Intermediate Nucleus Tractus Solitarius Research Articles

Relevance of differential control of sympathorespiratory response magnitudes in clinical assessments.

Relevance of differential control of sympathorespiratory response magnitudes in clinical assessments.

RETRACTED: Sustained Hypoxia Reduces GABAergic Modulation on NTS Neurons Sending Projections to Ventral Medulla of Rats

Peripheral chemoreflex is activated during short-term sustained hypoxia (SH), and the first synapse of these afferents is located in Nucleus Tractus Solitarius(NTS). NTS neurons projecting to the ventral lateral medulla (NTS-VLM) are part of the respiratory pathways of the chemoreflex. SH increases the magnitude of basal respiratory parameters in rats from Wistar-Hannover strain. In this study, we hypothesized that the observed changes in the respiratory pattern in response to SH were due to changes in the GABAergic modulation of the synaptic transmission of NTS-VLM neurons. We used an electrophysiological approach to record the synaptic activity of NTS neurons labeled with a retrograde tracer previously microinjected into VLM of Wistar-Hannover rats submitted to 24 h SH. The data are showing that: (a) the amplitude of evoked inhibitory currents in NTS-VLM neurons of SH rats was reduced and not accompanied by any change in rise-time and decay-time; (b) the 1/CV2 and the number of failures in response to evoked currents were also affected by SH; (c) the frequency of spontaneous inhibitory currents was reduced by SH without changes in amplitude and half-width. These effects of SH were observed in NTS-VLM neurons located in caudal and intermediate NTS, but not in NTS-VLM neurons located in the rostral NTS. We conclude that SH causes a reduction in inhibitory modulation onto NTS-VLM neurons by pre-synaptic mechanisms, which may contribute to the observed changes in the respiratory pattern of Wistar-Hannover rats submitted to SH.

Inhibition of glial glutamate transporter GLT1 in the nucleus tractus solitarii attenuates baroreflex control of sympathetic nerve activity and heart rate

Astrocyte glutamate transporter (GLT‐1) removes neuronally released glutamate from the extracellular space. We examined the effect of inhibiting GLT1 in the nucleus tractus solitarii (NTS) on mean arterial pressure (MAP), renal sympathetic nerve activity (RSNA) and heart rate (HR) in Inactin anesthetized and artificially ventilated Sprague‐Dawley rats. In experiment 1, it was found that dihydrokainate (DHK; inhibitor of GLT1, 5mM, 100nl) injections into the NTS (n=4) decreased MAP (49±12mmHg), RSNA (84±15%) and HR (34±5bpm). Application of kynurenate (KYN; glutamate receptor antagonist, 5mM, 30μl) to the dorsal surface of the brainstem (n=4) blocked responses to NTS injections of DHK with MAP now only decreasing by 14±12mmHg, RSNA by 26±27% and HR by 10±8bpm (p<0.001). The DHK response returned after 30min. In experiment 2, the effect of DHK on arterial baroreflex was examined using i.v. infusions of phenylephrine and nitroprusside. NTS infusions of DHK (5mM, 40nl/min; n=6) reduced baroreflex response range (maximum ‐ minimum) of RSNA by 91±2% and HR by 83±5% (p<0.001). These results indicate that inhibition of GLT1 decreases MAP, RSNA and HR by indirect activation of ionotropic glutamate receptors and that the astrocytic glutamate transporter in the intermediate NTS plays an important role in the maintenance and regulation of cardiovascular function. Supported by HL‐088052

P01.45. Activated neuropathway from nucleus tractus solitarius to rostal ventrolateral medulla during electroacupuncture

