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Area postrema is critical for angiotensin-induced hypertension in rats.

The effect of surgical ablation of the area postrema on acute (5-10 minutes) and chronic (5-10 days) increases in mean arterial pressure produced by intravenous infusion of angiotensin II in conscious, instrumented rats was studied. In agreement with previous studies, pressor responses of area postrema-ablated rats (n = 11) to acute angiotensin II infusion were identical to those of control sham-lesioned rats (n = 13). In these same rats, however, a 5-day infusion of angiotensin II produced a sustained hypertension in the sham-lesioned group whereas mean arterial pressure was increased only transiently (1-3 days) in the area postrema-ablated rats. No differences before infusion of arterial pressure, heart rate, water intake, urinary sodium excretion, and urinary potassium excretion were observed between sham-lesioned and area postrema-ablated rats; only arterial pressure was changed significantly during angiotensin II infusion in either group. Twenty-four hours after terminating angiotensin II infusion, mean arterial pressure was within the normotensive range in both sham-lesioned and area postrema-ablated rats. In a separate group of sham-lesioned (n = 13) and area postrema-ablated (n = 12) rats, angiotensin II was infused intravenously for a 10-day period; mean arterial pressure was increased significantly over the entire 10-day infusion in sham-lesioned rats, but for only 1 day in area postrema-ablated rats. An intact area postrema appears necessary for the development of chronic, but not acute, hypertension during intravenous infusion of angiotensin II in the rat.

Angiotensin II↗

Ablation of the area postrema and emesis.

The emetic action of dopamine, norepinephrine, epinephrine, nicotine, dimethylphenyl-piperazinium (DMPP), and 4-m-chlorophenylcarbamoyloxy-2-butynyltrimethylammonium (McN-A-343) injected intracerebroventricularly (i.c.v.) to the unanesthetized cat was investigated and compared. ED50 values (mg) were as follows: nicotine, 0.011; epinephrine, 0.047; norepinephrine, 0.57; DMPP, 0.9; dopamine, 1.66; and McN-A-343, 4.42. The most potent was nicotine, whereas the least active McN-A-343. On the other hand, DMPP produced the longest emetic response, about 30 min, while McN-A-343-induced emesis lasted up to 1 min. The ablation of the area postrema abolished the emetic response to i.c.v. dopamine, norepinephrine, epinephrine, nicotine, and DMPP. However, the emetic response to i.c.v. McN-A-343 was significantly reduced in cats with an ablated area postrema. Taken together, the results obtained show that the area postrema is almost always involved in the central regulation of emesis and that the area postrema represents, in most cats, a common site of confluence of different inputs subserving the emesis.

Animals↗

Angiotensin II binding in area postrema and nucleus tractus solitarius of SHR and WKY rats.

The distribution of angiotensin II (AII) binding sites in the area postrema (AP) and adjacent nucleus tractus solitarius (NTS) was compared in spontaneously hypertensive (SHR) rats and normotensive Wistar-Kyoto (WKY) rats. 125I[Saralasin-1-Isoleucine8]-Angiotensin II (125I[SI]-AII) binding density was quantitated from autoradiographic images by computer-assisted image analysis. Seventeen 30 microns serial coronal sections inclusive of the entire AP were analyzed as either individual sections or as groups of sections designated as caudal, middle, or rostral area postrema regions. 125I[SI]-AII binding density was greatest in caudal AP and declined progressively in the rostral direction in both strains; however, binding density in SHR was significantly higher than in WKY rats at each level of the AP analyzed. 125I[SI]-AII binding in the entire area postrema was approximately 46% higher in SHR rats. In the NTS, however, there were no differences in binding density between strains. At the middle level of the AP, 125I[SI]-AII binding was highest in the ventral midline and lowest in the dorsolateral region. In the NTS, the highest 125I[SI]-AII binding density was found in the pars commissuralis and pars medialis. In conclusion a) 125I[SI]-AII binding density was nonhomogeneous in the AP and NTS of both strains, b) qualitatively similar patterns of nonhomogeneity of binding in the AP and NTS were noted in both strains; however, c) the SHR strain consistently had higher density AII binding than WKY in AP, but not in NTS.

Angiotensin II↗

Fine structure of ependymal cysts in and around the area postrema of the rat.

