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Cranial nerves and ganglia are altered after in vitro treatment of mouse embryos with valproic acid (VPA) and 4-en-VPA.

Prenatal valproic acid (VPA) exposure results in neural tube defects and in the fetal valproate syndrome (FVS), associated with developmental delay. In the present study we investigate the alterations induced by VPA and one of its metabolite, 4-en-VPA, on specific neural structures: branchial nerves and ganglia. This study was performed on 8-9 pairs of somites mouse embryos exposed in vitro for 24 h to 0.75 mM of VPA or 1 mM of 4-en-VPA. After an additional culture period of 20 h without drug, the embryos were processed for whole mount immunostaining using the monoclonal antibody 2H3, directed against the 155 kDa neurofilament protein. This technique makes it possible to visualise the branchial nerves/ganglia. VPA and 4-en-VPA induced a delay in the development of the trigeminal (V), glossopharyngeal (IX) and vagus (X) nerves/ganglia. The development of the facial (VII) nerve was delayed to a lesser extend. These treatments also induced defects in the four ganglia. The main abnormalities were a reduced dorsal component of ganglion V, the absence of the dorsal root of ganglion IX, a disorganised dorsal part of ganglion X and diffuse ventral fibres in nerves VII-VIII. In addition, scattered fibres were observed around and between ganglia. In conclusion, VPA and 4-en-VPA deeply altered the differentiation of branchial nerves/ganglia. The dorsal part of the ganglia, arising from the rhombencephalic neural crest, was particularly sensitive. The disorganisation of fibres could possibly be explained by alteration of the extracellular matrix.

Animals↗

Direct contacts between Auerbach's ganglia and smooth muscle cells in the small intestine of the rat.

The present electron microscopic study demonstrated direct contacts between Auerbach's ganglia and longitudinal smooth muscle cells in the rat small intestine. The muscle cells were often observed to extend small, foot-like processes to the Auerbach's ganglia. These processes were in contact with glial cells in the ganglia without an intervening basal lamina, or were in contact with intraganglionic axon varicosities containing many synaptic vesicles. The processes in contact with glial cells may anchor the muscle cells to the ganglia during muscle contraction and those in contact with axon varicosities may function as synaptic sites between ganglion and longitudinal muscle cells.

Animals↗

Isolation of a polypeptide from the venom of Bungarus multicinctus that binds to ganglia and blocks the ganglionic transmission in mammals.

A 15,000 dalton polypeptide purified from Bungarus multicinctus venom (which normally copurifies with alpha-bungarotoxin) was characterized biochemically and its biological effects were studied. This polypeptide, P15, had an aminoacid composition and molecular weight different from those of both alpha- and beta-bungarotoxin. It inhibited the ganglionic transmission in the guinea-pig hypogastric nerve-vas deferens preparation and did not block, even at very high concentrations, the neuromuscular transmission in the rat phrenic nerve-diaphragm preparation. In the same preparations alpha-bungarotoxin was unable to block the response at the ganglionic synapse while it was fully active in blocking the neuromuscular transmission. However, a pretreatment of the vas deferens preparation with alpha-bungarotoxin prevented the inhibitory effect of P15. 125I-Labeled P15 showed a specific and saturable binding to rat superior cervical ganglia homogenate and to a Torpedo postsynaptic membrane fraction. The binding of P15 to ganglia was inhibited by curare. The binding was Ca2+ dependent. The density of binding sites was of 300 fmol/mg of protein in the ganglion and 500 fmol/mg of protein in Torpedo membranes. The amount of P15-binding sites in ganglia was not modified by denervation, indicating that P15 binds to postsynaptic receptors. The binding of 125I-labeled P15, both in ganglia and Torpedo membranes, was inhibited by alpha-bungarotoxin. P15 had a Ca2+-dependent phospholipase A2 activity. Lowering Ca2+ concentration in incubation media affected the phospholipase A2 activity more than binding properties and inhibition of phospholipase activity with p-bromophenacyl bromide did not affect the activity of P15 on vas deferens preparation, suggesting that the phospholipase activity is not necessary for the activity of P15 on nicotinic receptors. Our results suggest that P15 toxin may be a specific and valuable probe for studying the ganglionic nicotinic receptor.

