Circulating endothelin influences area postrema neurons

The recently described endothelium-derived constricting factor endothelin (ET) is a 21 amino acid peptide which is the most potent endogenous vasoconstrictor yet described. Binding sites for this peptide have been demonstrated within the circumventricular structures of the brain. One of these structures, the area postrema (AP), has been implicated in central cardiovascular control mechanisms. We have recently demonstrated that microinjection of ET into this structure results in dose-dependent changes in mean arterial blood pressure. The present studies were undertaken to test the hypothesis that ET elicits these effects as a result of influences on the activity of AP neurons. Using extracellular single unit recording techniques we have examined the effects of systemic administration of ET on the activity of AP neurons. A total of 60 AP neurons were tested for effects of ET (0.1-10.0 pmol) of which the spontaneous activity of 32 showed rapid (modified frequency of action potentials in the 60s following ET), reversible (return to baseline activity within 10 m) responses to this peptide. The initial response of the majority (84%) of AP neurons influenced by ET was excitatory, while a smaller proportion of AP neurons were inhibited (16%) by systemic administration of this peptide. We have also examined whether such excitatory effects were specific to AP neurons by comparing the above response characteristics to those observed in neurons in the adjacent commissural NTS. Such recordings demonstrated predominantly inhibitory (84% of influenced cells) responses of this group of NTS neurons to ET. While these findings demonstrate specific excitatory effects of systemic ET on the activity of AP neurons they also suggest a potential role for this peptide in controlling the activity of NTS neurons. These studies provide evidence that circulating ET influences AP neuronal function, although they offer no definitive information as to the specific site of action.     

The microvascular pattern and perivascular linings of the area postrema

Summary The microvasculature and perivascular linings of the area postrema (A.P.) were studied electron microscopically with the ultrathin section and freeze-etching techniques. Special attention was given to the intercellular contacts of the different cellular entities. Two types of microvascular segments were identified. The endothelium of these vascular segments reveals fenestrations and a high pinocytotic activity. There are no significant differences in the frequency and distribution of the endothelial openings between both types of capillaries. The endothelium of the blood vessels, however, is joined by different types of tight junctions. Focal tight junctions occur between pericytes and the endothelium, and between leptomeningeal cellular elements in the perivascular space. The cell membrane of the perivascular glia shows intramembrane particles which are either distributed at random or organized in the form of membrane-associated orthogonal particle complexes (MOPC, Dermietzel, 1974). The significance of these findings is discussed with respect to the modified blood-brain barrier mechanism in the A.P.Supported by a grant of the Deutsche Forschungsgemeinschaft (SFB 114 Bionach) to R. DermietzelThe authors are indebted to Mrs. D. Schünke and Mr. R. Eichner for technical assistance and Mr. A. Stapper for preparing the diagram     

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.     

The fine structure of the area postrema of the sheep

The ultrastructural features of the area postrema (AP) were investigated in the suckling lamb, weaned lamb and adult sheep. No morphological differences were observed between lambs and sheep. Unciliated ependymal cells, linked by zonulae adherentes-type junctions and gap junctions, cover the AP ventricular surface. Clusters of pyriform neurons, glial cells, and axons are present in the parenchyma. The blood vessels are surrounded by wide perivascular spaces, which present an inner and outer basal lamina. The capillaries are of the fenestrated type. Perivascular glial cells rest on the outer basal lamina of the perivascular space and form a continuous ensheathment with their cell bodies or with flattened interdigitating processes. Along adjacent perivascular glial processes gap junctions are present. From our ultrastructural observations it appears that the overall cellular morphology of AP of the sheep does not differ substantially from that of monogastric mammals.     

Fine structure of the area postrema of the rabbit brain

Summary The area postrema of the rabbit, which was perfused with glutaraldehyde and postfixed in osmium tetroxide, was observed under the electron microscope. This area showed neuronal and neuroglial structures similar to those of ordinary cerebral tissue, except for rich blood capillaries, which were surrounded by conspicuous perivascular spaces. Parenchymal cells included a moderate number of small neurons and large numbers of specific astrocyte-like cells. The neuropil consisted of a small number of thin myelinated and many non-myelinated nerve fibers of varying calibers, axo-dendritic synapses, and neuroglial cell processes, leaving no spaces between them. The axons and synaptic terminals contained moderate amounts of granular vesicles, which were similar in size to those found in the hypothalamus and were supposed to contain catecholamine. Glycogen paticles were demonstrated mainly in the cytoplasm of the astrocyte-like cells.     

