Medullary Serotonergic Neurons Research Articles

Sudden Unexplained Death in Childhood: A Neuropathology Review.

Sudden Unexplained Death in Childhood (SUDC) is the unexpected death of a child over age 12 months that remains unexplained after a thorough case investigation, including review of the child's medical history, circumstances of death, a complete autopsy and ancillary testing (1). First defined in 2005, SUDC cases are more often male, with death occurring during a sleep period, being found prone, peak winter incidence, associated with febrile seizure history in ~28% of cases and mild pathologic changes insufficient to explain the death (1, 2). There has been little progress in understanding the causes of SUDC and no progress in prevention. Despite reductions in sudden unexpected infant death (SUID) and other causes of mortality in childhood, the rate of SUDC has increased during the past two decades (3–5). In Ireland, SUID deaths were cut in half from 1994 to 2008 while SUDC deaths more than doubled (4). Surveillance issues, including lack of standardized certification practices, affect our understanding of the true magnitude of unexplained child deaths. Mechanisms underlying SUDC, like SUID, remain largely speculative. Limited and inconsistent evidence implicates abnormalities in brainstem autonomic and serotonergic nuclei, critical for arousal, cardiorespiratory control, and reflex responses to life-threatening hypoxia or hypercarbia in sleep (6). Abnormalities in medullary serotonergic neurons and receptors, as well as cardiorespiratory brainstem nuclei occur in some SUID cases, but have never been studied in SUDC. Retrospective, small SUDC studies with non-standardized methodologies most often demonstrate minor hippocampal abnormalities, as well as focal cortical dysplasia and dysgenesis of the brainstem and cerebellum. The significance of these findings to SUDC pathogenesis remains unclear with some investigators and forensic pathologists labeling these findings as normal variants, or potential causes of SUDC. The development of preventive strategies will require a greater understanding of underlying mechanisms.

Comparison of intrinsic chemosensitivity of acutely dissociated medullary serotonergic and retrotrapezoid nucleus neurons

ObjectiveSerotonergic neurons of the medullary raphe and Phox2b expressing neurons of the retrotrapezoid nucleus (RTN) are both putative respiratory chemoreceptors. Here we compare their intrinsic response to acidosis using patch clamp recordings after acute dissociation in vitro.MethodsThe midline medulla and rostral ventrolateral medulla were dissected from mice (>P10) that expressed fluorescent reporters: 1) Phox2b‐Cre::Floxed‐tdTomato:ChAT‐GFP for RTN, and; 2) ePet‐EYFP for 5‐HT. Cells were treated with papain, mechanically dissociated, plated on CELLocate coverslips and allowed to adhere for a variable period before recordings were made. RTN neurons were identified as those that expressed tdTomato but not GFP. In some cases, immunohistochemistry was used to ensure that RTN neurons did not express tyrosine hydroxylase. Patch clamp recordings were made with standard methods previously used to study chemosensitivity of these neurons in culture and slices. Neurons were considered “chemosensitive” if they increased their firing rate by more than 20% in response to a decrease in pH from 7.4 to 7.2 (due to an increase in PCO2 from 5% to 9%, balance O2). Neurons were chosen for recordings only if they were isolated from other neurons. In some cases a lack of synaptic connections was verified with immunohistochemistry for MAP2 using confocal microscopy. The methods used to dissociate and record 5‐HT and RTN neurons were identical.Results45% of 5‐HT neurons (n=84) were chemosensitive, with a mean increase in firing rate by 135%+/−115% (mean+/−SD). 38% of these neurons increased firing rate by more than 50% (i.e. to 150% of baseline firing rate. In contrast, only 29% of RTN neurons (n=21) increased their firing rate by more than 20%, and none of them increased their firing rate by more than 50%. Chemosensitive RTN neurons increased firing rate by a mean of 29%+/−8%. The mean firing rate of chemosensitive 5‐HT neurons was 1.1+/−0.6 Hz at pH 7.4 and 2.15+/−0.9 Hz at pH 7.2 (n=38; p<0.0001). The mean firing rate of chemosensitive RTN neurons was 3.3+/−2.3 Hz at pH 7.4 and 4.3+/−2.8 Hz at pH 7.2 (n=6; p<0.05). Recordings were performed in a separate cohort of 5‐HT neurons using a change in CO2 from 5% to 9% in 21% O2 (balance N2) and their response was not different than in 95% O2.Conclusions5‐HT neurons had a high degree of intrinsic (cell‐autonomous) chemosensitivity to small changes in pH that was similar to that reported for these neurons in acute brain slices and in cell culture. Intrinsic chemosensitivity of RTN neurons was uncommon. When present it was small, similar to that previously reported in a small percentage of neurons in the neocortex, hippocampus and cerebellum. RTN neurons may play a more important role either as “emergency chemoreceptors” (i.e. responding to very large pH changes) or as relays from other chemoreceptors than as pH sensors themselves.Support or Funding InformationNIH U01 NS090414

