateral funiculus. 2. The field potentials evoked by stimulation of the bulbar reticular formation and of the nucleus raphe magnus had a different intraspinal distribution, suggesting activation of different sets of segmental interneurones. 3. Stimulation of these two supraspinal nuclei produced primary afferent depolarization (PAD) in single Ib fibres and inhibited the PAD elicited by group I volleys in single Ia fibres. The inhibition of the PAD of Ia fibres produced by reticulospinal and raphespinal inputs appears to be exerted on different interneurones along the PAD pathway. 4. It is concluded that, although reticulospinal and raphespinal pathways have similar inhibitory effects on PAD of Ia fibres, and similar excitatory effects on the PAD of Ib fibres, their actions are conveyed by partly independent pathways. This would allow their separate involvement in the control of posture and movement.