Peculiar cells forming cysts were observed in the area postrema and sometimes also in the choroid plexus and the tela chorioidea near the area postrema, and were studied in detail by electron microscopy. The cytological features of the cyst cell and its junctional relationship to neighboring cells imply that cyst cells are derived from ependymal and choroid epithelial cells. The cyst cells usually contact directly the perivascular spaces of postremal, choroidal or pial capillaries, where the cytoplasm is often considerably attenuated. The cystic lumen is commonly filled with a flocculent material. The limiting membrane of the cystic lumen, which frequently bears cilia and microvilli, has the same thickness as the surface cell membrane. In many cases the cyst is surrounded by the cytoplasm of a single cell. In some cases, however, two cells participating in the formation of the cyst, although one is only a slender process and joined by a zonula occludens with the main cyst cell. Horseradish peroxidase (HRP) injected into the cerebrospinal fluid (CSF) space failed to enter the cystic lumen. A possible significance of the cyst in relation to the CSF and blood circulation was considered.

Animals↗

Systemic angiotensin II does not act at the area postrema to suppress alcohol intake.

The area postrema (AP), a circumventricular organ located in the dorsal medulla, is involved in the coordination of visceral and taste sensory stimuli in the control of ingestive behaviors. This area lacks a blood-brain barrier and is rich in angiotensin receptors. Because systemically administered angiotensin II reduces voluntary alcohol intake, we examined the effect of AP lesions on alcohol intake and on the ability of ANG II reduces alcohol intake. Compared to sham-operated controls, rats with AP lesions consumed significantly more of a palatable 3% alcohol solution, and equal amounts of a less palatable 6% alcohol solution, but the lesion did not alter the ability of ANG II to reduce alcohol intake. These findings indicate that while the AP plays no role in the suppressive effect of ANG II on alcohol consumption, it can modulate alcohol consumption through its responsiveness to orosensory stimuli.

Alcohol Drinking↗

Reciprocal synaptic relations between enkephalinergic and GABAergic neurons in the area postrema of the rat.

A preembedding, double immunostaining technique was used to study synaptic relations between enkephalinergic and GABAergic neurons in the area postrema of the rat. As a main result, the enkephalinergic dendrites received many synapses from GABAergic axon terminals, and most of the synapses were symmetrical. Enkephalinergic neuronal perikarya received a few synapses from GABAergic axon terminals, and a few enkephalinergic axon terminals were found presynaptic to GABAergic neurons. Synapses between enkephalinergic profiles were frequent, but no axo-axonic synapses were seen. These findings suggest that GABAergic innervation of enkephalinergic neurons is the main relation between the two kinds of neurons in the area postrema. The synapses between the enkephalinergic axon terminals and GABAergic neurons might be explained as being part of the local servo system of the area postrema.

Animals↗

Electron microscopic immunocytochemical localization of substance P in the area postrema of rat.

Electron microscopic immunocytochemistry was employed to examine the ultrastructure of processes containing substance P-like immunoreactivity (SP-LI) in the area postrema of rat. SP-LI was present within axons and axon terminals which formed synapses with dendrites of intrinsic neurons in the rat area postrema. Specialized contacts between labeled processes and perivascular or ependymal glia cells were not observed. These data suggested that substance P may act as a neurotransmitter involved primarily with neuronal (axodendritic) interactions within the area postrema.

Animals↗

Area postrema excites and inhibits sympathetic-related neurons in rostral ventrolateral medulla in rabbit.

This study investigated the effects of area postrema stimulation on the activity of cardiovascular-related neurons in the rostral ventrolateral medulla and determined whether the effects were mediated by cell bodies. Results are based on recordings of extracellular spikes from 113 neurons in 37 alpha-chloralose- or pentobarbital sodium-anesthetized rabbits. Single sequential stimuli evoked an excitation (onset, 22 +/- 8 ms; duration, 20 +/- 14 ms) followed by an inhibition (onset, 53 +/- 21 ms; duration, 127 +/- 82 ms) in 1) 58 neurons with discharge patterns that were correlated with sympathetic nerve activity (determined by spike-triggered averaging) and with the cardiac cycle (determined by post-R wave-triggered histograms) and that were inhibited by increasing arterial pressure and 2) 27 neurons that exhibited a cardiac rhythm but not a sympathetic rhythm. Area postrema-evoked excitation and inhibition were markedly attenuated by kainic acid injections in area postrema, suggesting that both inputs were derived from cell bodies. The results suggest that area postrema neurons may modulate the activity of medullary neurons in the baroreflex-sympathetic arc as well as neurons in other networks that share baroreceptor input but may not be related to sympathetic nerve activity.

Animals↗

Lesion of the area postrema region attenuates hypertension in spontaneously hypertensive rats.