Inactivation of Serotonergic Neurons in the Rostral Medullary Raphé Attenuates Stress-Induced Tachypnea and Tachycardia in Mice.

The medullary raphé nuclei are involved in controlling cardiovascular, respiratory, and thermoregulatory functions, as well as mediating stress-induced tachycardia and hyperthermia. Although the serotonergic system of the medullary raphé has been suggested as the responsible entity, specific evidence has been insufficient. In the present study, we tested this possibility by utilizing an optogenetic approach. We used genetically modified mice [tryptophan hydroxylase 2 (Tph2); archaerhodopsin-T (ArchT) mice] in which ArchT, a green light-driven neuronal silencer, was selectively expressed in serotonergic neurons under the regulation of Tph2 promoters. We first confirmed that an intruder stress selectively activated medullary, but not dorsal or median raphé serotonergic neurons. This activation was suppressed by photo-illumination via a pre-implanted optical fiber, as evidenced by the decrease of a cellular activation marker protein in the neurons. Next, we measured electro cardiogram (ECG), respiration, body temperature (BT), and locomotor activity in freely moving mice during intruder and cage-drop stress tests, with and without photo-illumination. In the intruder test, photo inactivation of the medullary serotonergic neurons significantly attenuated tachycardia (362 ± 58 vs. 564 ± 65 bpm.min, n = 19, p = 0.002) and tachypnea (94 ± 82 vs. 361 ± 138 cpm.min, n = 9, p = 0.026), but not hyperthermia (1.0 ± 0.1 vs. 1.0 ± 0.1°C.min, n = 19, p = 0.926) or hyperlocomotion (17 ± 4 vs. 22 ± 4, arbitrary, n = 19, p = 0.089). Similar results were obtained from cage-drop stress testing. Finally, photo-illumination did not affect the basal parameters of the resting condition. We conclude that a subpopulation of serotonergic neurons in the medullary raphé specifically mediate stress-induced tachypnea and tachycardia, which have little involvement in the basal determination of respiratory frequency (Res) and heart rate (HR), specifically mediate stress-induced tachycardia and tachypnea.

Non-motor multiple system atrophy associated with sudden death: pathological observations of autonomic nuclei

Multiple system atrophy (MSA) manifests as a combination of dysautonomia and motor symptoms/signs. However, rare cases presenting with autonomic failures in absence of motor symptoms/signs until their deaths have been reported and are referred to as non-motor MSA. To clarify pathological findings underlying non-motor MSA patients, we analyzed consecutively autopsied 161 patients with MSA. In results, four patients were identified as having non-motor MSA, who showed isolated autonomic disorders throughout their lives and had minimal pathological changes in the motor systems. We also identified two patients with pathologically minimal MSA, who had minimal pathological involvement in the motor systems and presented with definite parkinsonism and dysautonomia. Survival durations of the non-motor MSA patients were much shorter (1.3-2.0years) than those of the classical MSA patients (3.0-7.0years), and the causes of death were all sudden death. The medullary serotonergic neurons were severely involved in the non-motor MSA patients in comparison with the classical MSA patients. Also, one of the pathologically minimal MSA patients had died suddenly and exhibited marked involvement of the medullary serotonergic neurons. The involvement of the medullary catecholaminergic or cholinergic neurons did not differ in severities among the groups. We conclude that non-motor MSA may be a pathological variant of MSA that preferentially involves the medullary serotonergic neurons and autonomic systems in association with poor prognosis.

