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Ethanol-induced taste aversions: lack of involvement of acetaldehyde and the area postrema.

Two experiments were run to evaluate the role of acetaldehyde and the area postrema in the acquisition of an ethanol-induced conditioned taste aversion. An ethanol-induced taste aversion was observed in male Sprague-Dawley rats with a dose of 4 g/kg. PO, but not after doses of 1 or 2 g/kg. Pretreatment with 4-methylpyrazole (8 mg/kg, IP), which itself did not induce an aversion as compared to pyrazole (68 mg/kg, IP) that did, and/or prior application of lesions of the area postrema had no influence on the development of an ethanol-induced taste aversion. The results indicate that ethanol-induced taste aversion learning does not result from the metabolism of ethanol to acetaldehyde and does not, like other toxins, involve the mediation of the area postrema.

Acetaldehyde↗

Systemic glutamate induces degeneration of a subpopulation of tyrosine hydroxylase-immunoreactive neurons in the rat area postrema.

Neuronal damage in the area postrema (AP) of 12-14-week-old male rats was induced by subcutaneous administration of monosodium glutamate (MSG). An immunocytochemical method was used to visualize catecholaminergic neurons in the AP after MSG-treatment. Some tyrosine hydroxylase-immunoreactive neurons exhibited marked signs of degeneration, while others appeared undamaged. We conclude that catecholamine-synthesizing neurons in the AP are differentially sensitive to the neuroexcitotoxic effect of systemic glutamate.

Animals↗

Enhanced saline intake and decreased heart rate after area postrema ablations in rat.

The area postrema (AP) is a circumventricular organ located in the dorsal medulla. Previous studies found that AP lesions lead to increased saline ingestion in the rat. The salt appetite was thought to be a result of primary disruptions in sodium regulation or in cardiovascular regulation. To assess this we measured food and fluid intakes, urinary electrolyte and aldosterone concentration, and blood pressure and heart rates in AP lesioned and control animals during a period of normal sodium intake and during a period of excessive sodium intake. Rats with AP lesions exhibited sodium appetite but not natriuresis. In fact, sodium intake greatly exceeded output. Their urinary aldosterone levels were similar to those of control animals during both periods. The lesioned rats also had lowered heart rates, yet, their blood pressures were similar to control animals. These results are discussed with reference to a possible role of the AP in satiety and in maintaining homeostasis.

Aldosterone↗

The connectivity of the area postrema in the ferret.

The area postrema (AP) is the chemosensitive trigger zone for the emetic reflex. We have investigated the connectivity of the AP and adjacent solitary complex (SC) to identify possible sites of the motor emetic center. The AP and SC were infused with HRP or WGA-HRP in 30 ferrets that were perfused transcardially after 24-72 h. A block from the pons to upper cervical spinal cord, and one with hypothalamus and basal forebrain, was cut at 50 microns, reacted, and mounted. Data support the conclusion, at variance with those from other preparations, that in ferrets the AP has reciprocal connections only with the SC, which serves as a relay in both ascending and descending pathways between AP and higher levels of the neuraxis. Connectivity of the SC with brain stem and forebrain structures including the rostral ventrolateral medulla, parabrachial nuclei, paraventricular nucleus, and amygdala was demonstrated. At least in ferrets, our results suggest that the motor emetic center must be located within the SC. While this may not apply to all species, it is also possible that some reports of AP projections elsewhere were results of label within the SC. Alternatively, the somewhat different pattern of emesis in the ferret as compared to the dog (greater role for vagal inputs in response to radiation and cytotoxic drugs, lesser role for humoral inputs) may reflect differences in AP connectivity.

Animals↗

Cardiovascular responses to microinjections of excitatory amino acids into the area postrema of the rat.

Although, area postrema (AP) as been implicated in the regulation of cardiovascular function, there is no consensus regarding the type of responses elicited by stimulation of this brain structure. Microinjections (50 nl) of smaller concentrations of excitatory amino acid receptor agonists (e.g., NMDA, KA and trans-ACPD, 10 microM each) into the AP elicited pressor and tachycardic responses in unanesthetized decerebrate as well as urethane-anesthetized rats. Microinjections of higher concentrations (e.g., 50 microM NMDA) of excitatory amino acids (EAAs) into the AP elicited an initial pressor and tachycardic response which was followed by a depressor and bradycardic response; when high concentrations of NMDA were microinjected into the AP, enough concentration may have reached the nucleus tractus solitarius (nTS) to elicit depressor and bradycardic responses. Alternatively, high concentrations of NMDA may excite known projections from AP to the nTS.

