Area Postrema Cells Research Articles

The area postrema does not modulate the long-term salt sensitivity of arterial pressure.

The hindbrain circumventricular organ, the area postrema (AP), receives multiple signals linked to body fluid homeostasis. In addition to baroreceptor input, AP cells contain receptors for ANG II, vasopressin, and atrial natriuretic peptide. Hence, it has been proposed that the AP is critical in long-term adjustments in sympathetic outflow in response to changes in dietary NaCl. The present study was designed to test the hypothesis that long-term control of arterial pressure over a range of dietary NaCl requires an intact AP. Male Sprague-Dawley rats were randomly selected for lesion of the AP (APx) or sham lesion. Three months later, rats were instrumented with radiotelemetry transmitters for continuous monitoring of mean arterial pressure (MAP) and heart rate and were placed in individual metabolic cages. Rats were given 1 wk postoperative recovery. The dietary salt protocol consisted of a 7-day period of 1.0% NaCl (control), 14 days of 4.0% NaCl (high), 7 days of 1.0% NaCl, and finally 14 days of 0.1% NaCl (low). The results are reported as the average arterial pressure observed on the last day of the given dietary salt period: APx (n = 7) 114 +/- 2 (1.0%), 110 +/- 3 (4.0%), 110 +/- 3 (1.0%), and 114 +/- 4 (0.1%) mmHg; sham (n = 6) 115 +/- 2 (1.0%), 114 +/- 3 (4.0%), 111 +/- 3 (1. 0%), and 113 +/- 2 (0.1%) mmHg. Neither group of rats demonstrated significant changes in MAP throughout the entire dietary salt protocol. Furthermore, no significant differences in MAP were detected between groups throughout the protocol. All lesions were histologically verified. These results suggest that the area postrema plays no role in long-term control of arterial pressure during chronic changes in dietary salt.

Indomethacin attenuates oxytocin and hypothalamic-pituitary-adrenal axis responses to systemic interleukin-1 beta.

Systemic administration of the cytokine IL-1 beta produces a significant release of ACTH into the plasma and activation of hypothalamic oxytocin (OT) and corticotropin releasing factor (CRF) cells. However, the mechanism(s) by which systemic IL-1 beta induces these responses is not clear. In the present study, we have investigated the proposal that catecholamine cells of the ventrolateral medulla (VLM) and nucleus of the solitary tract (NTS) can relay circulating IL-1 signals via a prostaglandin-dependent mechanism to effect the HPA axis responses in the rat. Intra-arterial administration of IL-1 beta (1 pg/kg) to otherwise untreated animals produced a prominent release of ACTH into the plasma, substantial c-fos expression in paraventricular medial parvocellular (mPVN) corticotropin releasing factor (CRF) cells, supraoptic (SON) and paraventricular nucleus (PVN) OT cells, area postrema cells, NTS and VLM catecholamine cells and cells of the central amygdala. Pretreatment with the prostaglandin synthesis inhibitor, indomethacin (10 mg/kg body weight ia) 15 min before IL-1 beta administration (1 pg/kg ia) significantly reduced plasma ACTH release and c-fos expression in PVN and SON OT cells and MPVN CRF cells, in addition, the area postrema, A1 and C1 catecholamine cell groups of the VLM and A2 and C2 catecholamine cell groups of the NTS, all exhibited concomitant reductions in c-fos expression. Conversely indomethacin administration did not alter the IL1 beta-induced expression of c-fos in the central amygdala. These data suggest that central pathways involved in the IL-1 beta-induced activation of the HPA axis and OT cells are, at least in part, dependent upon prostaglandin synthesis. It is proposed that neurons in the area postrema, NTS and VLM might mediate this IL-1 beta-induced activation of hypothalamic CRF and OT cells and release of ACTH into the plasma.