Purpose Our previous studies have shown that electroacupuncture (EA) at the Jianshi-Neiguan acupoints (P5-P6, overlying the median nerve) attenuates sympathoexcitatory responses through its influence on neuronal activity in the rostral ventrolateral medulla (rVLM). Nucleus tractus solitarius (NTS) receives inputs from somatic nerve stimulation. However, there is no information on the activation of NTS neurons by EA at P5-P6 acupoints, which subsequently affects the rVLM. Thus, the present study evaluated neuronal activation of NTS in response to EA, with regard to their projections to the rVLM. Methods Seven to ten days after unilateral microinjection of a rodamine-conjugated microsphere retrograde tracer (100 nl) into the rVLM, rats were subjected to EA or served as a sham-operated control. EA was performed for 30 min at P5-P6 acupoints bilaterally. Results Perikarya containing the microsphere tracer were found in the NTS of both groups. Compared to controls (needle placement without electrical stimulation, n=4), c-Fos immunoreactivity and neurons double-labeled with cFos, an immediate early gene and the tracer were more frequently found in the NTS of EA-treated rats (n=5), particularly, in the medial and lateral subdivisions of caudal and intermediate NTS extension. Conclusion These results suggest that EA at P5-P6 acupoints activates NTS neurons. Furthermore, EA-activated NTS neuron can directly project to the rVLM, which is known to participate in EA-modulation of sympathetic activity.

Astrocytes in NTS after exposure to chronic intermittent hypoxia

Chronic intermittent hypoxia (CIH) is a model of the arterial hypoxemia that accompanies sleep apnea and increases resting arterial pressure. Exposures to hypoxia activate astrocytes in the brain. Astrocytes are a primary source of brain angiotensinogen, the precursor to angiotensin II. We tested the hypothesis that chronic intermittent hypoxia increases astrocytes in the nucleus tractus solitarii (NTS). Sprague‐Dawley rats (13 ± 0.3 weeks old) were exposed to CIH (9% oxygen for 3min every 10 min, 8 h per day) for 7 days (n = 5) while control rats (n = 4) were maintained in normoxia. After the CIH protocol, brain stem tissue containing NTS was obtained. Protein levels of the astrocyte marker glial fibrillary acidic protein (GFAP) in the NTS were assessed by Western blot analysis. Relative protein levels of GFAP to actin in the caudal NTS did not differ between CIH and control groups (3.3 ± 1.7 vs. 4.0 ± 1.7, CIH vs. control, P=0.59). The GFAP levels tended to be lower in CIH than in the control group in intermediate and rostral NTS (intermediate: 2.3 ± 1.1 vs. 3.7 ± 0.6, P=0.11; rostral: 1.5 ± 0.9 vs. 2.7 ± 1.0, P=0.052). These results did not support the idea that hypertension induced by CIH is associated with an activation of astrocytes in the NTS. CIH might decrease astrocytes in the NTS as opposed to our hypothesis. Supported by HL‐088052

Decrease in the expression of N‐methyl‐D‐aspartate receptors in the nucleus tractus solitarii induces antinociception and increases blood pressure

N-methyl-D-aspartate receptors (NMDAR) have a role in cardiovascular control at the nucleus tractus solitarii (NTS), eliciting increases or decreases in blood pressure (BP), depending on the area injected with the agonists. In spite of the association between cardiovascular control and pain modulation, the effects of manipulating NMDAR in pain responses have never been evaluated. In this study, we decreased the expression of NMDAR in the NTS using gene transfer to target receptor subunits and evaluate long-term effects. Seven days after the injection of lentiviral vectors containing the NR1a subunit cDNA of NMDAR, in antisense orientation, into the intermediate NTS of Wistar rats, BP was measured, and the formalin test of nociception was performed. The antisense vector induced a decrease of NR1 expression in the NTS and elicited BP rises and hypoalgesia. Antisense vectors inhibited formalin-evoked c-Fos expression in the spinal cord, indicating decreased nociceptive activity of spinal neurons. Using a time-course approach, we verified that the onset of both the increases in BP and the hypoalgesia was at 4 days after vector injection into the NTS. The injection of NMDA into the NTS reversed the effects of antisense vectors in pain behavioral responses and spinal neuronal activation and decreased BP and heart rate. The present study shows that the NR1 subunit of the NMDAR at the NTS is critical in the regulation of tonic cardiovascular and nociceptive control and shows an involvement of the nucleus in the modulation of sustained pain.