2-Amino-5-phosphonovalerate↗

Non-NMDA and NMDA receptors in the synaptic pathway between area postrema and nucleus tractus solitarius.

Area postrema (AP) modulates cardiovascular function through excitatory projections to neurons in nucleus tractus solitarius (NTS), which also process primary sensory (including cardiovascular-related) input via the solitary tract (TS). The neurotransmitter(s) and their receptors in the AP-NTS pathway have not been fully characterized. We used whole cell recordings in voltage- and current-clamp modes in the rat brain stem slice to examine the role of ionotropic glutamatergic receptors and alpha2-adrenergic receptors in the pathway from AP to NTS neurons receiving visceral afferent information via the TS. In neurons voltage clamped at potentials from -100 to +80 mV, AP stimulation (0. 2 Hz) evoked excitatory postsynaptic currents having a fast component blocked by the non-N-methyl-D-aspartate (NMDA) receptor antagonist 1,2,3,4-tetrahydro-6-nitro-2, 3-dioxobenzoquinoxaline-7-sulfonamide (NBQX; 3 microM, n = 7) and a slow component blocked by the NMDA receptor antagonist DL-2-amino-5-phosphonovaleric acid (APV; 50 microM, n = 8). Although NBQX (3 microM, n = 14) abolished AP-evoked action potentials, APV (50 microM, n = 9 or 500 microM, n = 6) or yohimbine, (200 nM, n = 5 or 2 microM, n = 10) did not. Thus, although AP stimulation activates both non-NMDA and NMDA receptors on NTS neurons receiving TS input, only non-NMDA receptors are required for synaptic transmission.

2-Amino-5-phosphonovalerate↗

Lateral parabrachial lesions attenuate ingestive effects of area postrema lesions.

Lesions of the area postrema and adjacent nucleus of the solitary tract (AP lesions) cause rats to consume increased amounts of palatable food in short duration tests. Because the lateral parabrachial nucleus (lPBN) receives a prominent afferent projection from the AP and adjacent nucleus of the solitary tract, it is possible the lPBN plays a role in the altered ingestive behaviors observed in AP-lesioned rats. The present study examines the role of the lPBN in overingestion of highly palatable foods subsequent to AP lesions. We found that lesions of the lPBN alone did not cause rats to consume increased amounts of palatable food. Rather, when lPBN lesions were produced before AP lesions, increased intake of highly palatable food did not occur. Moreover, when AP-lesioned rats received subsequent lPBN lesions, the previously established overingestion of palatable foods was abolished. These results indicate that the lPBN is necessary in the pathogenesis of AP lesion-induced overingestion of highly palatable foods.

Animals↗

Vasopressin actions on area postrema neurons in vitro.

The area postrema (AP) is a circumventricular organ located on the dorsal surface of the medulla. Substantial evidence suggests that the AP is an important site involved in cardiovascular regulation. Arginine vasopressin (AVP) is thought to act at the AP to increase the sensitivity of the baroreceptor reflex. We have therefore examined the effects of AVP on AP neurons with the use of extracellular single unit recordings in vitro. Coronal medullary brain slices (thickness = 400 microns) were obtained from male Sprague-Dawley rats and maintained in oxygenated artificial cerebrospinal fluid (aCSF). The slices were perfused with AVP (10(-8) to 10(-6) M), and the effect on single AP neurons was recorded. A total of 79 AP neurons was tested of which 50 (63.3%) were excited by AVP and 5 (6.3%) were inhibited, whereas the remaining 24 (30.3%) cells were unaffected. The excitatory effects of AVP were dose dependent: firing rate increased 92.6 +/- 25.8% at 10(-8) M, 289.4 +/- 53.9% at 10(-7) M, and 456.8 +/- 113.1% at 10(-6) M, respectively. We also examined whether these effects of AVP resulted from direct actions of this peptide on AP cells by testing if responses were retained during blockade of synaptic transmission (achieved by perfusion with a low Ca(2+)-high Mg2+ aCSF) in 11 cells excited by AVP. Nine of these cells were excited by AVP during such synaptic blockade. Finally, we demonstrated that the excitatory responses of five AP cells to AVP were all totally abolished by perfusion of slices with aCSF containing the V1 antagonist ([1-beta-mercapto-beta,beta-cyclopentamethylene propionic acid,2-(O-methyl)tyrosine]-Arg8-vasopressin; Peninsula Laboratories, 10(-6) M).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Efferent projections of the area postrema demonstrated by autoradiography.

