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A serotonin-containing pathway from the area postrema to the parabrachial nucleus in the rat.

A combined retrograde tracing-immunofluorescent technique was used to identify the relationships between the cellular population projecting to the parabrachial nucleus and the serotonin-immunoreactive cell population of the area postrema in rats. The retrograde fluorescent tracer True Blue was injected in the parabrachial region and 3 days later the animals were perfused. Serial cryostat sections were processed for serotonin immunofluorescence. Three different groups of labeled cells were identified in the area postrema. First, True Blue-positive cells (up to 250/section) that project to the parabrachial nucleus were observed distributed throughout the area postrema. Second, in the pargyline (a monoamine oxidase inhibitor)-treated animals a large number of serotonergic cells (up to 125/section) was observed distributed throughout the area postrema. There was a tendency to a heavier distribution of serotonin-immunoreactive cells in the dorsal two-thirds of the area postrema. Third, double-labelled cells were also seen. Twenty percent of the True Blue-labelled cells projecting to the parabrachial nucleus were serotonin-immunoreactive. Thirty nine percent of the serotonin-immunoreactive population was retrogradely labelled with True Blue. Thus a new serotonergic pathway from the area postrema to the parabrachial nucleus is described; this pathway may be important in the ascending transmission and modulation of chemical and visceral sensory input.

Animals↗

Role of alpha-adrenergic mechanisms on responses to area postrema stimulation and circulating vasopressin.

This study evaluated the role of catecholamines in the nucleus tractus solitarius (NTS) on the inhibition of renal sympathetic nerve activity (RSNA) due to direct electrical stimulation of the area postrema. In addition, the effects of catecholaminergic mechanisms in the NTS on the ability of circulating arginine vasopressin (AVP) to modulate arterial baroreflex control of RSNA were evaluated. Electrical stimulation of the area postrema (15 microA, 0.3 ms, 5-80 Hz) produced progressive decreases in RSNA. Responses to area postrema activation were not altered by microinjection of the alpha 1-adrenergic antagonist, prazosin, or vehicle bilaterally into the NTS. Microinjection of the alpha 2-antagonist, yohimbine (4 injections of 20-40 nl, 1 ng/nl), unilaterally into the NTS significantly attenuated the RSNA response to area postrema stimulation. Bilateral injection of yohimbine into the NTS abolished the response to area postrema activation (P < 0.05). Baroreflex inhibition of RSNA was significantly greater during infusion of AVP than during infusion of phenylephrine (slopes = -5.18 +/- 0.39 and -2.64 +/- 0.27 for AVP and phenylephrine, respectively). Microinjection of yohimbine bilaterally into the NTS did not alter the slope of baroreflex control of RSNA for phenylephrine but normalized the slope for AVP (-2.85 +/- 0.54) to that of phenylephrine. Data are consistent with the hypothesis that AVP effects on baroreflex inhibition of RSNA and area postrema-mediated inhibition of RSNA require alpha 2-adrenergic signaling within the NTS.

Adrenergic alpha-Antagonists↗

ANG II and baroreflex function in rabbits with CHF and lesions of the area postrema.