Inhibition of brown adipose tissue thermogenesis by neurons in the ventrolateral medulla and in the nucleus tractus solitarius

Neurons in the ventrolateral medulla (VLM) and in the nucleus tractus solitarius (NTS) play important roles in the regulation of cardiovascular and other autonomic functions. In the present study, we demonstrate an inhibition of brown adipose tissue (BAT) thermogenesis evoked by activation of neurons in the VLM, as well as by neurons in the intermediate NTS, of chloralose/urethane-anesthetized, artificially ventilated rats. Activation of neurons in either rostral VLM or caudal VLM with N-methyl-d-aspartate (12 nmol) reversed the cold-evoked increase in BAT sympathetic nerve activity (SNA), BAT temperature, and end-expired CO(2). Disinhibition of neurons in either VLM or NTS with the GABA(A) receptor antagonist, bicuculline (30 pmol), reversed the increases in BAT SNA, BAT temperature, and end-expired CO(2) that were elicited 1) by cold defense; 2) during the febrile model of nanoinjection of prostaglandin E(2) into the medial preoptic area; 3) by activation of neurons in the dorsomedial hypothalamus or in the rostral raphe pallidus (rRPa); or 4) by the micro-opioid receptor agonist fentanyl. Combined, but not separate, inhibitions of neurons in the VLM and in the NTS, with the GABA(A) receptor agonist, muscimol (120 pmol/site), produced increases in BAT SNA, BAT temperature, and expired CO(2), which were reversed by nanoinjection of glycine (30 nmol) into the rRPa. These findings suggest that VLM and NTS contain neurons whose activation inhibits BAT thermogenesis, that these neurons receive GABAergic inputs that are active under these experimental conditions, and that neurons in both sites contribute to the tonic inhibition of sympathetic premotor neuronal activity in the rRPa that maintains a low level of BAT thermogenesis in normothermic conditions.

Glutamatergic Antagonism in the NTS Decreases Post-Inspiratory Drive and Changes Phrenic and Sympathetic Coupling During Chemoreflex Activation

For a better understanding of the processing at the nucleus tractus solitarius (NTS) level of the autonomic and respiratory responses to peripheral chemoreceptor activation, herein we evaluated the role of glutamatergic neurotransmission in the intermediate (iNTS) and caudal NTS (cNTS) on baseline respiratory parameters and on chemoreflex-evoked responses using the in situ working heart-brain stem preparation (WHBP). The activities of phrenic (PND), cervical vagus (cVNA), and thoracic sympathetic (tSNA) nerves were recorded before and after bilateral microinjections of kynurenic acid (Kyn, 5 nmol/20 nl) into iNTS, cNTS, or both simultaneously. In WHBP, baseline sympathetic discharge markedly correlated with phrenic bursts (inspiration). However, most of sympathoexcitation elicited by chemoreflex activation occurred during expiration. Kyn microinjected into iNTS or into cNTS decreased the postinspiratory component of cVNA and increased the duration and frequency of PND. Kyn into iNTS produced no changes in sympathoexcitatory and tachypneic responses to peripheral chemoreflex activation, whereas into cNTS, a reduction of the sympathoexcitation, but not of the tachypnea, was observed. The pattern of phrenic and sympathetic coupling during the chemoreflex activation was an inspiratory-related rather than an expiratory-related sympathoexcitation. Kyn simultaneously into iNTS and cNTS produced a greater decrease in postinspiratory component of cVNA and increase in frequency and duration of PND and abolished the respiratory and autonomic responses to chemoreflex activation. The data show that glutamatergic neurotransmission in the iNTS and cNTS plays a tonic role on the baseline respiratory rhythm, contributes to the postinspiratory activity, and is essential to expiratory-related sympathoexcitation observed during chemoreflex activation.