Animals↗

Nitric oxide synthase-immunoreactive vagal afferent fibers in rat superior cervical ganglia.

Chronic (5-14 days) preganglionic denervation of the rat superior cervical ganglia by sectioning the cervical sympathetic trunk resulted in a time-related partial or complete loss of nitric oxide synthase (isoform I)-immunoreactive fibers and terminals surrounding many sympathetic ganglionic neurons. Unexpectedly, denervation unmasked many varicose nitric oxide synthase-immunoreactive fibers, some of which could be traced the entire length of the superior cervical ganglia. Injection of the retrograde tracer Fluorogold into the superior cervical ganglia labeled a population of nodose ganglion cells and of dorsal root ganglion cells from C8 to T3 segments. When the same sections were processed for nitric oxide synthase-immunoreactivity, 40% of the Fluorogold-containing nodose ganglion cells also expressed nitric oxide synthase-immunoreactivity, whereas colocalization was observed in only a few dorsal root ganglion cells. Similarly, injection of Fluorogold into denervated superior cervical ganglia labeled a population of nodose ganglion cells. Sectioning of all nerve trunks associated with the superior cervical ganglion prior to injection of Fluorogold, except the cervical sympathetic trunk, resulted in no detectable labeling of Fluorogold in the ipsilateral nodose ganglion cells. These results indicate that a population of rat nodose ganglion cells contain nitric oxide synthase and that some of these neurons project their axons through the superior cervical ganglion and terminate in the peripheral target tissues. The possibility that nitric oxide synthase-immunoreactive vagal afferent fibers may participate in nociception is considered.

Afferent Pathways↗

Reduced CCK-induced Fos expression in the hindbrain, nodose ganglia, and enteric neurons of rats lacking CCK-1 receptors.

Many of the actions of cholecystokinin (CCK) are mediated by CCK-1 receptors, expressed by enteric and vagal afferent neurons. Otsuka Long-Evans Tokushima Fatty rats (OLETF) do not express CCK-1 receptors, and do not exhibit the vagally mediated responses to CCK. To determine whether the OLETF rat's failure to respond to CCK is correlated with failure of CCK to activate enteric and vagal neurons, we quantified neuronal Fos immunoreactivity in the dorsal vagal complex of the hindbrain, the nodose ganglia, and the ganglia of the myenteric and submucosal plexuses of the duodenum following intraperitoneal injection of CCK-8 (20 microg/kg). Compared to vehicle injection, CCK administration resulted in significant increases in the number of Fos-immunopositive neurons in the nucleus of the solitary tract, area postrema, and dorsal vagal motor nucleus of control, LETO rats. In OLETF rats, however, CCK did not increase numbers of Fos-immunoreactive neurons in any of these brain structures. CCK also induced significantly larger numbers of Fos-immunoreactive neuronal nuclei in the nodose ganglia of LETO rats, but not in the nodose ganglia of OLETF rats. Finally, LETO, but not OLETF rats exhibited striking increases in the number of Fos-immunoreactive nuclei of myenteric and submucosal neurons, following CCK injection. Absence of CCK-induced Fos expression in OLETF rats is consistent with attenuation of ingestive and gastrointestinal responses to CCK in the CCK-1 receptor deficient rats. These results also suggest that CCK-induced Fos expression in enteric and vagal sensory neurons of rats can be accounted for entirely by activation of CCK-1 receptors and is not due to occupation of CCK-2 (gastrin) receptors, which also are expressed in the intestine and by some vagal afferent neurons.

Animals↗

Changes in transient receptor potential cation channel superfamily V (TRPV) mRNA expression in the mouse inner ear ganglia after kanamycin challenge.

