Morphological, histological and immunohistochemical study of the area postrema in the dog.

Circumventricular organs are specialized brain structures that are located mainly at the midsagittal line, around the third and fourth ventricles, often protruding into the lumen. They are positioned at the interface between the neuroparenchyma and the ventricular system of the brain. These highly vascularized nervous tissue structures differ from the brain parenchyma, as they lack a blood-brain barrier. Circumventricular organs have specialized sensory and secretory functions. It is essential for any pathologist who evaluates brain sections to have a solid knowledge of microscopic neuroanatomy and to recognize these numerous specialized structures within the nervous system as normal and not mistake them for pathological changes. The purpose of this study was to provide, for the first time, a detailed and complete histological description of the healthy canine area postrema and to determine its resemblance to that of other mammalian species. Anatomical dissections with routine histological and immunohistochemical techniques were carried out on ten canine brains. The cellular composition of area postrema proved to be largely comparable to that of other mammal species.

Effects of +Gz Loads on Structural Organization of Central Autonomic Nuclei.

Structural alterations in the central autonomic nuclei (dorsal vagal complex and intermediolateral nucleus) of the centrifuged random-bred male rats subjected to +Gz loads were examined. Acute exposure to gravitational loads predominantly produced the reactive changes in these nuclei, while persistently repeated regular loads resulted in cumulation of the destructive alterations. The structural perturbations in the central autonomic nuclei can disturb the autonomic regulation of physiological functions. The character of such disturbances is partially determined by the peculiarities in structural organization of these nuclei.

Changes in pericytic expression of NG2 and PDGFRB and vascular permeability in the sensory circumventricular organs of adult mouse by osmotic stimulation.

The blood-brain barrier (BBB) is a barrier that prevents free access of blood-derived substances to the brain through the tight junctions and maintains a specialized brain environment. Circumventricular organs (CVOs) lack the typical BBB. The fenestrated vasculature of the sensory CVOs, including the organum vasculosum of the lamina terminalis (OVLT), subfornical organ (SFO) and area postrema (AP), allows parenchyma cells to sense a variety of blood-derived information, including osmotic ones. In the present study, we utilized immunohistochemistry to examine changes in the expression of NG2 and platelet-derived growth factor receptor beta (PDGFRB) in the OVLT, SFO and AP of adult mice during chronic osmotic stimulation. The expression of NG2 and PDGFRB was remarkably prominent in pericytes, although these angiogenesis-associated proteins are highly expressed at pericytes of developing immature vasculature. The chronic salt loading prominently increased the expression of NG2 in the OVLT and SFO and that of PDGFRB in the OVLT, SFO and AP. The vascular permeability of low-molecular-mass tracer fluorescein isothiocyanate was increased significantly by chronic salt loading in the OVLT and SFO but not AP. In conclusion, the present study demonstrates changes in pericyte expression of NG2 and PDGFRB and vascular permeability in the sensory CVOs by chronic osmotic stimulation, indicating active participation of the vascular system in osmotic homeostasis.

The viral mimetic polyinosinic:polycytidylic acid (poly I:C) induces cellular responses in primary cultures from rat brain sites with an incomplete blood-brain barrier.

Primary microcultures of the organum vasculosum laminae terminalis (OVLT) and the area postrema (AP), brain sites with an incomplete blood-brain barrier, were established from topographically excised rat pup tissue, with cellular identification by marker protein-specific immunocytochemistry. Employing the ratio calcium imaging technique, we showed for the first time that polyinosinic:polycytidylic acid (poly I:C) can induce calcium signalling in single OVLT and AP cells. Poly I:C stimulation caused fast, transient rises in intracellular calcium in about 5% of neurons and astrocytes and some microglial cells. Frequently, the responses of astrocytes and microglial cells showed a shorter onset-latency compared to neurons. In addition, exposure to poly I:C led to a time dependent release of bioactive tumour necrosis factor (TNF) and interleukin-6 (IL-6) into the supernatants of OVLT and AP cultures. The demonstration of direct cellular responses of OVLT- and AP-intrinsic cells to stimulations with poly I:C is in agreement with the discovered existence of Toll-like receptor 3 (TLR3), the cognate receptor for poly I:C, in the brain.

Sensory circumventricular organs in health and disease.

Circumventricular organs (CVOs) are specialized brain structures located around the third and fourth ventricles. They differ from the rest of the brain parenchyma in that they are highly vascularised areas that lack a blood-brain barrier. These neurohaemal organs are classified as "sensory", when they contain neurons that can receive chemical inputs from the bloodstream. This review focuses on the sensory CVOs to describe their unique structure, and their functional roles in the maintenance of body fluid homeostasis and cardiovascular regulation, and in the generation of central acute immune and febrile responses. In doing so, the main neural connections to visceral regulatory centres such as the hypothalamus, the medulla oblongata and the endocrine hypothalamic-pituitary axis, as well as some of the relevant chemical substances involved, are described. The CVOs are vulnerable to circulating pathogens and can be portals for their entry in the brain. This review highlights recent investigations that show that the CVOs and related structures are involved in pathological conditions such as sepsis, stress, trypanosomiasis, autoimmune encephalitis, systemic amyloidosis and prion infections, while detailed information on their role in other neurodegenerative diseases such as Alzheimer's disease or multiple sclerosis is lacking. It is concluded that studies of the CVOs and related structures may help in the early diagnosis and treatment of such disorders.