Fos expression in the NTS in response to peripheral chemoreflex activation in awake rats

Chemoreflex afferent fibers terminate in the nucleus tractus solitarii (NTS), but the specific location of the NTS neurons excited by peripheral chemoreflex activation remains to be characterized. Here, the topographic distribution of chemoreflex sensitive cells at the commissural NTS was evaluated. To reach this goal, Fos-immunoreactive neurons (Fos-ir) were accounted in rostro-caudal levels of the intermediate and caudal commissural NTS, after intermittent chemoreflex activation with intravenous injection of potassium cyanide [KCN (80 μg/kg) or saline (0.9%, vehicle), one injection every 3 min during 30 min]. In response to intermittent intravenous injections of KCN, a significant increase in the number of Fos-ir neurons was observed specifically in the lateral intermediate commissural NTS [ LINTS (82 ± 9 vs. 174 ± 16, cell number mean per section)] and lateral caudal commissural NTS [ LCNTS (71 ± 9 vs. 199 ± 18, cell number mean per section)]. To evaluate the influence of baroreceptor-mediated inputs following the increase in blood pressure during intermittent chemoreflex activation, we performed an intermittent activation of the arterial baroreflex by intravenous injection of phenylephrine [1.5 μg/kg iv (one injection every 3 min during 30 min)]. This procedure induced no change in Fos-ir in LINTS (64 ± 6 vs. 62 ± 12, cell number mean per section) or LCNTS (56 ± 15 vs. 77 ± 12, cell number mean per section). These data support the involvement of the commissural NTS in the processing of peripheral chemoreflex, and provide a detailed characterization of the topographical distribution of activated neurons within this brain region.

Cardiovascular responses to hydrogen peroxide into the nucleus tractus solitarius

The nucleus tractus solitarius (NTS), a major hindbrain area involved in cardiovascular regulation, receives primary afferent fibers from peripheral baroreceptors and chemoreceptors. Hydrogen peroxide (H(2)O(2)) is a relatively stable and diffusible reactive oxygen species (ROS), which acting centrally, may affect neural mechanisms. In the present study, we investigated effects of H(2)O(2) alone or combined with the glutamatergic antagonist kynurenate into the NTS on mean arterial pressure (MAP) and heart rate (HR). Conscious or anesthetized (urethane and alpha-chloralose) male Holtzman rats (280-320 g) were used. Injections of H(2)O(2) (125 to 1500 pmol/40 nl) into the intermediate NTS of anesthetized rats evoked dose-dependent and transient hypotension (-18 +/- 3 to -55 +/- 11 mmHg) and bradycardia (-16 +/- 5 to -116 +/- 40 bpm). Injection of the catalase inhibitor 3-amino-1,2,4-triazole (100 nmol/40 nl) into the NTS also produced hypotension and bradycardia. Previous injection of the ionotropic L-glutamate receptor antagonist kynurenate (7 nmol/40 nl) attenuated by 48% the bradycardic response, without changing the hypotension evoked by H(2)O(2) (500 pmol/40 nl) in anesthetized rats. The antioxidant L-ascorbate (600 pmol/80 nl) injected into the NTS attenuated the bradycardic (42%) and hypotensive (67%) responses to H(2)O(2) (500 pmol/40 nl) into the NTS. In conscious rats, injection of H(2)O(2) (50 nmol/100 nl) into the NTS also evoked intense bradycardia (-207 +/- 8 bpm) and hypotension (-54 +/- 6 mmHg) that were abolished by prior injection of kynurenate (7 nmol/100 nl). The results show that H(2)O(2) into the NTS induces hypotension and bradycardia probably due to activation of glutamatergic mechanisms.

Hypotension and short‐term anaesthesia induce ERK1/2 phosphorylation in autonomic nuclei of the brainstem