To determine whether the area postrema contributes to the development of hypertension in spontaneously hypertensive rats (SHR), sham or electrolytic lesions of the area postrema (AP) were made in 4-week-old SHR and Wistar-Kyoto (WKY) controls. From weeks 5 through 16, systolic pressure was measured via tail plethysmography. While blood pressure rose markedly in sham-operated SHR, increases in pressure were small in AP-ablated SHR and similar to those seen in all WKY. Subsequent direct measurements of mean arterial pressure in the same rats showed a significant correlation (r = 0.87, p less than 0.01) with the pressure data acquired via weekly tail-cuff measurement, thereby confirming that hypertension in AP-ablated SHR had indeed been attenuated. Analysis of several hundred computer-acquired measurements of mean arterial pressure from each rat showed that AP ablation shifted the distribution of mean arterial pressure to a lower range in SHR but not WKY. Ablation of the AP also decreased resting heart rate in SHR but not WKY. Suppression of heart rate in response to intravenous phenylephrine was equivalent in sham-operated and AP-ablated rats, suggesting that baroreflex-mediated slowing of heart rate was not impaired. In response to intravenous angiotensin II, suppression of heart rate was similar in sham and AP-ablated SHR, and actually was enhanced in AP-ablated WKY. Histological evaluation of the lesions indicated that visible damage to the adjacent nuclei of the solitary tracts was confined to a small portion of the commissural nucleus.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

The effect of removing area postrema on the sodium and potassium balances and consumptions in the rat.

Previous studies have indicated that rats lose weight and develop a taste preference for solutions of sodium chloride after area postrema has been removed; results in other species have suggested that area postrema may be concerned with regulating the excretion of sodium and potassium. The results reported here demonstrate that in rats the reduction of weight can be explained adequately by anorexia, that diminished excretion of sodium and potassium results from anorexia, and that the consequent slower rate of gain of weight causes diminution of sodium and potassium balances. The results also show that the rats do not drink more of a solution of sodium chloride because they eat less, and conversely that when they can drink a saline solution they do not eat more: and that they do not drink more saline because of salt loss. Furthermore, individual rats show no significant correlation between the severity of anorexia, and the increased consumption of saline. Taken together the results suggest that primary disorder of regulation of intake occurs in rats after area postrema has been removed, and that the reduced intake of food and the increased intake of sodium chloride occur independently.

Animals↗

Imiquimod-elicited emesis is mediated by the area postrema, but not by direct neuronal activation.

The immunomodulator, imiquimod, has been shown to have antiviral and antitumor properties in animal models. It also has been reported to alter cytokine levels in both animals and humans. However, because imiquimod appeared to be emetic, studies were conducted to determine the degree of sensitivity, and the pathways involved. Subcutaneous administration of > or = 10 mg/kg imiquimod to ferrets elicited emesis with latencies as short as 2'; 12 and 15 mg/kg were optimal doses. Emetic responsiveness was eliminated by complete ablation of the area postrema, but was unaffected by bilateral supradiaphragmatic section of the vagus nerve. This indicates that the emesis is produced by an activation of the chemoreceptor trigger zone B the area postrema. Ferret brain stem slices (450 microm) were preincubated in oxygenated Krebs-Ringer and then mounted in a submerged slice recording chamber. Extracellular recordings of spontaneous and ionophoretically evoked activity of area postrema neurons were obtained for up to 8 h, while the effect of bath-applied imiquimod was determined. Under control conditions, neurons showed a low frequency spontaneous discharge. Introduction of imiquimod (concentration range, 1 x 10(-7) to 5 x 10(-8)M) had no effect on neuronal firing. With ionophoresis of glutamate from an independent micropipette, a brief excitatory response was obtained. We conclude that imiquimod does not directly excite area postrema neurons. It is likely that imiquimod causes synthesis and release of some unknown emetic substance(s), very possibly cytokines.

Aminoquinolines↗

Reciprocal synaptic relationships between angiotensin II-containing neurons and enkephalinergic neurons in the rat area postrema.

A preembedding double immunostaining technique was used to study synaptic relationships between angiotensin-II-like immunoreactive and enkephalin-like immunoreactive neurons in the rat area postrema. The angiotensin-II-like immunoreactive neurons were detected by silver-gold intensification of the DAB reaction results while the enkephalin-like immunoreactive neurons were detected by simple ABC-DAB reaction. The synaptic relationships were reciprocal between the two neurons. Most of the synapses found between these two neurons were the presynaptic enkephalin-like immunoreactive axon terminals that made synapses on the angiotensin-II-like immunoreactive perikarya and dendrites. Both the axo-somatic and axo-dendritic synapses were symmetrical. However, although angiotensin-II-like immunoreactive axon terminals also made synapses on enkephalin-like perikarya and dendrites, the axo-somatic synapses were symmetrical, while the axo-dendritic synapses were asymmetrical. The present results confirm the presence of angiotensin-II-like immunoreactive neurons in the area postrema and suggest that these angiotensinergic neurons in the area postrema may play a role in the regulation of blood pressure via coordinated synaptic interactions with enkephalinergic neurons.