Blockade of the angiotensin II (ANG II) type 1 receptor (AT(1)) has been shown to restore baroreflex sensitivity in rats and rabbits with experimental chronic heart failure (CHF). Because the modulation of baroreflex function in response to ANG II is mediated in part by AT(1) receptors located in the area postrema, we hypothesized that lesions of the area postrema would prevent the enhancement in baroreflex function in response to AT(1)-receptor blockade in rabbits with pacing-induced CHF. Experiments were carried out on 24 male New Zealand White rabbits that were divided into sham (n = 12) and lesioned (n = 12) groups further divided into normal and CHF subgroups (n = 6 each). All rabbits were identically instrumented to measure cardiac external dimensions, central venous pressure, arterial pressure, heart rate (HR), and renal sympathetic nerve activity (RSNA). After 3-4 wk of pacing, baroreflex sensitivity (infusions of phenylephrine and nitroprusside) was evaluated before and after intravenous administration of the AT(1)-receptor antagonist L-158,809. Maximum baroreflex sensitivity in nonpaced rabbits was 5.4 +/- 0.7 beats. min(-1). mmHg(-1) and 5.2 +/- 0.5% of maximum/mmHg for HR and RSNA curves, respectively, and was not altered by L-158,809 in either intact or lesioned rabbits. In contrast, L-158,809 enhanced baroreflex sensitivity in intact rabbits with CHF (HR from 1.6 +/- 0.3 to 4.1 +/- 0.7 beats. min(-1). mmHg(-1), P < 0.001; RSNA from 2.3 +/- 0.2 to 4.9 +/- 0.4% of maximum/mmHg, P < 0.001). However, in CHF rabbits with area postrema lesions, L-158,809 failed to enhance baroreflex sensitivity. Interestingly, area postrema lesions did not normalize the baroreflex in CHF rabbits. From these data we conclude that the area postrema mediates the normalization of baroreflex sensitivity after AT(1) blockade in rabbits with CHF but does not modify resting baroreflex function.

Angiotensin II↗

Comparative aspects of the area postrema: fine-structural considerations help to determine its function.

The area postrema is a circumventricular organ of the fourth ventricle of the mammalian brain. Although there are distinct gross anatomical differences in the appearance of this organ between "lower" mammals such as rodents and lagomorphs and "higher" mammals such as carnivores and primates, its fine structure is remarkably similar in all species studied. There are many suggestions in the literature for a specific function for this area of the brain, ranging from its being a chemoreceptive trigger zone for the emetic response to its being a regulatory nucleus for the sleep cycle. The present report describes some comparative studies on the ultrastructure of this organ. This information is discussed in relation to what is known about the neurochemistry of the area postrema and its connections with other brain regions and visceral structures. A suggestion is offered that our current knowledge of the area postrema is consistent with its performing many of its proposed functions in the context of a regulatory ("fine-tuning") center for many autonomic functions.

Animals↗

Interactions between radiation and amphetamine in taste aversion learning and the role of the area postrema in amphetamine-induced conditioned taste aversions.

Three experiments were run to assess the role of the area postrema in taste aversion learning resulting from combined treatment with subthreshold unconditioned stimuli and in the acquisition of an amphetamine-induced taste aversion. In the first experiment, it was shown that combined treatment with subthreshold radiation (15 rad) and subthreshold amphetamine (0.5 mg/kg, IP) resulted in the acquisition of a taste aversion. The second experiment showed that lesions of the area postrema blocked taste aversion learning produced by two subthreshold doses of amphetamine. In the third experiment, which looked at the dose-response curve for amphetamine-induced taste aversion learning in intact rats and rats with area postrema lesions, it was shown that both groups of rats acquired taste aversions following injection of amphetamine, although the rats with lesions showed a less severe aversion than the intact rats. The results are interpreted as indicating that amphetamine-induced taste aversion learning may involve area postrema-mediated mechanisms, particularly at the lower doses, but that an intact area postrema is not a necessary condition for the acquisition of an amphetamine-induced taste aversion.

Amifostine↗

Increase in coronary vascular resistance produced by stimulating neurons in the region of the area postrema of the cat.

Recently, Somberg, in a preliminary report (1983), noted that electrical stimulation of the area postrema causes an increase in coronary vascular resistance. The increase in resistance was mediate by an increase in sympathetic activity as it was counteracted by alpha-adrenergic receptor blockade. The purpose of our study was to confirm the findings of Somberg by using the method of chemical-induced activation of cell bodies in the area postrema. This was done by bilateral topical application of kainic acid to the area postrema of cats while monitoring coronary blood flow, heart rate, arterial blood pressure and the ECG. Topical application of kainic acid produced an increase in coronary vascular resistance (43 +/- 13%, P less than 0.05), S-T segment changes, increases in heart rate (60 +/- 10, P less than 0.05) and arterial blood pressure (88 +/- 20, P less than 0.05) and ventricular tachyarrhythmias. All of these effects were prevented by pretreating animals with the alpha-adrenoceptor blocking agent, phentolamine. These results suggest that chemical excitation of area postrema neurons produces coronary constriction that is mediated by the sympathetic nervous system and alpha-adrenoceptors on coronary vessels.