The aims of this study were: first, to investigate the effects of anaesthesia on phosphorylated extracellular signal-regulated kinase (p-ERK)1/2-immunoreactivity (-ir) in the brainstem; second, to choose the best anaesthetic for p-ERK1/2 studies; and third, to determine the effect of short-term hypotension on p-ERK1/2-ir in the brainstem. Rats were anaesthetized with halothane, sodium pentobarbital or 100% CO2 narcosis, or were cervically dislocated and within 5 min perfused and the brains processed immunohistochemically for pERK1/2-ir. p-ERK1/2-ir was primarily observed in regions associated with cardiovascular and/or respiratory control. Several regions consistently showed dense p-ERK1/2 labelling, including a restricted region of the ventrolateral medulla (VLM). In contrast, other regions showed differential labelling depending on the mode of death. Cervical dislocation showed the least VLM labelling, limited to a discrete area approximately 0.6-1.4 mm caudal to the facial nucleus. Anaesthetics induced labelling throughout the VLM, with halothane inducing the most. Many p-ERK1/2-ir VLM neurons were catecholaminergic following halothane or sodium pentobarbital anaesthesia, but no double labelling was seen following cervical dislocation. Of the anaesthetics, sodium pentobarbital induced the least labelling and was used subsequently. Intravenous hydralazine was used to induce a 20-min period of hypotension, whereas arterial pressure did not change in vehicle-treated animals. Hydralazine evoked more pERK-ir neurons in specific regions, including the VLM, nucleus tractus solitarius (NTS), parabrachial nuclei, Kolliker-Fuse nucleus and locus coeruleus. Approximately twice as many p-ERK1/2-positive neurons were seen in the intermediate NTS and rostral VLM following hydralazine compared with the vehicle. In conclusion, p-ERK1/2-ir identifies neurons in central autonomic regions, and their number and distribution are markedly affected by anaesthetics, and are increased in some regions by short-term hypotension.

AUTONOMIC and RESPIRATORY RESPONSES TO MICROINJECTION OF ATP INTO THE INTERMEDIATE OR CAUDAL NUCLEUS TRACTUS SOLITARIUS IN THE WORKING HEART–BRAINSTEM PREPARATION OF THE RAT

1. Activation of peripheral chemoreceptors with KCN in the working heart-brainstem preparation from young male Wistar rats (70-90 g) increases phrenic (PNA; +105 +/- 18%) and thoracic (tSNA; +44 +/- 6%) sympathetic nerve activity compared with baseline and reduces heart rate (HR; from 377 +/- 27 to 83 +/- 6 b.p.m.). 2. Microinjections of increasing doses of ATP (1, 5, 25, 100 and 500 mmol/L; n = 7) into the intermediate nucleus tractus solitarius (NTS) produced a dose-dependent reduction in PNA (from -6 +/- 3 to -82 +/- 1%) and in HR (from -12 +/- 4 to -179 +/- 47 b.p.m.). Microinjections of ATP into the intermediate NTS also produced a reduction in tSNA (from -3 +/- 3 to -26 +/- 5%), which was not dose dependent. 3. Microinjections of ATP into the caudal NTS (n = 5) produced a dose-dependent increase in PNA (from 0.2 +/- 3 to 115 +/- 27%) and minor changes in HR and tSNA, which were not dose dependent. 4. The data show that microinjection of ATP into distinct subregions of the NTS produces different respiratory and autonomic responses and suggest that ATP in the caudal NTS is involved in the respiratory but not in the sympathoexcitatory component of the chemoreflex.

Hemodynamic and respiratory responses to microinjection of ATP into the intermediate and caudal NTS of awake rats

The nucleus tractus solitarii (NTS) is the site of integration of the peripheral chemoreceptor afferents in the brainstem. Previous studies from our laboratory have shown that microinjection of ATP into the intermediate NTS produced increases in arterial pressure and bradycardia. In the present study, we evaluated the hemodynamic and respiratory responses to microinjection of ATP into the intermediate and caudal commissural NTS. In the same group of rats the responses were compared with cardiorespiratory responses to chemoreflex activation (KCN, i.v.). The data show that microinjection of ATP into the intermediate NTS produced pressor and bradycardic responses similar to those observed in response to chemoreflex activation but apnoea instead of tachypnoea. Microinjection of ATP into caudal commissural NTS produced increase in arterial pressure and tachypnoea similar to the chemoreflex but a minor bradycardia. The data show that microinjection of ATP into different sub-regions of the NTS produces a diverse pattern of hemodynamic and respiratory responses and suggest the involvement of this purine in the neurotransmission of the cardiovascular reflex in the NTS.