The transient receptor potential cation channel subfamily V (TRPV) is a non-specific cation ion channel receptor family that is gated by heat, protons, low extracellular osmolarity and arachidonic acid derivatives. Since some of these endogenous agonists of TRPV receptors are reactive oxygen intermediates produced by lipoxygenases, it has been hypothesized that some members of the TRPV family may respond to challenges by reactive oxygen species. This study used real-time PCR to quantitatively track changes in TRPV1-4 mRNA expression in the spiral, vestibular, and trigeminal ganglia and the kidney from kanamycin (KM)-treated mice. TRPV1, TRPV2, TRPV3 and TRPV4 mRNAs were expressed in spiral and vestibular ganglia, and TRPV2 and TRPV1 mRNAs were most predominant in control mice. After KM (700 mg/kg s.c. b.i.d., 14 days), TRPV1 mRNA and protein expression were significantly up-regulated both in the spiral and vestibular ganglia, but expression was unaffected in the trigeminal ganglion and kidney. Real-time PCR also demonstrated a significant down-regulation in TRPV4 mRNA expression in the inner ear ganglia and kidney after KM treatment. All these mRNA and protein expression changes were eliminated by simultaneous administration of dihydroxybenzoate (300 mg/kg s.c. b.i.d., 14 days), an anti-oxidant that blocks KM ototoxicity. It is proposed that up-regulated TRPV1 expression during KM exposure may promote ganglion cell survival by contributing to neuronal depolarization, with KM-induced tinnitus and dizziness as consequences.

Animals↗

Third pathway in the cortico-basal ganglia loop: Neurokinin B-producing striatal neurons modulate cortical activity via striato-innominato-cortical projection.

In the cortico-basal ganglia loop, striatal regions serve as 'entrances' to the basal ganglia, receiving massive inputs from the cerebral cortex and sending 'direct' and 'indirect' pathways to the output nuclei of the basal ganglia. However, we have recently identified a new striatofugal subgroup which produces neurokinin B (NKB). Although NKB-producing neurons constitute a minority of striatal neurons, this subgroup is distinguished by the unique distribution and chemical characteristics. NKB-producing striatal neurons are distributed in association with mu-opioid receptor localization, and rarely express DARPP32, which is produced by the major striatofugal neurons and coupled with dopaminergic signaling. Further interestingly NKB-producing striatal neurons send axons to basal forebrain regions, but not to the main target regions of striatal outflow, pallidal or mesencephalic regions. In the basal forebrain, some GABAergic inhibitory neurons express NK3 receptor, selective receptor for NKB, and directly send axons to the cerebral cortex. The NK3-expressing neurons show different electrical properties from cholinergic basal forebrain neurons, and display facilitatory responses to stimulation of NK3 receptor. These findings strongly suggest that NKB-producing striatal neurons and NK3-expressing basal forebrain neurons constitute a third pathway which bypasses the common output nuclei of the basal ganglia, and more directly control or modulate cortical activity.

Animals↗

Remodelling of connections in pelvic ganglia after hypogastric nerve crush.

Pelvic ganglia innervate the urogenital organs and contain both sympathetic and parasympathetic neurons. Previous studies have shown that within days of cutting either the lumbar or sacral preganglionic axons that innervate pelvic ganglia, many axon collaterals grow and appear to form specific connections with denervated pelvic neurons. Here we have examined the longer term consequences of partial deafferentation by studying pelvic ganglia up to 7 weeks after hypogastric nerve (HGN) crush, a lesion which also allows faster regeneration of spinal axons. Noradrenergic neurons were denervated by HGN crush, as demonstrated by loss of varicosities immunostained for the synaptic proteins, synaptophysin and synapsin. A week after HGN crush, axon collaterals grew from parasympathetic pelvic ganglion neurons, shown by the presence of numerous varicose fibers immunostained for vasoactive intestinal peptide (VIP). These VIP fibers were poorly stained or unstained for synaptophysin, even after 7 weeks. At early post-operative times the VIP fibers grew irregularly; however, with longer post-operative times they appeared to target particular VIP-negative, noradrenergic neurons. Our results also indicate that some lumbar preganglionic axons regenerated during the post-operative period, although this only affected a minority of sympathetic neurons. These reinnervated sympathetic neurons were not associated with VIP fibers, suggesting that the new intrinsic connections may have precluded regeneration or targeting of preganglionic axons. Together these results demonstrate that there is considerable remodelling within pelvic ganglia after partial deafferentation. This occurs under conditions where spinal preganglionic axons can regenerate. New intra-ganglionic connectivity may be permanent and may impact on this regeneration.