The efferents connections of the area postrema (AP) have been studied autoradiographically following iontophoretic injections of 3H-glycine or 3H-leucine into the area postrema. Precise control of the diffusion of the labelled amino acids injected iontophorectically into the AP was made using the technique of sectioning with a cryostat. AP projects to a great number of structures. Projections to nucleus tractus solitarius (NFS), dorsal vagal nucleus, nucleus intercalatus, nucleus praepositus hypoglossi, nucleus hypoglossal, the mesencephalic nucleus of V nerve, locus coeruleus and superior and inferior colliculi are shown bilaterally. The density of the efferents was greatest to the NFS and the LC. Corelations are suggested with functional mechanisms of cardiovascular regulation.

Animals↗

Chemical stimulation of the area postrema induces cardiorespiratory changes in the cat.

The purpose of our study was to determine the cardiorespiratory effects of exciting cell bodies of the area postrema of the cat. This was accomplished by local application of L-glutamic acid (bilateral application of 5 microliter of a 250-1000 mM solution) and kainic acid (bilateral application of 5 microliter of a 40 mM solution) to the area postrema of chloralose-anesthetized cats while monitoring arterial pressure, heart rate, tidal volume and respiratory rate. These excitatory amino acids activate neuronal cell bodies but not axons of passage. L-Glutamic acid produced a dose-dependent increase in arterial pressure, decreases in respiratory rate and minute volume and, occasionally, ventricular tachyarrhythmias. Kainic acid produced effects similar to those seen with L-glutamic acid except the changes in respiratory activity were more pronounced with each animal exhibiting respiratory arrest. In artificially respired animals, kainic acid produced similar cardiovascular changes as those occurring in spontaneously breathing animals (i.e. increases in arterial pressure of 61 +/- 5.7 mm Hg, and in heart rate of 32 +/- 8.3 beats/min). Finally, application of kainic acid to the area postrema abolished the pressor and tachycardic responses to bilateral occlusion of the carotid arteries. These results suggest that activation of cell bodies in the area postrema can result in pronounced cardiorespiratory changes.

Animals↗

Motor activity changes and conditioned taste aversions induced by administration of scopolamine in rats: role of the area postrema.

Three experiments examined the effects of centrally and peripherally acting scopolamine (scopolamine hydrochloride-SHC) or only peripherally acting scopolamine (scopolamine methyl nitrate-SMN), on motor activity levels and the ability of these agents to induce taste aversions. In Experiment 1 rats were injected with isotonic saline, 1 mg/kg SHC, or 1 mg/kg SMN. SHC produced significant increases in stabilimeter activity (p less than 0.025) and in rearing response frequency (p less than 0.01), whereas SMN resulted in significantly less activity (p less than 0.025). Both agents induced strong conditioned taste aversions to saccharin (p less than 0.01). Experiment 2 examined the role of the area postrema in mediating these drug induced behavioral changes. Sham lesioned and area postrema lesioned rats were given saline, SHC or SMN (1 mg/kg for both drugs) and examined for changes in activity, rearing response frequency and induction of taste aversions to saccharin. SHC again produced significant increases in activity (p less than 0.01), whereas SMN produced decrements in activity (p less than 0.05). However, the brain lesion did not consistently alter the effects of these drugs on activity but it did reduce the amount of the decrement observed in rearing responses in SMN treated rats. The brain lesion also altered the ability of the drugs to induce taste aversions. Both SMN and SHC produced strong taste aversions in the sham lesioned rats (p less than 0.01) but no significant aversions were observed in the area postrema lesioned rats. Experiment 3 examined the ability of the brain lesion to alter the effects of SMN by using a within groups design. Area postrema lesions were found to attenuate but not abolish, the inhibitory effects of SMN on both activity levels and rearing responses (p less than 0.03). The results of these experiments suggest that in the absence of the chemically-sensitive area postrema both SMN and SHC fail to induce taste aversions and the inhibitory effect of SMN on spontaneous activity is attenuated.

Animals↗

The area postrema plays no role in the pressor action of angiotensin in the rat.