Catecholaminergic systems in the zebrafish. III. organization and projection pattern of medullary dopaminergic and noradrenergic neurons

To characterize the catecholaminergic systems in the zebrafish medulla, immunocytological studies were performed by using antibodies directed against tyrosine hydroxylase and dopamine-beta-hydroxylase. Catecholaminergic neurons could be categorized into three populations based on location, dendritic morphology, axonal projection pattern, and targets: an interfascicular group, a vagal area group, and an area postrema group. All groups contained both dopaminergic and noradrenergic neurons. Interfascicular neurons formed a loose longitudinal column of approximately 16-20 multipolar neurons on each side of the medulla, whose rostrocaudal extension coincided roughly with the vagal lobe. These neurons were relatively large and had dendrites that arborized throughout the reticular formation and in the vagal lobe. They also contributed axonal processes to the longitudinal catecholamine bundle. Neurons associated with structures in the vagal area were mostly dopaminergic. Some cells had a short, thin apical process that arborized into a dense plexus near the ventricular surface, and all cells had a basal dendrite that divided into two main branches: one extended caudally to terminate in the commissural nucleus of Cajal and among the postobecular catecholaminergic cell group; the other extended laterally and joined the longitudinal catecholamine bundle. The caudal extent of this cell group reached the medullospinal junction. The area postrema cell group consisted of densely packed, bipolar neurons. One process of these neurons contacted the ventricular surface in the area postrema, and one terminated in the commissural nucleus of Cajal. Collaterals from the latter innervated the superficial laminae of the vagal lobe and joined the longitudinal catecholamine bundle. The longitudinal catecholamine bundle ascended through the medulla ventral to the secondary gustatory tract. Whether some fibers extended more rostrally is not known. The majority of the terminal fields of medullary catecholaminergic neurons appeared to be restricted to the medulla and were strongly associated with sensory systems. With the exception of some cells in the vagal area, catecholamine-containing neurons in the zebrafish medulla were not obviously homologous to those in the mammalian brainstem.

Angiotensin II and glutamate influence area postrema neurons in rat brain slices

The area postrema (AP) has been repeatedly implicated in cardiovascular regulation. Microinjection and single unit recording studies in vivo have suggested specific actions for angiotensin II (ANG) and glutamate (GLU) in controlling the excitability of AP neurons. The present study was therefore designed to examine the responsiveness of AP neurons to bath administration of these substances. Of the 133 AP neurons tested with ANG (10−8–10−6 M) 40% were excited, 13% inhibited and the remainder unresponsive. The excitatory effects of ANG on AP neurons were dose-dependent. Following blockade of synaptic transmission with a low calcium high magnesium solution excitatory responses were maintained in 12 of 15 cells tested. Pretreatment of slices with the AT1 receptor antagonist losartan blocked the excitatory effects of ANG in all cells (55) tested. The effects of GLU on AP neurons were also examined. Of the 71 AP cells tested, 40% were excited, 10% inhibited, 8% showed excitatory responses followed by periods of inhibition while the remaining cells were unaffected. Excitatory effects of GLU were maintained in all AP neurons (77) tested during perfusion with low calcium, high magnesium solutions. Similar responses to NMDA were observed in four of four cells tested, suggesting these GLU actions are mediated through NMDA receptors. These data demonstrate direct excitatory actions of ANG and GLU on AP neurons which are likely mediated through the AT1 and NMDA receptors, respectively.

Angiotensin II and glutamate influence area postrema neurons in rat brain slices

The area postrema (AP) has been repeatedly implicated in cardiovascular regulation. Microinjection and single unit recording studies in vivo have suggested specific actions for angiotensin II (ANG) and glutamate (GLU) in controlling the excitability of AP neurons. The present study was therefore designed to examine the responsiveness of AP neurons to bath administration of these substances. Of the 133 AP neurons tested with ANG (10 −8–10 −6 M) 40% were excited, 13% inhibited and the remainder unresponsive. The excitatory effects of ANG on AP neurons were dose-dependent. Following blockade of synaptic transmission with a low calcium high magnesium solution excitatory responses were maintained in 12 of 15 cells tested. Pretreatment of slices with the AT 1 receptor antagonist losartan blocked the excitatory effects of ANG in all cells ( 5 5 ) tested. The effects of GLU on AP neurons were also examined. Of the 71 AP cells tested, 40% were excited, 10% inhibited, 8% showed excitatory responses followed by periods of inhibition while the remaining cells were unaffected. Excitatory effects of GLU were maintained in all AP neurons ( 7 7 ) tested during perfusion with low calcium, high magnesium solutions. Similar responses to NMDA were observed in four of four cells tested, suggesting these GLU actions are mediated through NMDA receptors. These data demonstrate direct excitatory actions of ANG and GLU on AP neurons which are likely mediated through the AT 1 and NMDA receptors, respectively.