Animals↗

Identification of the aromatic L-amino acid decarboxylase gene expression in various mice tissues and its modulation by immobilization stress in stellate ganglia.

Despite of the fact that the impact of various stressful stimuli on catecholamine biosynthetic enzyme gene expression, activity and immunoreactive protein has been intensively studied, less is known about the aromatic L-amino acid decarboxylase (AADC), the enzyme, which catalyzes decarboxylation of L-dihydroxyphenylalanine to dopamine. We focused on the identification of AADC mRNA and immunoprotein in various mice tissues and detected both in selected mice neuronal tissues (adrenal medulla, sympathetic stellate and cervical ganglia) and also in non-neuronal tissues (liver, spleen, kidney and all four parts of the heart). Surprisingly, although we failed to detect AADC mRNA in mice thymus, lungs and abdominal fat, we found presence of the AADC immunoprotein in lungs as well as in the abdominal fat. We also tested the hypothesis, whether single or repeated immobilization stress can affect the AADC mRNA or immunoprotein levels in mice stellate ganglia. We revealed that single immobilization stress exposure did not affect the AADC mRNA or immunoprotein levels, while repeated immobilization stress produced significant elevation of both, AADC mRNA and immunoprotein levels in stellate ganglia. The aromatic L-amino acid decarboxylase is generally not considered to be limiting in regulation of the catecholamine biosynthesis. However, our data suggest a possible participation of this enzyme in the regulation of catecholamine biosynthesis in stellate ganglia of repeatedly stressed mice.

Animals↗

Parkinsonism with basal ganglia lesions in a patient with uremia: Evidence of vasogenic edema.

Parkinsonian syndromes associated with basal ganglia pathology have very rarely been reported in patients with end-stage renal failure. The nature and pathophysiology of the basal ganglia lesion responsible for parkinsonism were unknown. A 48-year-old man who had advanced renal failure developed disturbance of balance and gait and decreased spontaneity. Brain magnetic resonance (MR) imaging disclosed bilateral basal ganglia lesions. By the finding of diffusion-weighted image, the apparent diffusion coefficient map, MR angiography, and SPECT, we suggest that the basal ganglia lesions may be the result of vasogenic edema attributable to focal hyperemia secondary to abnormal dilatation of small vessels.

Brain Edema↗

Sulfur-dioxide exposure increases TRPV1-mediated responses in nodose ganglia cells and augments cough in guinea pigs.

The objective of the present experiments was to study the effects of pulmonary inflammation induced by subacute Sulfur-dioxide (SO(2)) exposure on capsaicin-induced responses in isolated primary vagal sensory neurons and cough. Additionally, we examined the effects of SO(2) exposure on respiratory function and lung histology. All experiments were conducted 24 h after 4 days of subacute SO(2) (1000 ppm, 3 h/day for 4 days) exposure. In in vitro experiments, intracellular Ca(2+) concentrations were measured in single nodose ganglia cells isolated from SO(2) treated and control guinea pigs, using a fluorescence-based methodology. In nodose ganglia cells from SO(2)-exposed animals, intracellular Ca(2+) responses evoked by capsaicin (1 x 10(-7) and 1 x 10(-6) M) were significantly augmented (87% and 59%, respectively) compared to nodose ganglia from control animals. In vivo experiments, cough responses induced by a submaximal dose of aerosolized capsaicin (30 microM) were increased approximately 50% in SO(2) exposed animals compared to control animals. The enhanced cough response produced by SO(2) was inhibited by the corticosteroid, dexamethasone (10 mg/kg, p.o. b.i.d for 4 days and 10 mg/kg, p.o. once on day 5). In separate experiments, guinea pigs exposed to SO(2) displayed a decrease in respiratory frequency and minute ventilation and an increase in enhanced pause (PenH), a surrogate measure for pulmonary obstruction. Associated with the SO(2)-induced increase in cough and changes in respiratory parameters was an increase in BAL neutrophils. BAL neutrophil counts were 5+/-4 and 691+/-141 cells x 10(3)/ml for air and SO(2)-exposed animals, respectively. The neutrophillic inflammation induced by SO(2) was attenuated by dexamethasone treatment. Finally, staining for collagen, smooth muscle and goblet cells showed inflammation, remodeling and goblet cell metaphasia in the SO(2)-exposed animals. Our results demonstrate that SO(2) exposure enhances TRPV1 receptor function at the level of the nodose ganglia. This effect occurs in parallel with an increase sensitivity of the cough response to capsaicin.