Adrenomedullin influences dissociated rat area postrema neurons

The area postrema (AP) is one of a specialized group of central nervous system (CNS) structures devoid of a significant blood-brain barrier (BBB), collectively known as the circumventricular organs (CVO). While peptides are normally excluded from access to most regions of the CNS, the AP contains neurons with a high density of receptors for many circulating peptides, very likely including those for adrenomedullin (AM). In this study, whole-cell patch-clamp recordings were obtained from 114 dissociated rat AP neurons. The mean resting membrane potential (RMP) of these neurons (n=79) was -54.3+/-0.8 mV, the mean input resistance (IR) was 3.1+/-0.2 GOmega and the spike amplitude of neurons included in this study was always greater than 90 mV. Current-clamp studies showed that bath application of AM depolarized 39.2% (31 of 79) and hyperpolarized 45.6% (36 of 79) of neurons tested. Both effects were found to be concentration dependent from 10(-12) to 10(-7) M. These data support the idea that specific populations of CNS neurons within the AP are directly influenced by AM and support the concept that AM may act at AP to influence central autonomic control. We also examined the roles of specific ion channels in regulating the AM-induced excitability of AP neurons through voltage-clamp studies. These experiments suggest potential actions of AM in modulating voltage gated calcium channels, effects which have the additional consequence of inhibiting calcium activated potassium conductances (I(K(Ca))). These data demonstrate direct effects of AM on dissociated AP neurons and identify ion channels, the modulation of which, may underlie these effects.     

Excitation of neurons in the canine area postrema by prostaglandins

The effects of prostaglandins on electrical activity of neurons in the canine area postrema were studied using the techniques of extracellular recording with iontophoresis. Excitatory responses were obtained upon application of prostaglandins A1, B1, B2, E1, F1 alpha, and F2 alpha in between 24 and 50% of the cells studied. The excitation was very similar in pattern to that observed to apomorphine, biogenic amines, and several neuropeptides in that it had a relatively long latency, low maimal frequency, and prolonged duration. Since the area postrema is known to play a central receptive role in initiating emesis to circulating toxins, these results suggest that prostaglandins may play a role in the initiation of some forms of emesis.     

Chronic sodium depletion suppresses the area postrema pressor pathway

Sodium depletion in dogs is known to affect both the renin-angiotensin as well as the sympathetic nervous system. The effect of this dietary regime upon the area postrema pressor pathway, as evaluated by the cardiovascular responses to centrally acting angiotensin II, has not been determined previously. With this in mind, male mongrel dogs were maintained on either a normal or a sodium restricted diet supplemented with furosemide and dose-response curves for intravertebral and intravenous angiotensin II (range: 1-20 ng/kg/min) were obtained. Sodium depletion results in not only a blunted intravenous pressor response to angiotensin II but also the abolition of the centrally mediated pressor responses mediated by the area postrema. Because accumulating evidence indicates that in sodium depleted dogs sympathetic nerve activity is reduced while central noradrenergic inhibitory activity is increased the reduced effects of angiotensin II upon the central sympathetically mediated pressor response may in part be related to decreases in sympathetic nerve activity.     

Cholecystokinin-octapeptide like immunoreactivity in the area postrema of the rat and cat

The distribution of cholecystokinin-8 (CCK-8)-like immunoreactivity in the area postrema of the rat and cat was visualized using the peroxidase, antiperoxidase technique. In the rat the greatest amount of immunostaining occurred in peripheral regions of the area postrema at intermediate and rostral levels. Caudally, scattered immunoreactivity predominated. After colchicine treatment, numerous immunoreactive somata were observed throughout the area postrema. The cat area postrema had a different and more complex pattern of immunostaining than the rat. Moderate to dense accumulations of immunostaining occurred in the ventromedial region of the area postrema bordering the solitary tract and dorsal vagal nuclei. The central region of the area postrema possessed scattered amounts of immunoreactivity at rostral levels. Following colchicine treatment, no visible CCK-8-like immunoreactive cell bodies were observed in the cat area postrema. Results of the present investigation provide morphological evidence for the role of CCK-8 in cardiovascular regulation and satiety. The difference in the distribution of CCK-8 in the rat and cat suggest a possible role in the emetic reflex.     

Ultrastructure of the area postrema of the monkey, Macaca fascicularis

The area postrema of the monkey, Macaca fascicularis, were a pair of oval organs at the caudal end of the floor of fourth ventricle. Their ependymal lining was covered by well-developed microvilli with occasional overlying supraependymal cells. Two types of lining cells were present: pyramidad- and flattened cells. The pyramidal cell showed a long extending basal process resting on the underlying blood vessels. In transmission electron microscopy, the organ showed numerous fenestrated sinusoids characterized by a distinct perivascular space containing mast cells, macrophages and collagen fibrils. The parenchyma of the organ was composed of neurons and glial elements. Only one type of neuron ranging from 9.5 to 15 microns could be distinguished. The neurons contained an indented nucleus surrounded by organelle rich cytoplasm. The soma of the neuron was enclosed by glial element resembling astrocyte. The glial processes terminated on the blood vessel where they were "tunnelled" by a variable number of nerve fibres some of which gained a direct access to the external basal lamina of the perivascular space. Synapses in the neuropil predominantly of the axodendritic variety were observed. Axon terminals containing round agranular vesicles were seen to make synaptic contacts with the neuronal soma. No structural changes were observed in the area postrema following bilateral cervical vagotomy. However, degenerating axon terminals were observed in the subpostremal zone 7, 14 and 21 days after vagotomy suggesting a direct afferent projection into this region.     

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.