The area postrema has been shown to have a major role in mediating the pressor effects of peripheral angiotensin in the dog, cat, and rabbit. The purpose of this study was to ascertain the function of the medullary circumventricular structure in the conscious rat. The pressor potency of angiotensin administered into the vertebral and carotid arteries was compared with intra-aortic infusions of angiotensin. Although no difference in pressor activity of angiotensin could be detected between intraaortic and intravertebral administration, greater sensitivity was observed during intracarotid infusion. No difference in the course of one-kidney renal hypertension was observed between sham-lesioned rats and animals with an area postrema lesion. In addition, lesioned and sham-lesioned animals showed equivalent responses to graded doses of angiotensin administered either intravenously or into the lateral ventricle. It was concluded that in the rat the area postrema plays no role in mediating the central nervous system actions of angiotensin whether the peptide reaches the brain via the blood or the cerebrospinal fluid.

Angiotensin II↗

Central opiate system modulation of the area postrema pressor pathway.

Angiotensin II, when given into the vertebral arteries, acts at the area postrema to augment central sympathetic vasomotor activity. The mechanism of action is unknown but recent evidence implicates an interaction with the opiate system. In dogs anesthetized with chloralose either alone or in combination with morphine, naloxone blunted the pressor response to vertebrally administered angiotensin II by 50%. Addition of morphine to dogs anesthetized with chloralose only doubled the pressor response to identical doses of angiotensin II. On the other hand, the magnitude of the pressor responses to intravenously infused angiotensin II were unaltered by either naloxone or morphine. Likewise, responses to norepinephrine given vertebrally and intravenously were not similarly affected. Therefore, naloxone-induced changes in vascular responsiveness were not responsible for the altered sensitivity of the area postrema to angiotensin II following blockade of endogenous opiates. The data suggest that there exists a previously unrecognized interaction of the endogenous opiate system in the medulla in mediating the pressor effects of angiotensin II at the level of the area postrema.

Angiotensin II↗

Glucose-responsive neurons exist within the area postrema of the rat: in vitro study on the isolated slice preparation.

Responses to glucose of spontaneously active neurons were investigated by extracellular recording in the rat area postrema slice preparations (in vitro). Among 67 spontaneously active neurons, 16 neurons displayed a marked increase or decrease in discharge rate in response to increases or decreases of the glucose concentration in perfusate. These results confirm the existence of glucose-responsive neurons within the area postrema suggested in prior in vivo experiments. Response to CCK or dopamine was also examined on the isolated area postrema slices. The neuron that showed a marked increase in discharge rate responding to glucose elicited a marked increase of discharge rate in response to 2.1 microM CCK, suggesting that glucose and CCK affect the same neurons. Some neurons showed a marked increase or decrease in the discharge rate in response to 20 microM dopamine, but these neurons showed neither response to CCK nor to glucose. It is likely that different neuronal networks in the area postrema contribute to control of ingestion and to initiation of nausea.

Animals↗

[The role of tachykinin NK-1 receptors in emetic action in the area postrema of ferrets].

Substance P (SP) is a member of the tachykinin family of bioactive peptides and has highest affinity for the NK-1 receptor. We have developed the non-peptide compound HSP-117 as a selective antagonist of the NK-1 receptor. Binding of 3H-SP to the membranes of IM-9 cells was inhibited by the antagonists HSP-117 and CP-99,994, the inhibitory activity of HSP-117 being 50-fold that of CP-99,994. The SP-induced firing responses of single neuron activity in slices of the nucleus tractus solitarius of ferrets were inhibited by 10 microM HSP-117. Intracerebroventricular injection of HSP-117 significantly inhibited retching and vomiting induced by copper sulphate and morphine and the inhibitory effect of HSP-117 on emesis was greater than that of CP-99,994. Moreover, emesis induced by copper sulphate and morphine were inhibited by the microinjection of HSP-117 and CP-99,994 into the area postrema and by lesion of the area postrema. These results indicate that HSP-117 is a potent anti-emetic agent, blocking NK-1 receptors in the area postrema and that NK-1 receptors in the area postrema play an important role in emesis induced by broad-spectrum emetic stimuli.

Animals↗

Area postrema: cholinergic and noradrenergic regulation of emesis. A new concept.