Adrenergic Fibers↗

Identification of 5-HT3 recognition sites in the ferret area postrema.

The 5-HT3 receptor antagonist [3H]zacopride was used to identify 5-HT3 recognition sites in the ferret area postrema. Specific binding was determined by the inclusion of the 5-HT3 receptor antagonist BRL 43694 in the incubation media, and was shown to be much higher in the area postrema than in other brain regions. The increased binding in the area postrema may reflect either a greater number of binding sites, a higher affinity for such sites, or both. The results indicate that 5-HT3 recognition sites are present within the area postrema and may afford an antiemetic site of action for zacopride and other 5-HT3 receptor antagonists.

Animals↗

Neurons in area postrema mediate vasopressin-induced enhancement of the baroreflex.

Intravenous infusion of arginine vasopressin (AVP) has been shown to enhance baroreflex sensitivity, and this enhancement is dependent on the integrity of the area postrema. However, previous studies did not differentiate a role for cell bodies in the area postrema vs. the dense network of fibers located in and around the lateral and ventral margins of this circumventricular organ. In the present study, baroreflex function was assessed in conscious rabbits by examining heart rate after ramp infusions of phenylephrine (PE) and AVP. The subsequent day, the excitotoxin kainic acid was injected (30 nl initially, with five 15-nl supplemental injections of a 1 ng/nl solution over 1 h) into the area postrema, thus selectively destroying cell bodies. After an 8-day recovery period, baroreflex function was again assessed. The bradycardic response to graded infusion of PE (slope = -2.29 +/- 0.30) was not significantly different after selective lesions of area postrema neurons (slope = -1.88 +/- 0.49). In contrast, the previously enhanced bradycardic response to infusion of AVP (slope = -5.76 +/- 1.02) was significantly attenuated (slope = -2.31 +/- 0.21) to levels similar to that seen with infusion of PE. Thus selective chemical lesions of area postrema neurons block vasopressin-induced enhancement of the baroreflex.

Animals↗

Toxin-induced conditioned changes in taste reactivity and the role of the chemosensitive area postrema.

Conditioned taste avoidances (CTAs) are an important component of behavioral regulation of ingestion. In the laboratory CTAs can be produced by pairing a novel taste stimulus with the physiological feedback produced by a toxin, such as lithium. Such toxins putatively activate a chemosensitive brainstem structure, the area postrema, which ultimately results in the production of a CTA. The present review describes a series of studies which examined conditioned changes in taste reactivity responses (TRRs) when a novel intraoral sucrose taste was paired with the effects of an intraperitoneal (IP) injection of LiCl, and the role of the area postrema in the formation of conditioned palatability shifts. It was first of all necessary to examine the effects of area postrema ablations on TRRs to a range of intraoral sucrose and quinine stimulus intensities. In the first study area postrema lesioned rats exhibited concentration dependent changes in TRRs to these taste stimuli that were very similar to those exhibited by sham lesioned rats. The second study demonstrated that 30 s intraoral infusions of sucrose (0.3 M), presented at 5 or 10 min intervals following an IP injection of LiCl (3.0 meq), resulted in conditioned changes in TRRs. These were characterized by orderly, gradual reductions in ingestive responses and increases in aversive responses. Finally, when area postrema lesioned rats (Study 3) were subjected to this conditioning procedure (brief sucrose presentations paired with the effects of LiCl) no evidence for conditioned or unconditioned changes in TRRs to sucrose were obtained. Lesioned rats injected with LiCl behaved similarly to sham lesioned rats injected with NaCl. These series of studies provide evidence indicating that the chemosensitive area postrema mediates the formation of conditioned palatability shifts induced by treatment with a toxin such as lithium.

Animals↗

Modulation of the arterial baroreceptor reflex by the vasopressin receptor in the area postrema of the hypertensive rats.