Air Pollutants↗

Lesion of the subthalamic nucleus or globus pallidus does not cause chaotic firing patterns in basal ganglia neurons in rats.

The basal ganglia appears to play an important role in behavioral selection. One model (Berns and Sejnowski's) of basal ganglia function argues that the subthalamic nucleus plays a critical role in this selection process and predicts that the subthalamic nucleus prevents the basal ganglia and its re-entrant circuits with the thalamus and cerebral cortex from developing chaotic oscillations. We tested this prediction by generating three-dimensional sequential interval state space plots of the spike trains from 684 globus pallidus, substantia nigra pars reticulata and subthalamic neurons recorded in intact, subthalamic lesioned and globus pallidus lesioned rats, neurons which had previously been analyzed with more standard statistical methods. Only 1 neuron (a globus pallidus neuron in a subthalamic lesioned rat) of the 684 showed a chaotic attractor. In no case did subthalamic nucleus lesion induce a chaotic firing pattern elsewhere in the basal ganglia.

Action Potentials↗

Concurrent release of ATP and substance P within guinea pig trigeminal ganglia in vivo.

Neurons within sensory ganglia have been proposed to communicate via non-synaptic release of a diffusible chemical messenger, but the identity of the chemical mediator(s) remains unknown [J. Neurosci. 16 (1996) 4733-4741]. The present study addressed the possibility of co-released ATP and substance P (SP) within sensory ganglia to further advance the hypothesis of non-synaptic communication between sensory neurons. Microdialysis probes inserted into trigeminal ganglia (TRGs) of anesthetized guinea pigs were perfused with artificial cerebrospinal fluid and the collected perfusate analyzed for ATP and SP content using the firefly luciferin-luciferase (L/L) assay and radioimmunoassay, respectively. Significant reversible increases in ATP and SP levels were observed after infusion of 100 mM KCl or 1 mM capsaicin. Ca(2+)-free ACSF produced an eightfold increase in ATP levels, interpreted as a decrease in activity of Ca(2+)-dependent ecto-nucleotidases that degrade ATP. In contrast, KCl-induced release of ATP in the presence of normal Ca(2+) was blocked by Cd(2+), a voltage-gated Ca(2+) channel blocker, illustrating Ca(2+)-dependence of evoked ATP release. Since ganglionic release of ATP could arise from several neuronal and non-neuronal sources we directly tested acutely dissociated TRG neuron somata for ATP release. Neuron-enriched dissociated TRG cells were plated onto glass tubes and tested for ATP release using the L/L assay. Robust ATP release was evoked with 5 microM capsaicin. These data suggest that ATP is released concurrently with SP from the somata of neurons within sensory ganglia.

Adenosine Triphosphate↗

Ecto-nucleotidase of cultured rat superior cervical ganglia: dipyridamole is a novel inhibitor.