In unanaesthetized cats the biochemical mechanisms and the functional characteristics of the emetic action of injection of noradrenaline and McN-A-343, a ganglionic muscarinic stimulant into the cerebral ventricle (i.c.v.) through chronically implanted cannulae were investigated. Both produced dose-dependent and shortlasting emetic response. The emesis evoked by noradrenaline was abolished, whereas the emesis induced by McN-A-343 was not completely blocked after ablation of the area postrema. Further, the emetic response to noradrenaline as well as to McN-A-343 was attenuated or blocked in cats pretreated with 6-hydroxydopamine (i.c.v.) and hemicholinium (i.c.v.); it was abolished in cats pretreated with reserpine (i.c.v.). On the other hand, the emetic response to i.c.v. noradrenaline and to i.c.v. McN-A-343 was not virtually altered in cats pretreated with bretylium (i.c.v.), alpha-methyl-p-tyrosine (i.c.v.) and 5,6-dihydroxytryptamine (i.c.v.). It is postulated that noradrenergic neurones as well as cholinergic axon terminals within the area postrema are necessary for the emetic action of noradrenaline, whereas cholinergic axon terminals within the area postrema subserve the emetic response to McN-A-343. A functional link between cholinergic terminals and noradrenergic neurones as well as a modulatory role of noradrenergic afferents on cholinergic afferents mediating emesis within the area postrema is further proposed. Thus, noradrenergic neurones might represent a common site of confluence of different inputs subserving the emesis in the area postrema. Finally, cholinergic terminals sometimes bypass this area and synapse in the emetic regions of the brainstem regulating emesis.

(4-(m-Chlorophenylcarbamoyloxy)-2-butynyl)trimethy↗

Characterization of orexin A immunoreactivity in the rat area postrema.

The distribution of orexin A immunoreactivity and the synaptic relationships of orexin A-positive neurons in the rat area postrema were studied using both light and electron microscopy techniques. At the light microscope level, numerous orexin A-like immunoreactive fibers were found within the area postrema. Using electron microscopy, immunoreactivity within fibers was confined primarily to the axon terminals, most of which contained dense-cored vesicles. Both axo-somatic and axo-dendritic synapses made by orexin A-like immunoreactive axon terminals were found, with these synapses being both symmetric and asymmetric in form. Orexin A-like immunoreactive axon terminals could be found presynaptic to two different immunonegative profiles including the perikarya and dendrites. Occasionally, some orexin A-like immunoreactive profiles, most likely to be dendrites, could be seen receiving synaptic inputs from immunonegative or immunopositive axon terminals. The present results suggest that the physiological function of orexin A in the area postrema depends on synaptic relationships with other immunopositive and immunonegative neurons, with the action of orexin A mediated via a self-modulation feedback mechanism.

Animals↗

Enkephalin-like immunoreactivity in rat area postrema: ultrastructural localization and coexistence with serotonin.

The ultrastructure of enkephalin-containing neurons and their capacity to take-up [3H]serotonin were examined in the area postrema. Untreated adult rats and rats with intraventricular infusions of 10(-4) M tritiated serotonin, 5-hydroxytryptamine [( 3H]5-HT) were perfused with 4% paraformaldehyde and 0.2-0.5% glutaraldehyde. Coronal Vibratome sections through the area postrema from both groups of animals were immunocytochemically labeled with an antiserum to leucine Leu5-enkephalin. The sections from the animals infused with the isotope subsequently were processed for autoradiography. Enkephalin-like immunoreactivity (ELI) was detected in perikarya, dendrites, axons and axon terminals most frequently located along the ventricular and ventrolateral portions of the area postrema. The labeled perikarya were few in number and were characterized by a thin rim of cytoplasm containing peroxidase immunoreactivity. Dendrites and terminals containing ELI formed synapses primarily with unlabeled axon terminals and dendrites, respectively. However, terminals containing ELI also formed synaptic junctions with other unlabeled axon terminals. Appositions between enkephalin-containing processes and modified glia were occasionally seen near the ventricular surface. In sections processed for both immunocytochemistry and autoradiography, approximately 5% of the terminals containing ELI showed uptake of [3H]5-HT. We conclude that neurons containing ELI are primarily, but not exclusively, associated with other intrinsic neurons or afferents in the rat area postrema and that some of the enkephalin-labeled terminals have the capacity to take-up serotonin. Specificity of uptake of [3H]5-HT in neurons containing endogenous serotonin and factors which may contribute to the low probability of detecting both peroxidase and autoradiographic markers in single sections are discussed.

Animals↗