Aquaporin-1 in blood vessels of rat circumventricular organs.

Although the water channel protein aquaporin-1 (AQP1) is widely observed outside the rat brain in continuous, but not fenestrated, vascular endothelia, it has not previously been observed in any endothelia within the normal rat brain and only to a limited extent in the human brain. In this immunohistochemical study of rat brain, AQP1 has also been found in microvessel endothelia, probably of the fenestrated type, in all circumventricular organs (except the subcommissural organ and the vascular organ of the lamina terminalis): in the median eminence, pineal, subfornical organ, area postrema and choroid plexus. The majority of microvessels in the median eminence, pineal and choroid plexus, known to be exclusively fenestrated, are shown to be AQP1-immunoreactive. In the subfornical organ and area postrema in which many, but not all, microvessels are fenestrated, not all microvessels are AQP1-immunoreactive. In the AQP1-immunoreactive microvessels, the AQP1 probably facilitates water movement between blood and interstitium as one component of the normal fluxes that occur in these specialised sensory and secretory areas. AQP1-immunoreactive endothelia have also been seen in a small population of blood vessels in the cerebral parenchyma outside the circumventricular organs, similar to other observations in human brain. The proposed development of AQP1 modulators to treat various brain pathologies in which AQP1 plays a deleterious role will necessitate further work to determine the effect of such modulators on the normal function of the circumventricular organs.

Clinical and surgical features of lower brain stem hemangioblastomas in von Hippel-Lindau disease.

BACKGROUND: In the context of von Hippel-Lindau disease (VHL), the medulla oblongata is a relatively frequent site of growth of hemangioblastomas, posing related clinical and surgical difficulties. Their management requires a close correlation between clinical evolution and morphological surveillance. In order to describe their clinical and surgical features, we reviewed our experience in the treatment of these lesions. METHODS: Between 2001 and 2009, 14 patients (9 female and 5 male, mean age 34 years) underwent removal of 15 lower brain stem hemangioblastomas. Based on the review of the clinical records and outpatient long-term follow-up visits, their clinical course was analyzed. Functional evaluation was measured with the Karnofsky Performance Scale (KPS) on admission, at discharge and at the last follow-up. The mean follow-up period was 30.8 months (range 4-99). RESULTS: All the operated hemangioblastomas were located in the dorsal medulla oblongata, in the context of multiple lesions, cerebellar and/or spinal. In ten patients hemangioblastomas were located in a median position at the obex area; in four cases a lateral location was observed. Cystic component was absent in two cases. Clinical onset preceded surgery by a mean of 8.5 months. Preoperatively three patients showed a KPS lower than 80, ten patients between 80 and 90, and one patient scored 100 (asymptomatic). There was no surgical mortality. Nine out of 14 patients showed a temporary surgical morbidity. One patient required a tracheostomy. At follow-up ten patients scored a KPS better than before the operation, while the other four patients remained stable. Permanent morbidity was observed in three patients. CONCLUSIONS: Lower brain stem hemangioblastomas in the context of VHL show an often gradual onset of signs and symptoms except for patients who develop an obstructive hydrocephalus. Although transient surgical complications are possible, surgery provides favorable long-term results.

Development of the area postrema: an immunohistochemical study in the macaque.

The organization and chemical development of the area postrema (AP) in the macaque monkey was studied by immunohistochemistry imaged with conventional and confocal microscopy from day 40 of gestation to adulthood. The thin ependyma of the adult was found to develop from a thick continuous structure beginning in the second trimester. It was later invaded by tyrosine hydroxylase immunoreactive (TH+) and dopamine beta-hydroxylase immunoreactive (DBH+) cells and fibers, suggesting a possible route for release of neurotransmitter directly into ventricular cerebrospinal fluid. Other TH+ and/or DBH+ fibers were found in close approximation to blood vessels. Prominent vascularity of the parenchyma of AP was present late in the first trimester (fetal day (Fd)57 in the macaque) and increased further until birth. By contrast, the underlying solitary nucleus was hypervascular at Fd57, but its vascularity rapidly declined by late in the second trimester. TH+ neurons first appeared late in the first trimester, and DBH+ neurons appeared in the second trimester; these findings are consistent with the view that catecholaminergic cells in AP are the earliest members of the A2 noradrenergic group. Catecholaminergic cells or fibers in AP contained little labeling for synaptic vesicular proteins, suggesting that the release of neurotransmitter there may not involve a synaptic mechanism. Synapses were first observed in mid-second trimester, and most were associated with GABA+ fibers.