Impaired Serotonergic Brainstem Function during and after Seizures.

Seizures often cause impaired breathing, cardiac dysfunction, and loss of consciousness. The brainstem and, specifically, brainstem serotonin neurons are thought to play an important role in controlling breathing, cardiac function, and arousal. We used an established rat seizure model to study the overall neuronal activity in the brainstem as well as firing of specific serotonin neurons while measuring cardiorespiratory function. Our results demonstrated overall decreases in brainstem neuronal activity and marked downregulation of lower brainstem serotonin neuronal firing in association with decreased breathing and heart rate during and after seizures. These findings point the way toward new treatments to augment brainstem function and serotonin, aiming to prevent seizure complications and reduce morbidity and mortality in people living with epilepsy.

Medullary serotonergic system: organization, effects, and clinical correlations.

Medullary neurons synthesizing serotonin (5-HT) encompass several groups projecting to the spinal cord and brainstem. The medullary serotonergic groups include the nucleus raphe magnus (RMg), which projects primarily to the spinal and trigeminal dorsal horn, and the nucleus raphe obscurus (ROb) and raphe pallidus (RPa), which project to motor and autonomic nuclei of the spinal cord and medulla. Other serotonergic groups in the ventrolateral medullary reticular formation contribute bulbospinal and propriospinal projections to the spinal cord and brainstem. Serotonin exerts complex effects on its targets via several subtypes of 5-HT receptors. Medullary serotonergic neurons exert a major modulatory influence on nociceptive processing, motoneuron excitability, respiratory control, and autonomic function, particularly thermoregulatory sympathetic responses to cold. Not surprisingly, these neurons have been implicated in the pathophysiology of pain, spasticity, and respiratory failure potentially related to sudden death.

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.

Silencing vesicular neurotransmission in medullary serotonergic (5HT) neurons causes prolonged and failed autoresuscitation during early postnatal life in mice: possible implications for the sudden infant death syndrome (SIDS)

Abnormalities in the medullary 5HT system, including decreased tissue levels of 5HT, have been identified in approximately 70% of SIDS cases studied. Failed autoresuscitation (inability to recover breathing and heart rate following hypoxic apnea) is considered a possible cause of death in SIDS infants. Using 5HT ‘silenced’ mice, we examined the effect of altered neurotransmitter release from 5HT neurons on the ability to autoresuscitate after repeated exposure to intermittent anoxia at postnatal days (P) 5, 8 and 12. In these mice, Cre recombinase activates expression of tetanus toxin light chain in medullary 5HT neurons, which cleaves the VAMP2 protein that anchors vesicles to the synaptic membrane, thus suppressing synaptic vesicular neurotransmission in these neurons. ‘Silenced’ pups took twice as long as controls to initiate gasping at P5 and P8, and had a 22% reduction in gasp frequency at P8. Recovery of heart rate and breathing to 90% of normal was prolonged up to 5 and 2 times, respectively compared to controls at P8. Autoresuscitation failed in 19% of the ‘silenced’ pups at P5 and 44% at P8, compared to no control pup deaths. These data indicate that suppressing vesicular neurotransmission in 5HT neurons delays initiation of gasping and suggest that this may contribute to prolonged or failed autoresuscitation in response to anoxia in early postnatal life. (HD036379)

Depletion of medullary serotonergic neurons in patients with multiple system atrophy who succumbed to sudden death

Depletion of medullary serotonergic neurons in patients with multiple system atrophy who succumbed to sudden death