Based on studies of agonist potencies on intact rat superior cervical ganglia, it has been suggested that this ganglion possesses distinct receptors for purine and pyrimidine nucleotides. However, the potency of an agonist is dependent upon whether it is susceptible to extracellular metabolism by the tissue. The aim of this investigation was to study the metabolism of uridine or adenosine nucleotides and nucleosides and the effects of dipyridamole and an ecto-ATPase inhibitor ARL 67156 (6-N, N-diethyl-D-beta-gamma-dibromomethylene-ATP) on their metabolism. Adenosine- and uridine-5'-triphosphates (ATP and UTP) were catabolised by cultured rat superior cervical ganglia, to their di- and monophosphates. Both ATP and UTP breakdown was significantly inhibited by dipyridamole (10 mcM), whereas ARL 67156 (100 mcM), was a weaker inhibitor of ATP degradation and inhibited UTP breakdown by approximately 40%. Metabolism of ATP and UTP by cultured rat superior cervical ganglia was reduced after treatment with cytosine-beta-arabinoside, suggesting that non-neuronal cells along with neuronal cells contribute to their breakdown. In conclusion, these results indicate that rat superior cervical ganglia possess ecto-nucleotidases capable of catabolising purine and pyrimidine nucleotides to their nucleosides, and that dipyridamole is a potent inhibitor of ecto-nucleotidase activity.

5'-Nucleotidase↗

Transthoracic video endoscopic electrocautery of sympathetic ganglia for hyperhidrosis palmaris: special reference to localization of the first and second ribs.

BACKGROUND: The surgical technique for transthoracic endoscopic sympathectomy varies from one to three skin incisions, room air to carbon dioxide pneumothorax, and destruction of the second (T2), third (T3), and fourth sympathetic ganglia to destruction of the T2 ganglion only. A knowledge of the surgical anatomy of the apex may help the surgeon to safely use this technique. METHODS: Forty-seven patients with palmar hyperhidrosis underwent video-assisted endoscopic electrocautery of the T2 and T3 ganglia with the use of one-lumen endotracheal tube for general anesthesia, one skin incision, and carbon dioxide pneumothorax. Surgical anatomy, palm temperature, and surgical results were analyzed. RESULTS: The first ribs of 23 patients were endoscopically visible and most of these first ribs were not as parallel to the second ribs as the third ribs were. The first ribs of the remaining 24 patients were palpable with a diathermy bar. In all but three patients with dense pulmonary adhesions, the distal end of the intrathoracic segment of the subclavian artery was seen to pierce the pleura at the upper border of the first rib. Ninety-one palms remain dry and 27 patients develop compensatory sweating in an average follow-up of 12 months. Excluding three patients whose sympathetic ganglia could not be electrocauterized because of severe pulmonary adhesions, 95% of the remaining 44 patients are satisfied with the results. CONCLUSIONS: Transthoracic video endoscopic electrocautery of the T2 and T3 ganglia for patients with palmar hyperhidrosis may yield excellent results if the first rib can be properly identified.

Adolescent↗

Adrenocorticotropic hormone (MC-2) receptor mRNA is expressed in rat sympathetic ganglia and up-regulated by stress.

Stress triggered cardiovascular disorders are associated with elevated activity of the sympathetic nervous system, the major source of elevated plasma norepinephrine levels. Our previous studies revealed that administration of adrenocorticotropic hormone (ACTH) increases the gene expression of norepinephrine biosynthetic enzymes and several neuropeptides in rat sympathetic ganglia as much as stress. Here, we examine whether an ACTH-responsive receptor is expressed in rat superior cervical (SCG) and stellate ganglia (StG). Using reverse transcriptase-polymerase chain reaction (RT-PCR) we found expression of MC-2 receptor mRNA in these ganglia. Identical DNA fragments were amplified with mRNA from SCG, StG or from adrenal cortex. Sequencing revealed extensive homology to published sequences of mouse and human MC-2 receptor. Real time PCR was used to quantitate MC-2 receptor mRNA levels in the SCG under basal conditions and following immobilization stress. Immobilization stress triggered a large increase in MC-2 receptor mRNA in SCG. The results provide the first evidence that rat sympathetic ganglia express MC-2 receptor gene and are a target tissue for the peripheral actions of ACTH in response to stress.