The circumventricular organs: an atlas of comparative anatomy and vascularization.

The circumventricular organs are small sized structures lining the cavity of the third ventricle (neurohypophysis, vascular organ of the lamina terminalis, subfornical organ, pineal gland and subcommissural organ) and of the fourth ventricle (area postrema). Their particular location in relation to the ventricular cavities is to be noted: the subfornical organ, the subcommissural organ and the area postrema are situated at the confluence between ventricles while the neurohypophysis, the vascular organ of the lamina terminalis and the pineal gland line ventricular recesses. The main object of this work is to study the specific characteristics of the vascular architecture of these organs: their capillaries have a wall devoid of blood-brain barrier, as opposed to central capillaries. This particular arrangement allows direct exchange between the blood and the nervous tissue of these organs. This work is based on a unique set of histological preparations from 12 species of mammals and 5 species of birds, and is taking the form of an atlas.

Microvascular patterns in human medullary tegmentum at the level of the area postrema.

Abstract The aim of the present study was to evaluate the regional differences in microvessel density (MVD) of the human medullary tegmentum in adults and newborns/infants. Transverse serial sections of formalin-fixed, paraffin-embedded brainstems, taken from 16 adult and eight newborn/infant subjects, were stained with anti-von Willebrand factor (vWF) polyclonal antibodies. The boundaries of the area postrema (AP), dorsal motor vagal nucleus (DMVN), solitary tract nucleus (STN), solitary tract (ST) and hypoglossal nucleus (XII) were defined, all vessels were counted and the values were divided by the areas. In adult cases, statistically significant heterogeneity in MVD was found among the nuclei studied (P < 0.001). DMVN and AP showed higher MVD with respect to XII and ST (P < 0.001). The MVD of STN was lower with respect to DMVN (P < 0.001) and higher with respect to XII and ST (P < 0.05). The MVD and capillary density of the AP of newborns/infants were not significantly different with respect to adults. In sections of the medulla of adult subjects stained with anti-vWF, all vessels showed an intense reaction of endothelial cells whereas in the DMVN, XII, STN and ST of newborns/infants, only rare, isolated vessels showed anti-vWF reactivity and in the AP, 41 +/- 21% of vessels expressed vWF. Differences in MVD among the nuclei may be related to their different functions and metabolic demands. Light and heterogeneous expression of vWF in endothelial cells of newborns/infants indicates that differentiation of microvasculature in the human medullary tegmentum extends beyond fetal stages.

Membrane-transport systems in the fenestrated capillaries of the area postrema in rat and calf.

The capillaries of the area postrema (AP) lack the morphological peculiarity of the blood-brain barrier (BBB), and the AP neurons are considered located outside the BBB. Using the immunofluorescent method, we have investigated the expression of membrane transport systems that are instrumental to the BBB function, such as caveolin-1, -2, P-glycoprotein, and glut-4, in the capillary endothelium of the rat and calf AP. The expression of these molecules was verified after fibronectin immunostaining of the microvessels. Both in the rat and calf, caveolin-1, -2, and P-glycoprotein were expressed in the AP capillaries. A quantitative analysis revealed that the proportion of the capillary profiles expressing these transport systems was very close to 100% of the fibronectin immunolabelled profiles. On the contrary, none of the AP capillaries showed glut-4 immunoreactivity. The present investigation demonstrates that the endothelial layer of the AP capillaries, in spite of the paracellular passage of polar molecules through the leaky tight junctions and fenestrations, could be an active interface which is able to control the entry of a wide range of blood-borne compounds into the brain by means of specific mechanisms, including an efflux pump.

The distribution of mast cells in the human area postrema.

The topography and phenotype of mast cells in the human area postrema, together with correlation between mast-cell density and microvessel density (MVD), were analysed in 16 brains. Transverse serial sections of formalin-fixed, paraffin-embedded brainstems were stained with toluidine blue and alcian blue/safranin stainings, and with anti-tryptase and anti-CD31 monoclonal antibodies. The mean (+/- SD) numbers of mast cells per section were 1.3 +/- 0.8 and 1.2 +/- 0.7 with toluidine blue and alcian blue/safranin, respectively, whereas anti-tryptase monoclonal antibody showed a mean of 5.1 +/- 2.4 cells. Mast cells were alcian blue- and safranin-positive in 56%, because of the coexistence of low-sulphated (blue-staining) and high-sulphated (red-staining) granules. No significant linear correlation between mast-cell density (4.9 mm(-2)) and MVD (114.5 mm(-2)) was found (r(2) = 0.19, P = 0.09). Mast cells were frequently located close to blood vessels (55%) (33% to venules, 22% to arterioles), indicating that their products play a role in the regulation of blood flow and in vessel permeability in the area postrema. Mast cells were located subependymally in 44% and close to the dorsal aspect of the nucleus of the tractus solitarius in 31%, suggesting a subregional distribution.