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)

Immunohistochemical analysis of rat arthritic synovia by means of lymphocyte and macrophage markers

Amylin, which is co-secreted together with insulin by pancreatic beta cells, is considered to be an important peptide hormone involved in the control of feeding behavior and energy homeostasis. Although the area postrema has been implicated to be a primary target of amylin, there are no studies of the mechanisms by which amylin may alter the excitability of area postrema neurons. To investigate the mechanism for amylinergic modulation of neuronal excitability, we performed perforated patch-clamp recordings from area postrema neurons in rat brainstem slices. Amylin-induced changes in excitatory responses, such as increases in the frequency of mEPSCs (miniature excitatory postsynaptic currents) and changes in the amplitude distribution of mEPSCs, were found in cells not displaying the hyperpolarization-activated cation current (Ih). Area postrema cells displaying Ih did not respond to amylin application. Inhibitory responses to amylin were never encountered. Bath application of CNQX (AMPA type glutamate receptor antagonist) abolished the effects of amylin. Depolarization of cells during amylin application was sufficient at 1 μM to cause action potential discharge by responding cells. We conclude that amylin receptors are located mostly on presynaptic glutamatergic terminals connecting to the area postrema neurons not displaying Ih and amylin concentrations can increase glutamate release enough to cause cell firing. Modulation of amylinergic activity may offer a novel target to influence food intake and obesity.

Anti-inflammatory effect of Amylin 8–37 fragment in some models of inflammation

Amylin, which is co-secreted together with insulin by pancreatic beta cells, is considered to be an important peptide hormone involved in the control of feeding behavior and energy homeostasis. Although the area postrema has been implicated to be a primary target of amylin, there are no studies of the mechanisms by which amylin may alter the excitability of area postrema neurons. To investigate the mechanism for amylinergic modulation of neuronal excitability, we performed perforated patch-clamp recordings from area postrema neurons in rat brainstem slices. Amylin-induced changes in excitatory responses, such as increases in the frequency of mEPSCs (miniature excitatory postsynaptic currents) and changes in the amplitude distribution of mEPSCs, were found in cells not displaying the hyperpolarization-activated cation current (Ih). Area postrema cells displaying Ih did not respond to amylin application. Inhibitory responses to amylin were never encountered. Bath application of CNQX (AMPA type glutamate receptor antagonist) abolished the effects of amylin. Depolarization of cells during amylin application was sufficient at 1 μM to cause action potential discharge by responding cells. We conclude that amylin receptors are located mostly on presynaptic glutamatergic terminals connecting to the area postrema neurons not displaying Ih and amylin concentrations can increase glutamate release enough to cause cell firing. Modulation of amylinergic activity may offer a novel target to influence food intake and obesity.

Circulating vasopressin influences area postrema neurons

Extracellular single-unit recordings were obtained from 107 area postrema and 74 nucleus tractus solitarius neurons in sodium pentobarbitol anaesthetized rats. Systemic administration of vasopressin (1–10 ng) decreased the firing frequency of 45.8% of area postrema neurons and 58.1% of nucleus tractus solitarius neurons tested while the firing frequency of 38.3% of area postrema neurons and 21.6% of nucleus tractus solitarius neurons was increased by this peptide. To determine whether these neurons were specifically influcenced by vasopressin or the accompanying pressor response, the effects of α-adrenergic agonists on neuronal activity were also determined. Cells that responded similarly to vasopressin and the change in blood pressure elicited by a-adrenergic agonists were classified as “blood pressure-sensitive”, whereas those neurons that responded differently to both agents were classified as “vasopressin-sensitive” neurons. The majority (85.2%) of area postrema cells that decreased firing frequency in response to vasopressin were determined to be “vasopressin-sensitive”, while 68.8% of area postrema neurons responding to vasopressin with increases in firing frequency were classified as “blood pressure-sensitive”. In contrast, 78.6% of nucleus tractus solitarius neurons that decreased firing frequency in response to vasopressin and 55.5% of those that increased firing frequency were classified as “blood pressure-sensitive” neurons. To determine whether the actions of vasopressin in the area postrema were mediated by V 1 receptors the effect of vasopressin after V 1 receptor blockade was examined in seven “vasopressin-sensitive” area postrema neurons. All seven neurons tested showed no response to vasopressin after such V 1 receptor blockade. These data suggest that there exists a population of area postrema neurons specifically responsive to circulating vasopressin as a result of actions of this peptide at V 1 receptors. They also implicate these neurons in the physiological mechanisms through which circulating vasopressin acts in the area postrema to influence baroreceptor reflex sensitivity.