Antagonism of glutamate receptors in the intermediate and caudal NTS of awake rats produced no changes in the hypertensive response to chemoreflex activation

The role of excitatory amino acid (EAA) receptors in the neurotransmission of the sympathoexcitatory component of the chemoreflex (pressor response) in the intermediate and caudal aspects of the commissural nucleus tractus solitarii (NTS) of awake rats was evaluated. Microinjection of kynurenic acid, a non-selective antagonist of EAA receptors, into the intermediate and caudal commissural NTS produced a large increase in the baseline mean arterial pressure (MAP), which may reduce the magnitude of the pressor response to chemoreflex activation. To avoid this problem sodium nitroprusside (SNP) was infused (i.v.) after microinjections of kynurenic acid (2 nmol/50 nl) into the NTS, in order to normalize the MAP and then the chemoreflex was activated and the magnitude of the pressor response evaluated. Microinjection of kynurenic acid into the intermediate (bilaterally) and caudal (midline) commissural NTS ( n=6) produced a significant increase in baseline MAP (103±5 vs. 137±6 mm Hg) normalized by SNP infusion (107±4 mm Hg) and under this experimental condition the pressor response to chemoreflex activation was not statistically different in relation to the control (37±7 vs. 44±6 mm Hg). Bilateral microinjections of kynurenic acid into the caudal NTS ( n=8) also produced a significant increase in baseline MAP (109±4 vs. 145±6 mm Hg) normalized by SNP infusion (109±6 mm Hg). After normalization of MAP, the pressor response to chemoreflex activation at 3 (34±6 mm Hg) and 10 min (37±6 mm Hg) was also not different in relation to the control (46±5 mm Hg). These data indicate that the antagonism of EAA receptors simultaneously in the intermediate (bilateral) and caudal (midline) commissural NTS or only in the caudal commissural NTS (bilateral) of awake rats had no effect on the hypertensive response to chemoreflex activation. We suggest that neurotransmitter other than l-glutamate may take part in the neurotransmission of the sympathoexcitatory component of the chemoreflex at the NTS level.

Glutamatergic neural transmission in the nucleus tractus solitarius: N-methyl-D-aspartate receptors.

1. The nucleus tractus solitarius (NTS) is the first central site where the reflex control of autonomic, including baroreceptor, reflex function is coordinated. Autonomic signals are transmitted from the first-order visceral afferent fibres to second-order NTS neurons by L-glutamate. It is well established that activation of the alpha-amino-3-hydroxy-5-methyl-4-isoxazole proprionic acid (AMPA) receptors, which mediate the fast component of L-glutamate signalling, is required for generating changes in membrane potentials of the second-order NTS neurons. The contribution of the slower-developing, longer-lasting N-methyl-D-aspartate (NMDA) receptor-mediated component of glutamate signalling to synaptic transmission at these synapses is less well understood. 2. The aim of this work is to highlight evidence that functional NMDA receptors exist on second-order NTS neurons in autonomic, including baroreceptor, afferent pathways by determining whether NMDA receptors can be activated by: (i) exogenous application of NMDA; and (ii) endogenous release of L-glutamate from autonomic afferent fibres. Studies were performed on second-order neurons in transverse and horizontal brainstem slices containing the intermediate NTS and the tractus solitarius. Second-order NTS neurons were identified by electrophysiological criteria or by attached fluorescent-labelled aortic depressor nerve (ADN) boutons. 3. N-Methyl-D-aspartate (50 nmol(-2) micromol) dose-dependently evoked excitatory post-synaptic currents in second-order NTS neurons (P = 0.004; n = 4). The NMDA receptor-mediated currents were also synaptically evoked by low-frequency stimulation of the autonomic afferent fibres in the tractus solitarius. The NMDA receptor-mediated currents were blocked by the NMDA receptor antagonist AP5 (n = 7; P = 0.027). 4. The findings suggest that functional NMDA receptors exist on second-order NTS neurons. While the NMDA receptor- mediated currents may not be required for signal transmission when the second-order neurons are at resting membrane potential, their activation may help to modulate autonomic signal transmission in the NTS under conditions in which the membrane is depolarized by high frequency or convergent inputs.