Depletion of medullary serotonergic neurons in patients with multiple system atrophy who succumbed to sudden death

Multiple system atrophy (MSA) is a neurodegenerative disorder characterized by prominent autonomic failure with ataxia and/or parkinsonism. The leading cause of death in MSA is sudden death. We have shown that the early development of autonomic failure is an independent risk factor for sudden death. The depletion of sympathetic preganglionic neurons in the spinal intermediolateral cell column (IML) and its afferent medullary catecholaminergic and serotonergic neurons has been proposed to be partly responsible for autonomic failure in MSA. In this study, we investigated whether the depletion of neurons in any of these autonomic neuron groups contributes to sudden death in MSA. Out of 52 autopsy-proven patients with MSA, we selected 12 individuals who had died within 3.5 years after disease onset to define the accurate levels of slices and identify early neuropathological changes of autonomic nuclei in MSA. Four patients succumbed to sudden death and eight patients died through established causes. Serial 10 mum sections were obtained from the 8th segment of the thoracic cord and the rostral medulla oblongata. Sections from the medulla oblongata were immunostained for thyrosine hydroxylase and tryptophan hydroxylase. The total cell number in the five sections was computed for comparison. Compared with the control, the MSA group showed a marked depletion of neurons in the IML (38.0 +/- 7.1 versus 75.2 +/- 7.6 cells, P < 0.001), thyrosine hydroxylase-immunoreactive neurons in the ventrolateral medulla (VLM) (17.4 +/- 5.1 versus 72.8 +/- 13.6 cells, P < 0.01) and tryptophan hydroxylase-immunoreactive neurons in the VLM (15.6 +/- 9.2 versus 60.8 +/- 17.0 cells, P < 0.01), nucleus raphe obscurus (19.3 +/- 4.4 versus 75.3 +/- 8.6 cells, P < 0.001), nucleus raphe pallidus (2.1 +/- 2.7 versus 9.0 +/- 3.4 cells, P < 0.03), and arcuate nucleus (0.4 +/- 0.8 versus 2.3 +/- 1.5 cells, P < 0.05). Moreover, in patients who succumbed to sudden death, when compared with patients who had established causes of death, we found a marked depletion of tryptophan hydroxylase-immunoreactive neurons in the VLM (7.3 +/- 3.5 versus 21.8 +/- 6.5 cells, P < 0.02) and nucleus raphe obscurus (15.0 +/- 2.0 versus 22.5 +/- 2.1 cells, P < 0.01). The results indicate that the spinal IML and medullary catecholaminergic and serotonergic systems are involved even in the early stages of MSA, and the dysfunction of the medullary serotonergic system regulating cardiovascular and respiratory systems could be responsible for sudden death in patients with MSA.

Serotonin, gasping, autoresuscitation, and SIDS—a contrarian view

in their study in the Journal of Applied Physiology , Erikson and Sposato ([4][1]) induced anoxic apnea in neonatal Pet -1 knockout (KO) and wild-type mice at postnatal day (P) 4.5 and P9 and examined the subsequent quality of gasping and autoresuscitation while the animals breathed room air. Pet-1

Serotonin transporter knockout mice have a reduced ventilatory response to hypercapnia (predominantly in males) but not to hypoxia