The role of arginine8-vasopressin (AVP) in regulation of the baroreceptor reflex in the area postrema was examined in anesthetized hypertensive rats. The sensitivity of the baroreceptor reflex in a one-kidney one clip (1K1C) hypertensive rats was increased in only the initial stage (2 weeks), in association with increase in blood pressure, and then returned to the normal level. This increase in the sensitivity of the baroreceptor reflex in the initial stage was reversed by microinjection of a V1 or V2 antagonist (1 microg) into the area postrema. AVP V2 receptor mRNA was expressed temporarily in the area postrema in this period. These results suggest that vasopressin receptors in the area postrema is important in regulating the sensitivity of the baroreceptor reflex.

Animals↗

Sleep cycles in cats during chronic electrical stimulation of the area postrema and the anterior raphe.

Sleep-waking profiles were obtained from 130 7 hr stimulation-EEG recording sessions in a series of cats bearing chronically implanted stimulating electrodes in the regions of the area postrema and anterior raphe nuclei. The results indicated that: (a) during electrical stimulation of the region of the area postrema with 0.5 or 10 Hz at 1 and 2 mA there were significant increases in the occurrence of the deeper aspects of slow-wave sleep and in REM sleep. These elevations were significant in comparison to nonstimulation baselines and to sleep profiles obtained during stimulation of points located dorsal and anterior to the area postrema. (b) Stimulation of the medial reticular formation including the anterior raphe using the same parameters employed for the area postrema did not alter the occurrence of any stage of sleep. These findings indicate that the region of the area postrema may be more involved in the generation of sleep than the anterior raphe nuclei.

Animals↗

Conditioned taste aversions induced by 1-5-hydroxytryptophan are mediated by the area postrema.

Previous studies have shown that injections of 5-HTP can induce conditioned taste aversions when paired with a novel taste. Adult male albino rats received either lesions of the area postrema or were subjected to a sham lesion procedure. All rats were adjusted to a 23 1/2 hr/day water deprivation schedule and on the conditioning day were given a 0.15% saccharin solution for 1/2 hr. After drinking the saccharin fluid 9 area postrema lesioned and 10 sham lesioned rats were injected i.p. with 25 mg/kg 1-5-hydroxytryptophan. Similarly 10 area postrema lesioned and 6 sham lesioned rats were injected with the vehicle solution. A two-bottle choice test between the saccharin solution and water was given to all animals on the third and fourth days after the conditioning day. The sham lesioned rats injected with the 1-5-hydroxytryptophan exhibited a strong aversion to the saccharin taste whereas the vehicle injected sham lesioned rats showed an equal preference for the two fluids. The difference in group mean saccharin preference ratio was significant (p less than .01). Both area postrema lesioned groups exhibited saccharin preference ratios that were comparable to and not significantly different from the sham lesioned animals injected with the vehicle solution. These results show that an intact area postrema is necessary for induction of conditioned taste aversions with 1-5-hydroxytryptophan.

5-Hydroxytryptophan↗

Involvement of the area postrema in the regulation of sympathetic outflow to the cardiovascular system.

The circumventricular organs of the brain have been implicated in the central regulation of the cardiovascular system. The area postrema, which is the only circumventricular organ in the hindbrain, has received less attention than the others, but recent studies suggest that it may play an important role in the regulation of the cardiovascular system. Studies in rats and rabbits indicate that angiotensin II (Ang II)-dependent hypertension is abolished by lesioning of the area postrema. Additional studies indicate that the hypertension associated with Ang II involves a resetting of the arterial baroreflex to a higher pressure. This upward resetting requires an interaction of neurons in the area postrema with barosensitive neurons terminating in the medial nucleus tractus solitarius (mNTS). Another peptide, arginine vasopressin (AVP), has been shown to enhance the sympathoinhibitory influence of the arterial baroreflex via an action at the area postrema. Studies in rabbits suggest that the sympathoinhibitory response is due to resetting of the baroreflex to a lower pressure. Electrophysiological studies, using an in vitro brain slice preparation, have shown that activation of area postrema neurons projecting to the mNTS alters the responsiveness of mNTS neurons to afferent inputs. It is postulated that alpha-adrenergic mechanisms are involved in these interactions.