Electrophysiological characterization of reciprocal connections between the parabrachial nucleus and the area postrema in the rat

Neuroanatomical studies have demonstrated reciprocal connections between the parabrachial nucleus (PBN) and both the area postrema (AP) and the nucleus tractus solitarius (NTS). To functionally characterize these projections, antidromic identification of AP and NTS neurons projecting to the PBN was attempted. Orthodromic influences on these cells, resulting from PBN stimulation, were also examined. Four percent of AP neurons tested (n = 74) were antidromically identified as projecting to the PBN [latency (L) = 26 ± 4 msec, threshold current (T) = 79 ± 11 μA]. Parabrachial stimulation orthodromically influenced 24% of AP cells. Equal numbers of these neurons (12%) were excited [L = 25 ± 9 msec, duration (D) = 29 ± 14 msec] and inhibited (L = 28 ± 8 msec, D = 107 ± 40 msec). Of 46 NTS neurons tested, 11% were antidromically identified as projecting to the PBN (L = 12 ± 4 msec, T = 61 ± 18 μA), while orthodromic influences were seen in 41 % of these neurons. Initial responses of 30% of the cells were excitatory (L = 34 ± 14 msec, D = 63 ± 24 msec), PBN stimulation inhibited the remaining 11% of NTS neurons (L = 30 ± 10 msec, D= 108 ±32 msec). These findings suggest that a functional heterogeneity exists in the PBN efferents to the AP and NTS. However, the small proportion of antidromically identified AP and NTS efferents to the PBN disagrees with neuroanatomical studies suggesting a denser projection.

Electrophysiological evidence that systemic angiotensin influences rat area postrema neurons

Extracellular single-unit recordings from neurons in the area postrema (AP) and the nucleus tractus solitarius (NTS) in anesthetized male rats demonstrated that most cells in these regions have spontaneous activities of 5 Hz or less. Systemic angiotensin (ANG II) (50-500 ng) enhanced the activity of 55% of AP cells tested (n = 76), whereas 53% of tested NTS neurons (n = 62) were inhibited by ANG II. To determine whether these neurons were influenced specifically by circulating ANG II or by the accompanying increase in mean arterial blood pressure (BP), the effects of adrenergic agonists given intravenously on ANG II influenced neurons were also examined. Subsequently two cell types were characterized: cells responding to iv ANG II but not to the adrenergic agonist ("ANG II sensitive") and cells responding in a similar way to both agents ("BP sensitive"). Most ANG II-responsive neurons in the AP (53.5%) and the NTS (65%) were determined to be BP sensitive. These data demonstrate that ANG II influences the activity of AP neurons. In addition, there exists a second population of AP neurons apparently responsive to perturbations of the cardiovascular system. These studies further emphasize the potential roles of the AP in the regulation of body fluid balance.

Central neurotoxic effects of guanethidine: altered serotonin and enkephalin neurons within the area postrema

This study reveals that the 5-HT and ankephalin cell body population of the area postrema (AP) is dramatically reduced via exposure to the peripherally circulating neurotoxin guanethidine. Efforts to restore cell body numbers to control levels, with colchicine and monoamine oxidase inhibitors, subsequent to guanethidine treatment are ineffective. However, 5-HT and enkephalin immunostaining of surrounding bulbar nuclei appears normal, implying that the neurotoxic effect of guanethidine is restricted to the AP. In addition, γ-aminobutyric acid and neurotensin immunostaining within the AP appears normal, subsequent to treatment, suggesting that the neurotoxic effect is restricted to specific AP cell populations.