Medullary serotonergic (5-HT) neurons are implicated in central chemoreception and 5-HT abnormalities are present in many cases of the sudden infant death syndrome (SIDS). Mice with a targeted disruption of the serotonin transporter (5-HTT) develop in the presence of excess 5-HT in brain extracellular fluid (ECF). As adults they exhibit reduced 5-HT neuron activity and 5-HT1A receptor binding with varying changes in postsynaptic 5-HT receptor function. They exhibit behavioural phenotypes (anxiety, reduced aggression) but little is known about their control of breathing. We show that conscious adult male and female 5-HTT knockout mice breathing air at room temperature have a higher resting (.)VO2, breathing frequency and (.)VE but a normal body temperature and (.)VE/ (.)VO2 ratio (the ventilatory equivalent) compared to wild-type (WT) controls. In hypercapnia, there is a reduced ventilatory response (expressed as the (.)VE/ (.)VO2 ratio) that is much more prominent in males (-68%) than females (-22%). In hypoxia, both males and females exhibit a higher (.)VE, (.)VO2 and body temperature but their (.)VE/ (.)VO2 ratio is normal. We conclude that 5-HTT knockout mice have a diminished function of the medullary 5-HT system, which is manifest most remarkably in a substantial loss of CO2 sensitivity predominantly in males. This finding supports the importance of medullary 5-HT neurons in central chemoreception. Females either rely less on 5-HT neurons in chemoreception or adapt more readily to the loss of 5-HT function. This genetic model allows examination of the role of excess 5-HT in ECF in the development of the control of breathing and central chemoreception, which may be pertinent to SIDS.

Necdin Plays a Role in the Serotonergic Modulation of the Mouse Respiratory Network: Implication for Prader-Willi Syndrome

Prader-Willi syndrome is a neurogenetic disease resulting from the absence of paternal expression of several imprinted genes, including NECDIN. Prader-Willi children and adults have severe breathing defects with irregular rhythm, frequent sleep apneas, and blunted respiratory regulations. For the first time, we show that Prader-Willi infants have sleep apneas already present at birth. In parallel, in wild-type and Necdin-deficient mice, we studied the respiratory system with in vivo plethysmography, in vitro electrophysiology, and pharmacology. Because serotonin is known to contribute to CNS development and to affect maturation and function of the brainstem respiratory network, we also investigated the serotonergic system with HPLC, immunohistochemistry, Rabies virus tracing approaches, and primary culture experiments. We report first that Necdin-deficiency in mice induces central respiratory deficits reminiscent of Prader-Willi syndrome (irregular rhythm, frequent apneas, and blunted respiratory regulations), second that Necdin is expressed by medullary serotonergic neurons, and third that Necdin deficiency alters the serotonergic metabolism, the morphology of serotonin vesicles in medullary serotonergic neurons but not the number of these cells. We also show that Necdin deficiency in neonatal mice alters the serotonergic modulation of the respiratory rhythm generator. Thus, we propose that the lack of Necdin expression induces perinatal serotonergic alterations that affect the maturation and function of the respiratory network, inducing breathing deficits in mice and probably in Prader-Willi patients.

Ventilatory response to hypercapnia and hypoxia after extensive lesion of medullary serotonergic neurons in newborn conscious piglets

Acute inhibition of serotonergic (5-HT) neurons in the medullary raphé (MR) using a 5-HT(1A) receptor agonist had an age-dependent impact on the "CO(2) response" of piglets (33). Our present study explored the effect of chronic 5-HT neuron lesions in the MR and extra-raphé on the ventilatory response to hypercapnia and hypoxia in piglets, with possible implications on the role of 5-HT in the sudden infant death syndrome. We established four experimental groups. Group 1 (n = 11) did not undergo any treatment. Groups 2, 3, and 4 were injected with either vehicle or the neurotoxin 5,7-dihydroxytryptamine in the cisterna magna during the first week of life (group 2, n = 9; group 4, n = 11) or second week of life (group 3, n = 10). Ventilation was recorded in response to 5% CO(2) (all groups) and 12% O(2) (group 2) during wakefulness and sleep up to postnatal day 25. Surprisingly, the piglets did not reveal changes in their CO(2) sensitivity during early postnatal development. Overall, considerable lesions of 5-HT neurons (up to 65% decrease) in the MR and extra-raphé had no impact on the CO(2) response, regardless of injection time. Postlesion raphé plasticity could explain why we observed no effect. 5,7-Dihydroxytryptamine-treated males, however, did present a lower CO(2) response during sleep. Hypoxia significantly altered the frequency during sleep in lesioned piglets. Further studies are necessary to elucidate the role of plasticity, sex, and 5-HT abnormalities in sudden infant death syndrome.