Central neurocircuitry associated with emesis

Ingestion of toxin, traumatic events, adverse drug reactions, and motion can all result in nausea and emesis. In addition, cyclic vomiting syndrome is quite prevalent in the pediatric population. Coordination of the various autonomic changes associated with emesis occurs at the level of the medulla oblongata of the hindbrain. Chemosensitive receptors detect emetic agents in the blood and relay this information by means of neurons in the area postrema to the adjacent nucleus tractus solitarius (NTS). Abdominal vagal afferents that detect intestinal luminal contents and gastric tone also terminate in the NTS (gelatinosus, commissural, and medial subnuclei). The NTS is viscerotopically organized into subnuclei that subserve diverse functions related to swallowing (subnucleus centralis), gastric sensation (subnucleus gelatinosus), laryngeal and pharyngeal sensation (intermediate and interstitial NTS), baroreceptor function (medial NTS), and respiration (ventrolateral NTS). Neurons from the NTS project to a central pattern generator (CPG), which coordinates the sequence of behaviors during emesis, as well as directly to diverse populations of neurons in the ventral medulla and hypothalamus. Thus, it is critical to realize that there is not an isolated “vomiting center,” but rather groups of loosely organized neurons throughout the medulla that may be activated in sequence by a CPG. The newer antiemetic agents appear to block receptors in the peripheral endings of vagal afferents to reduce “perception” of emetic stimuli and/or act in the dorsal vagal complex. A primary site of action of 5-HT 3-receptor antagonists is by means of the vagal afferents. Neurokinin-1 receptor (NK 1R) antagonists are antiemetics, because they act at a site in the dorsal vagal complex. Part of their effectiveness may be the result of inhibition of the NK 1R on vagal motor neurons to prevent fundic relaxation, which is a prodromal event essential for emesis. Delta 9-tetrahydrocannabinol (Δ 9-THC), the major psychoactive component of marijuana, can be therapeutically useful as an antiemetic. The site of action of Δ 9-THC is on cannabinoid CB1 receptors in the dorsal vagal complex. However, it decreases fundic tone and antral motility. It is not easy to predict the potential antiemetic effects of drugs that alter motility. Although antiemetic drugs are available for management of acute chemotherapeutic-induced emesis, few treatments are effective for delayed emesis or cyclic vomiting syndrome.

Mu-opioid receptors are present in vagal afferents and their dendritic targets in the medial nucleus tractus solitarius.

Ligands of the mu-opiate receptor (MOR) are known to influence many functions that involve vagal afferent input to the nucleus tractus solitarius (NTS), including cardiopulmonary responses, gastrointestinal activity, and cortical arousal. The current study sought to determine whether a cellular substrate exists for direct modulation of vagal afferents and/or their neuronal targets in the NTS by ligands of the MOR. Anterograde tracing of vagal afferents arising from the nodose ganglion was achieved with biotinylated dextran amine (BDA), and the MOR was detected by using antipeptide MOR antiserum. The medial subdivision of the intermediate NTS was examined by electron microscopy for the presence of peroxidase-labeled, BDA-containing vagal afferents and immunogold MOR labeling. MOR was present in both presynaptic axon terminals and at postsynaptic sites, primarily dendrites. In dendrites, MOR immunogold particles usually were located along extrasynaptic portions of the plasma membrane. Of 173 observed BDA-labeled vagal afferent axon terminals, 33% contained immunogold labeling for MOR within the axon terminal. Many of these BDA-labeled terminals formed asymmetric, excitatory-type synapses with dendrites, some of which contained MOR immunogold labeling. MORs were present in 19% of the dendrites contacted by BDA-labeled terminals but were present rarely in both the vagal afferent and its dendritic target. Together, these results suggest that MOR ligands modulate either the presynaptic release from or the postsynaptic responses to largely separate populations of vagal afferents in the intermediate NTS. These results provide a cellular substrate for direct actions of MOR ligands on primary visceral afferents and their second-order neuronal targets in NTS.