Angiotensin II↗

Forced-choice discrimination of equimolar NaCl and LiCl solutions in rats: effects of ablating the chemosensitive area postrema on acquisition and retention.

The area postrema (AP), a chemosensitive organ located in the fourth ventricle, has been shown to mediate the formation of a lithium-induced conditioned taste avoidance (CTA) in rats. The present experiments examined the role of the AP in the discrimination between two equimolar solutions of sodium chloride (NaCl) and lithium chloride (LiCl). In the first experiment adult male rats were trained to discriminate between equimolar (0.12 M) solutions of NaCl and LiCl in a forced-choice procedure over a 10-day acquisition phase. Subsequently half of the rats (n = 7) received AP lesions (APX) and the other half (n = 7) were given sham lesions (SHAM). In the retention phase all animals were again exposed to the same salt solutions over a 10-day period. Good discrimination (P < 0.001) between the two salt solutions was demonstrated by the end of the acquisition phase and both the APX and SHAM groups exhibited robust retention (P < 0.01) of this discrimination in the second phase. However, when only a LiCl solution was available the APX group ingested significantly more (P < 0.01) than the SHAM rats. No significant group difference emerged when only NaCl was available. In the second experiment rats received ablations of AP or sham lesions and were then trained to discriminate between 0.12 M NaCl and LiCl solutions in a forced-choice procedure over a 10-day period. Both groups exhibited a clear discrimination (P < 0.01) between the two solutions by the end of the acquisition phase. APX rats ingested significantly more LiCl (P < 0.01) than did the SHAM group when this was the only type of fluid available. Again, no such difference was evident when only NaCl was available. These experiments demonstrate that the AP is not necessary for either the acquisition or retention of a discrimination between equimolar solutions of NaCl and LiCl in a forced-choice procedure and that this discrimination is not mediated by a conditioned taste aversion to the LiCl solution.

Animals↗

Mechanisms of vomiting induced by serotonin-3 receptor agonists in the cat: effect of vagotomy, splanchnicectomy or area postrema lesion.

The locations of serotonin-3 (5-HT3) receptors involved in initiating vomiting (emesis) were assessed by cutting visceral afferents or lesioning the area postrema. The 5-HT3 receptor agonists phenylbiguanide (PBG) and 2-methyl-5-HT were shown to induce vomiting and related prodromal signs (e.g., licking, swallowing) in nonoperated cats. Two-methyl-5-HT, but not PBG, also usually produced defecation and sometimes urination. Most studies were conducted using PBG, which induced vomiting in 40/49 (82%) cats at doses of 8.0 mg/kg i.p. or less (thresholds ranged from 2-8 mg/kg, median 5 mg/kg). Latencies to the first episode ranged from 4 to 21 min (median 7.5 min). PBG-induced vomiting was blocked by the 5-HT3 receptor antagonist MDL 72222. Lesions of the area postrema had no apparent effect on vomiting induced by PBG or by electrical stimulation of abdominal vagal afferents. In contrast, the threshold of PBG-induced vomiting was increased by supradiaphragmatic vagotomy and greatly increased by splanchnic nerve section. Thus, abdominal visceral afferents, but not the area postrema, play an important role in mediating vomiting induced by i.p. injection of the 5-HT3 receptor agonist PBG. The mechanisms by which vomiting is induced by PBG as compared to the cancer chemotherapeutic drug cisplatin are discussed.

Animals↗

Neuroactive amino acids in the area postrema. An immunocytochemical investigation in rat with some observations in cat and monkey (Macaca fascicularis).

The localization of five neuroactive amino acids in the rat area postrema was studied by postembedding immunocytochemistry in semithin and ultrathin sections. Antisera to GABA, glycine, glutamate and aspartate produced labelling of cells that were identified as neurons in the electron microscope. GABA-like and glycine-like immunoreactivities occurred in about 20% and 60% of the neurons, respectively, and a minor proportion of the cells displayed both immunoreactivities, suggesting a cellular colocalization of GABA and glycine. Immunoreactivities for glutamate and aspartate were found in a large majority of the neurons, including most of the cells that were positive for GABA and/or glycine. Taurine immunoreactivity was highly concentrated in a few small cells with ultrastructural features typical of microglial cells, and in processes that were probably derived from these. Taurine also appeared to be abundant in cells confined to the perivascular space. The electron microscopic, immunogold analysis of the neuropil revealed numerous nerve terminals that were enriched in GABA or glutamate immunoreactivity, compatible with a transmitter role of these amino acids. Glycine immunolabelling was found preferentially in postsynaptic elements, suggesting that the glycine-containing cells lack locally ramifying axon collaterals, and that they mainly project outside the area postrema. Aspartate immunolabelling was also generally low in axon terminals. This is similar to the situation in several other brain areas and could indicate that the latter amino acid primarily serves metabolic functions in the area postrema.