Single‐unit responses of serotonergic medullary raphe neurons to cardiovascular challenges in freely moving cats

Single-unit activity of serotonergic neurons in the nuclei raphe obscurus (NRO) and raphe pallidus (NRP) were recorded in conjunction with heart rate in freely moving cats in response to systemic administration of vasoactive drugs and to graded haemorrhage. Bolus administration of phenylephrine hydrochloride and sodium nitroprusside (20 microg/kg, i.v.) produced a marked, transient reflex bradycardia (-42 b.p.m.) and tachycardia (+60 b.p.m.), respectively. The activity of NRO/NRP serotonergic neurons remained unchanged after phenylephrine and nitroprusside administration. The administration of hydralazine (1 mg/kg, i.v.), a long-acting vasodilator, produced sustained tachycardia (+60 b.p.m.), which was not accompanied by changes in neuronal activity, despite prolonged reflex activation of the sympathetic nervous system. The initial withdrawal of up to 15% of total blood volume increased heart rate (+12 b.p.m.), whereas the removal of 22.5% of total blood decreased heart rate (-44 b.p.m.). The activity of NRO/NRP serotonergic neurons remained unaltered throughout graded haemorrhage trials, despite the changes in sympathetic outflow. Thus, serotonergic NRO and NRP neurons appear to be insensitive to alterations in blood pressure and baroreceptor activity, and this lack of responsiveness does not support a specific role for these cells in cardiovascular regulation. Furthermore, these neurons do not appear to be involved in physiological mechanisms underlying alterations in autonomic outflow invoked by hypertension and hypotension. Taken within the context of our previous work, the present data suggest that medullary serotonergic neurons may modulate autonomic outflow, but only in relation to their primary role in motor control.

Insulin-induced hypoglycemia decreases single-unit activity of serotonergic medullary raphe neurons in freely moving cats: relationship to sympathetic and motor output.

Serotonergic single-unit activity during glucoregulatory challenges was studied in the nuclei raphe obscurus (NRO) and raphe pallidus (NRP) of freely moving cats. Systemic insulin administration (2-4 IU/kg, i.v.) suppressed neuronal activity by approximately 40% in direct relationship to blood glucose levels and in inverse relationship to plasma catecholamine levels. NRO and NRP serotonergic neurons displayed a temporary recovery in unit activity in response to i.v. glucose administration (500 mg/kg), which temporarily reversed insulin-induced hypoglycemia. The neuronal responses to insulin and subsequent glucose administration were also directly related to changes in integrated nuchal electromyographic activity. Serotonergic unit activity remained unchanged after glucose loading (500 mg/kg, i.v.), which produced a four-fold increase in blood glucose. Thus, medullary serotonergic neurons appear to be sensitive to reductions, but not increases, in blood glucose. The observed inverse relationship between unit activity and plasma catecholamines does not support a postulated sympathoexcitatory role for these neurons. Instead, the parallel changes in single-unit activity and integrated muscle activity support the hypothesis that the activity of medullary serotonergic neurons is linked to motor output. These neurons may modulate autonomic outflow, but only in relationship to their primary role in motor control. Finally, medullary serotonergic neurons may play a protective role in maintaining glucose homeostasis by disfacilitating the output of the somatomotor system, and hence diminishing muscle energy demands, when peripheral glucose availability is low.

Chemosensitive serotonergic neurons are closely associated with large medullary arteries.

We have previously shown that serotonergic neurons of the medulla are strongly stimulated by an increase in CO(2), suggesting that they are central respiratory chemoreceptors. Here we used confocal imaging and electron microscopy to show that neurons immunoreactive for tryptophan hydroxylase (TpOH) are tightly apposed to large arteries in the rat medulla. We used patch-clamp recordings from brain slices to confirm that neurons with this anatomical specialization are chemosensitive. Serotonergic neurons are ideally situated for sensing arterial blood CO(2), and may help maintain pH homeostasis via wide-ranging effects on brain function. The results reported here support a recent proposal that sudden infant death syndrome (SIDS) results from a developmental abnormality of medullary serotonergic neurons.