Animals↗

Role of the hyperpolarization-activated cation current (Ih) in pacemaker activity in area postrema neurons of rat brain slices.

To clarify the functional properties of the hyperpolarization-activated cation current (Ih) as a pacemaker current in area postrema neurons, whole-cell recordings were made in visually identified cells in rat brain slices. The activation of Ih was identified in approximately 62 % of area postrema neurons tested. The cells displaying Ih showed a depolarizing "sag" in responses to hyperpolarizing current injection in current-clamp mode. The reversal potential for the Ih was -36 mV, and this was shown to depend on the external concentration of Na+ and K+ ions. Extracellular Cs+ ions (2 mM) and ZD7288 (100 microM), a potent selective Ih channel antagonist, blocked Ih and induced a membrane potential hyperpolarization, suggesting the sustained activation of Ih near the resting potential and a contribution from Ih to membrane potential maintenance at more depolarized levels. In contrast, extracellular Ba2+ ions caused a depolarization of the membrane potential, suggesting the blockade of inward rectifier K+ currents. ZD7288 decreased the spontaneous discharge rate by prolonging the slow depolarization between two spikes, with minimal effect on the amplitude of the afterhyperpolarization or action potential waveforms. Ih stabilized the latency of rebound action potentials. Ih was weakly activated by external 8-bromoadenosine 3',5' cyclic monophosphate (1 mM) or forskolin (50-100 microM), indicating that the Ih channel subtypes in area postrema cells could be modulated by intracellular cAMP. Our findings indicate that Ih contributes to the subthreshold membrane and firing properties of rat area postrema neurons and may regulate their resting membrane potential and firing patterns.

Action Potentials↗

Localization of tyrosine hydroxylase in neuronal targets and efferents of the area postrema in the nucleus tractus solitarii of the rat.

Catecholamines in the nucleus tractus solitarii (NTS) have been implicated in autonomic responses to circulating hormones that act on neurons in the area postrema, the most caudal circumventricular organ in brain. We combined immunoperoxidase labeling of the anterograde tracer, Phaseolus vulgaris leucoagglutinin (PHAL) with immunogold-silver labeling of tyrosine hydroxylase to determine whether this enzymatic marker for catecholamines was present in efferents from the area postrema or their targets in the rat NTS. At survival periods of 10-12 days after PHAL injections into the area postrema, light microscopy revealed numerous varicose processes containing peroxidase reaction product for PHAL in the dorsomedial, medial, and commissural NTS. Some of these labeled processes were located near neuronal perikarya and processes containing immunogold-silver intensified reaction product for tyrosine hydroxylase. Electron microscopy of the commissural and dorsomedial NTS established that the majority of the labeling for PHAL was in axon terminals, whereas immunogold labeling for tyrosine hydroxylase was mainly in soma and dendrites. Only 3 out of 579 PHAL-labeled terminals also contained detectable tyrosine hydroxylase immunoreactivity. Fifty-eight percent (335/579) of the PHAL-labeled terminals formed synapses with recognized symmetric junctions, whereas the remainder lacked synaptic specializations within the examined series of serial sections. Of those PHAL terminals forming recognized symmetric junctions, 22% were on tyrosine hydroxylase-immunoreactive dendrites, 74% on unlabeled dendrites and 4% on unlabeled axon terminals. From a total of 1,250 observed contacts on tyrosine hydroxylase labeled dendrites, 88 (7%) contained PHAL, 9 (< 1%) contained TH, and 1,180 (93%) lacked detectable immunoreactivity and formed primarily symmetric synapses. We conclude that a few catecholamine, but mainly noncatecholamine efferents from the area postrema provide a monosynaptic, and most likely inhibitory input to target neurons both with and without tyrosine hydroxylase immunoreactivity in the dorsomedial and commissural NTS. Synapses between the efferent terminals from the area postrema and tyrosine hydroxylase labeled and unlabeled dendrites as well as unlabeled axons in these specific subnuclei of the NTS suggest multiple sites for modulation of gastric and cardiovascular reflexes in response to circulating peptides.