Animals↗

Fine structure of the ependyma and intercellular junctions in the area postrema of the rat.

Ependymal cells and their junctional complexes in the area postrema of the rat were studied in detail by tracer experiments using horseradish peroxidase (HRP) and colloidal lanthanum and by freeze-etch techniques, in addition to routine electron microscopy. The ependyma of the area postrema is characterized as flattened cells possessing very few cilia, a moderate amount of microvilli, a well-developed Golgi apparatus and rough endoplasmic reticulum. Numerous vesicles or tubular formations with internal dense content were found to accumulate in the basal processes of ependymal cells; the basal process makes contact with the perivascular basal lamina. It is suggested that the dense material in the tubulovesicular formations is synthesized within the ependymal cell and discharged into the perivascular space. The apical junctions between adjacent ependymal cells display very close apposition, with a gap of 2--3 nm, but no fusion of adjacent plasma membranes; they thus represent a transitional form between the zonulae adhaerentes present in the ordinary mural ependyma and the zonulae occludentes in the choroidal epithelium. A direct intercommunication between the ventricular cerebrospinal fluid (CSF) and the blood vascular system indicates that a region exists lacking a blood-ventricular CSF barrier.

Animals↗

Subcellular localization of nitric oxide synthase in the cerebral ventricular system, subfornical organ, area postrema, and blood vessels of the rat brain.

The distribution of neuronal nitric oxide synthase (nNOS) has been studied in the more rostral portion of the lateral ventricle, subfornical organ, area postrema and blood vessels of the rat central nervous system. nNOS was located by means of a specific polyclonal antibody, by using light and electron microscopy. Light microscopy showed immunoreactive varicose nerve fibers and terminal boutons-like structures in the lateral ventricle, positioned in supra- and subependimal areas. The spatial relationships between immunoreactive neuronal processes and the wall of the intracerebral blood vessels were studied. Electron microscopy showed numerous nerve fibers in the wall of the lateral ventricle; many were nNos-immunoreactive and established very close contact with ependymal cells. Immunoreactive neurons and processes were found in the subependymal plate of the ventricular wall, the subfornical organ, the area postrema, and the circularis nucleus of the hypothalamus. In these last three areas, the immunoreactive neurons were found close to the perivascular space of fenestrated and nonfenestrated blood vessels. The nNOS immunoreactivity was localized to the endoplasmic reticulum, cisterns, ribosomes, neurotubules, and in the inner part of the external membrane. In the terminal boutons, the reaction product was found surrounding the vesicle membranes. This distribution showed nNOS as a predominantly membrane-bound protein. The nitrergic nerve fibers present in the wall of the ventricular system might regulate metabolic functions as well as neurotransmission in the subfornical organ, area postrema and circularis nucleus of the hypothalamus.

Animals↗

Catecholamine and serotonin colocalization in projection neurons of the area postrema.

A large population of the rat area postrema (AP) neurons which project to the parabrachial area (PBA) are serotonergic. Many AP neurons contain tyrosine hydroxylase (TH) and are presumed to be noradrenergic. However, it was not previously known whether TH-containing AP neurons also have projections to the PBA. TH-containing and serotonin (5-HT)-containing neurons have wide and overlapping distributions within the AP, and it was therefore possible that TH and 5-HT may be contained within the same AP neurons. In the present study immunohistochemical and retrograde axonal transport techniques were combined to determine whether TH-containing AP neurons project to the PBA and whether TH and 5-HT coexist in AP neurons. Adult male rats were given bilateral injections of the retrograde transport tracer, True blue (TB), into the PBA. After a 4-day survival period, vibratome sections of the caudal brainstem were processed for both TH and 5-HT immunohistochemistry. Examination of the sections revealed that over 25% of the 5-HT and over 30% of the TH-containing AP neurons were retrogradely labelled with TB. More surprising were our findings that many AP neurons displayed both TH and 5-HT immunoreactivity and that almost 40% of these double-labelled cells project to the PBA. Our results indicate that serotonin and noradrenaline coexist in a substantial proportion of the neurons of a major ascending viscerosensory pathway from the AP to the PBA.

Animals↗