Single-unit responses of serotonergic medullary and pontine raphe neurons to environmental cooling in freely moving cats

Brain serotonin has long been implicated in the regulation of body temperature, although its precise role is not completely understood. The present study examined the effects of environmental cooling (4–8°C for 2 or 4 h) on the single-unit activity of serotonergic neurons recorded in the medullary raphe nuclei obscurus and pallidus and in the pontine dorsal raphe nucleus of freely moving cats. These neuronal groups have primarily descending projections to the spinal cord and ascending projections to the forebrain, respectively. Cold exposure induced shivering and piloerection, but no appreciable changes in core temperature. Of the medullary serotonergic cells studied ( n=14), seven were activated and seven were unresponsive to cold exposure. For the responsive cells, the mean increase and peak effect in unit activity relative to baseline were 31% and 46%, respectively. Of the seven cold-responsive cells, the activity of four was monitored when the animals were transferred back to room temperature (23°C). Within 15–30 min, the activity of these cells returned to baseline. In contrast, none of the dorsal raphe nucleus cells studied ( n=14) displayed a significant change in neuronal activity during cold exposure, suggesting that these neurons do not receive afferent input from cold-sensitive cutaneous receptors or participate in thermoregulatory responses evoked by low ambient temperatures. Overall, these results suggest that a subset of medullary serotonergic neurons play a role in physiological mechanisms underlying cold defense (e.g. increases in motor output and/or autonomic outflow). On the other hand, the lack of responsiveness of serotonergic dorsal raphe nucleus neurons to cold exposure does not support a specific role for these cells in thermoregulation.

Do medullary serotonergic neurons tonically modulate nociceptive transmission?

Mason and colleagues have developed an analytical method to reliably identify serotonergic neurons in the rostral ventromedial medulla (RVM), including the raphe magnus, during extracellular recordings in vivo. Results of these studies strongly suggest that the serotonergic neurons do not belong to the major classes of opioid-sensitive neurons phasically activated or inhibited during noxious stimuli. However, their studies have not ruled out any involvement of serotonergic RVM neurons in supraspinal opioid antinociception, and they are at odds with detailed in vitro studies of opioid mechanisms in RVM neurons. It remains possible that tonic activity serotonergic RVM neurons plays an important role in descending opioid modulation of nociception.

Single-unit responses of serotonergic dorsal raphe neurons to specific motor challenges in freely moving cats

Serotonin has been hypothesized to play an important role in the central control of motor function. Consistent with this hypothesis, virtually all serotonergic neurons within the medullary nuclei raphe obscurus and raphe pallidus in cats are activated in response to specific motor challenges. [29]To determine whether the response profile of serotonergic neurons in the midbrain is similar to that observed in the medulla, the single-unit activity of serotonergic dorsal raphe nucleus cells was studied during three specific motor activities: treadmill-induced locomotion, hypercarbia-induced ventilatory response and spontaneous feeding. In contrast to the results obtained for medullary raphe cells, none of the serotonergic dorsal raphe cells studied ( n=26) demonstrated increased firing during treadmill-induced locomotion. A subset of serotonergic dorsal raphe cells (8/36) responded to the hypercarbic ventilatory challenge with increased firing rates that were directly related to the fraction of inspired carbon dioxide, and a non-overlapping subset of cells (6/31) was activated during feeding. All feeding-on cells demonstrated a rapid activation and de-activation coincident with feeding onset and offset, respectively. Although the proportions of serotonergic cells activated by hypercarbia or feeding in the dorsal raphe nucleus were similar to those found in the medullary raphe, there were several major distinctions in the response characteristics for the two cell groups. In contrast to the medullary serotonergic neurons, only a minority of dorsal raphe nucleus serotonergic neurons responded to a motor challenge. Overall, the above results suggest very different roles for the midbrain and medullary serotonergic neurons in response to motor activities.