Animals↗

Innervation of guinea-pig stellate ganglia by nitric oxide synthase, cocaine- and amphetamine-regulated transcript protein- and pituitary adenylate cyclase activating polypeptide-immunoreactive fibers.

The present study analyzed using immunohistochemical labeling the distribution and co-localization of nitric oxide synthase (NOS), cocaine- and amphetamine-regulated transcript peptide (CARTp) and pituitary adenylate cyclase activating polypeptide (PACAP) with choline acetyltransferase (ChAT)-immunoreactive fibers in the guinea-pig stellate ganglia. ChAT-immunoreactive fibers make pericellular baskets around virtually all stellate ganglia neurons. Pericellular baskets of NOS, CARTp and PACAP fibers were also present around numerous stellate ganglia neurons. Although all the NOS and PACAP fibers also exhibited ChAT immunoreactivity, only some of the CARTp fibers were ChAT-immunoreactive. No evidence of co-localization of NOS, PACAP and CARTp was obtained.These results indicate that NOS, PACAP and CARTp are present in distinct preganglionic axons innervating the guinea-pig stellate ganglia.

Animals↗

Immunoreactive localisation of P2Y1 receptors within the rat and human nodose ganglia and rat brainstem: comparison with [alpha 33P]deoxyadenosine 5'-triphosphate autoradiography.

The present study employed standard peroxidase immunohistochemistry to map the distribution of P2Y(1) receptors in the rat brainstem and nodose ganglia and characterised the binding profile of [alpha(33)P]dATP. Binding of [alpha(33)P]dATP was fully displaceable by adenosine 5'-triphosphate (ATP), and was found on both human and rat nodose ganglia, and throughout the rat brainstem, including the nucleus tractus solitarius and ventrolateral medulla. [Alpha(33)P]dATP binding in the human nodose ganglia was significantly displaced by both 2-methylthio ATP and alpha,beta-methylene ATP, but not by uridine 5'-triphosphate, pyridoxalphosphate-6-azophenyl-2',4'-disulfonic acid, 8,8'-(carbonylbis(imino-4,1-phenylenecarbonylimino-4,1-phenylenecarbonylimino))bis(1,3,5-naphtalenetrisulfonic) acid (NF279) or N-ethylcarboxamidoadenosine. [Alpha(33)P]dATP binding in the rat nodose ganglia and brainstem was significantly displaced by only 2-methylthio ATP, suggesting that [alpha(33)P]dATP is binding to P2Y receptors in the rat. Binding of [alpha(33)P]dATP was also significantly displaced by alpha,beta-methylene adenosine 5'-diphosphate, suggesting a component of the binding is to endogenous ecto-5'-nucleotidase, however, almost all binding could be displaced by a combination of receptor agonists (2-methylthio ATP, uridine 5'-triphosphate and alpha,beta-methylene ATP), suggesting preferential binding to receptors. Immunoreactivity to P2Y(1) receptor (P2Y(1)-IR) exhibited similar distribution patterns to [alpha(33)P]dATP binding, with a clear topographic profile. Particularly dense P2Y(1)-IR labeling was evident in cells and fibres of the dorsal vagal complex. Immunolabeling was also present in the dorsal motor nucleus of the vagus and nucleus ambiguus, indicating the possibility of P2Y(1) receptors on vagal efferents. Unilateral vagal ligation was also performed to examine the transport of P2Y(1) receptor, using both immunohistochemistry and [alpha(33)P]dATP autoradiography. Accumulations of both P2Y(1)-IR and [alpha(33)P]dATP binding were apparent adjacent to both ligatures, suggesting bi-directional transport of P2Y(1) receptors along the rat vagus nerve. This current study represents the first description of P2Y(1) receptor distribution within the rodent brainstem and nodose ganglion and also characterises [alpha(33)P]dATP binding to P2Y receptors.

Adolescent↗