Animals↗

Fine structure of the ependymal cells in the area postrema of the domestic fowl.

The ultrastructure of the ependymal cells in the area postrema of the domestic fowl was studied by scanning and transmission electron microscopy. The ependymal surface of the area postrema is covered with many furrows and ridges. These ridges consist of ependymal cells aggregated in a fan-like shape. The ependymal cell lacks clustered cilia, microvilli are few, and a long basal process extends through the parenchymal layer of the area postrema. Within the cytoplasm as well as in the basal process, a spherical body with a diameter ranging from 1.5 to 2 micron is occasionally observed.

Animals↗

Membrane-bound and solubilized brain 5HT3 receptors: improved radioligand binding assays using bovine area postrema or rat cortex and the radioligands 3H-GR65630, 3H-BRL43694, and 3H-LY278584.

Bovine area postrema tissue was used as a convenient source of tissue for studies of brain 5HT3 receptors. 5HT3 receptor density was determined to be 97 +/- 5 fmol/mg and 124 +/- 10 fmol/mg of protein using the commercially available 5HT3 radioligands, 3H-GR65630 and 3H-BRL43694, respectively. The equilibrium dissociation constants (KD) for 3H-GR65630 and 3H-BRL43694 were 0.5 +/- 0.1 nM and 1.7 +/- 0.3 nM, respectively. The affinities of a series of drugs for the 5HT3 receptor using the two radioligands were essentially identical; the Ki values and order of affinities of agonists and antagonists were very similar to data published in studies on radiolabeling of 5HT3 receptors in rat brain. 3H-LY278584 also labeled 5HT3 receptors in bovine area postrema homogenates with a KD of 3.1 +/- 0.1 nM and a Bmax of 84 +/- 6 fmol/mg. In rat cortical homogenates, 3H-LY278584 produced the most reliable specific signal of 72%, with a KD of 2.6 +/- 0.3 nM and a Bmax value of 10.5 +/- 1 fmol/mg. At 1 nM, 3H-GR65630 or 3H-BRL43694 specific binding represented 28 and 50% of total radioligand binding, respectively. These data in bovine and rat brain tissues indicate that bovine area postrema can be used with 3H-GR65630, 3H-BRL43694, or 3H-LY278584 for drug screening or molecular investigations of the brain 5HT3 receptor: only 3H-LY278584 can be used for studies on the regulation and/or the molecular properties of 5HT3 receptors in rat cortical homogenates. 5HT3 receptors were solubilized from bovine area postrema and characterized using 3H-GR65630.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Two distinct types of transient outward currents in area postrema neurons in rat brain slices.

We investigated the electrophysiological properties of the area postrema neurons in acutely prepared rat brain slices using the whole-cell patch-clamp technique. Two different types of transient outward potassium current (I(to)), fast and slow, were found in the area postrema. Both the decay time constant and rise time were significantly faster in the fast I(to) than in the slow I(to). Both current-clamp and voltage-clamp recordings revealed that the activation of fast and slow I(to) contributes to generation of the different spiking patterns, late spiking and interrupted spiking, respectively. The activation and inactivation of both I(to) were strongly voltage-dependent. Curve fitting by the Boltzmann equation revealed no significant difference in the activation and inactivation curves for each I(to) except that the slope factor of inactivation was larger for fast I(to). Both I(to) were suppressed dose-dependently by application of 4-aminopyridine. Each spiking pattern was enhanced when cells were held at a more hyperpolarized membrane potential, i.e. a longer latency of the first spike or longer interspike interval between the first and second spikes. The voltage-dependent modulation of the spiking pattern was consistent with the voltage-dependent activation of I(to). The present study shows significant subdivisions of the area postrema neurons distinguished by a difference in the kinetics of I(to) and spiking patterns. We discuss the role of I(to) as the ionic current underlying neuronal excitability.

4-Aminopyridine↗