The gain of the baroreflex bradycardia is reduced by microinjection of NMDA receptor antagonists into the nucleus tractus solitarii of awake rats

The baroreflex activation with phenylephrine infusion produces a bradycardic response. In the present study, the role of NMDA receptors in the nucleus tractus solitarii (NTS) in the processing of the parasympathetic component of the baroreflex was evaluated using acid phosphonivaleric (AP-5), a selective NMDA receptor antagonist. Baroreflex activation was performed before and after bilateral microinjection of AP-5 into the intermediate commissural NTS (0.5 mm lateral to the midline). Microinjection of the vehicle (saline, 0.9%) or a dose of 2 nmol/50 nl of AP-5 into the NTS produced no effect on the gain of the baroreflex while a dose of 10 nmol/50 nl of AP-5 produced a significant reduction in the gain of the baroreflex 2 min after microinjection [−1.43±0.22 vs. −0.43±0.03 bpm/mmHg, ( n=6)], with a return to control levels 10 min after the microinjections. The dose of 10 nmol/50 nl was selective for NMDA receptors considering that the cardiovascular responses to microinjection of AMPA (0.05 pmol/50 nl), a non-NMDA receptor agonist, were not affected by this dose of AP-5 and the responses to microinjection of NMDA (2 nmol/50 nl) were blocked. The data show that the bradycardic response to baroreflex activation was blocked by AP-5 microinjected into the NTS, indicating that the neurotransmission of the parasympathetic component of the baroreflex is mediated by NMDA receptors in the NTS.

The distribution of neuronal nitric oxide synthase in the nucleus tractus solitarii of the squirrel monkey

The distribution of neuronal nitric oxide synthase (nNOS) containing neurons and fibers in subnuclei of the nucleus tractus solitarii (NTS) in the squirrel monkey, Saimuri sciureus, was investigated by nNOS immunohistochemistry and nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase histochemistry. Generally, the staining pattern of nNOS and NADPH-diaphorase in the NTS was similar. A high density of neurons and fibers exhibiting both nNOS immunoreactivity and NADPH-diaphorase reactivity was present in the central, medial, intermediate, and dorsolateral subnuclei of the NTS. A moderate density of neurons and fibers that stained for both nNOS and NADPH-diaphorase was noted in the interstitial and ventromedial subnuclei. The gelatinosus and commissural subnuclei contained a low density of neurons and fibers exhibiting nNOS immunoreactivity and NADPH-diaphorase staining. The dorsal motor nucleus of vagus contained a high density of nNOS immunopositive and NADPH-diaphorase containing neurons and fibers at the rostral level, but contained a moderate density of positive fibers and very few positive neurons at the intermediate, subpostremal and commissural NTS levels. Incongruence was noted, however, between nNOS immunostaining and NADPH-diaphorase staining in blood vessels in the brainstem. Capillaries and small vessels exhibited strong staining for NADPH-diaphorase but no nNOS immunoreactivity. In summary, this work substantiates the presence of nNOS in subnuclei of the monkey NTS and is consistent with a role for NO · in neurotransmission in primate NTS.

Modulation of the carotid baroreceptor reflex by substance P in the nucleus tractus solitarius

Previous studies have shown that administration of substance P (SP) into the nucleus tractus solitarius (NTS) can evoke a depressor response similar to that produced by activation of the arterial baroreceptors. In addition, some studies have suggested that SP increases the reflex responses to activation of baroreceptor input. The present study was performed to determine the effects of SP on the carotid sinus baroreceptor reflex at the level of the NTS by examining the effects of both exogenous SP microinjected into different rostrocaudal locations in the NTS and blockade of the effects of endogenous SP, through the microinjection of a substance P antagonist (SPa; [ d-Pro, d-Trp]-substance P). Changes in pressure in an isolated carotid sinus in anesthetized dogs were used to evoke baroreflex changes in arterial blood pressure (BP) before and after microinjection of SP (0.5 μM) or SPa (10 μM) into barosensitive regions of the NTS. Microinjection of SP or its antagonist did not alter baseline, resting BP but did produce significant changes in baroreflex sensitivity. Microinjection of SP into different rostrocaudal regions of the NTS produced different responses, with rostral and caudal NTS microinjections producing significant increases in sensitivity. No effects on baroreflex sensitivity were obtained in response to SP microinjections into the intermediate NTS. Unlike SP, microinjection of the SPa significantly decreased baroreflex sensitivity at all rostrocaudal levels of the NTS. These data demonstrated that SP has the capability to modulate the carotid baroreflex at the level of the NTS and support a physiological role for endogenously released SP.