Inverse neurovascular coupling contributes to positive feedback excitation of vasopressin neurons during a systemic homeostatic challenge.

Neurovascular coupling (NVC), the process that links neuronal activity to cerebral blood flow changes, has been mainly studied in superficial brain areas, namely the neocortex. Whether the conventional, rapid, and spatially restricted NVC response can be generalized to deeper and functionally diverse brain regions remains unknown. Implementing an approach for in vivo two-photon imaging from the ventral surface of the brain, we show that a systemic homeostatic challenge, acute salt loading, progressively increases hypothalamic vasopressin (VP) neuronal firing and evokes a vasoconstriction that reduces local blood flow. Vasoconstrictions are blocked by topical application of a VP receptor antagonist or tetrodotoxin, supporting mediation by activity-dependent, dendritically released VP. Salt-induced inverse NVC results in a local hypoxic microenvironment, which evokes positive feedback excitation of VP neurons. Our results reveal a physiological mechanism by which inverse NVC responses regulate systemic homeostasis, further supporting the notion of brain heterogeneity in NVC responses.

Neuronal-derived nitric oxide and somatodendritically released vasopressin regulate neurovascular coupling in the rat hypothalamic supraoptic nucleus.

The classical model of neurovascular coupling (NVC) implies that activity-dependent axonal glutamate release at synapses evokes the production and release of vasoactive signals from both neurons and astrocytes, which dilate arterioles, increasing in turn cerebral blood flow (CBF) to areas with increased metabolic needs. However, whether this model is applicable to brain areas that also use less conventional neurotransmitters, such as neuropeptides, is currently unknown. To this end, we studied NVC in the rat hypothalamic magnocellular neurosecretory system (MNS) of the supraoptic nucleus (SON), in which dendritic release of neuropeptides, including vasopressin (VP), constitutes a key signaling modality influencing neuronal and network activity. Using a multidisciplinary approach, we investigated vasopressin-mediated vascular responses in SON arterioles of hypothalamic brain slices of Wistar or VP-eGFP Wistar rats. Bath-applied VP significantly constricted SON arterioles (Δ-41 ± 7%) via activation of the V1a receptor subtype. Vasoconstrictions were also observed in response to single VP neuronal stimulation (Δ-18 ± 2%), an effect prevented by V1a receptor blockade (V2255), supporting local dendritic VP release as the key signal mediating activity-dependent vasoconstrictions. Conversely, osmotically driven magnocellular neurosecretory neuronal population activity leads to a predominant nitric oxide-mediated vasodilation (Δ19 ± 2%). Activity-dependent vasodilations were followed by a VP-mediated vasoconstriction, which acted to limit the magnitude of the vasodilation and served to reset vascular tone following activity-dependent vasodilation. Together, our results unveiled a unique and complex form of NVC in the MNS, supporting a competitive balance between nitric oxide and activity-dependent dendritic released VP, in the generation of proper NVC responses.

Hypothalamic cocaine- and amphetamine-regulated transcript and corticotrophin releasing factor neurons are stimulated by extracellular volume and osmotic changes.

Several studies suggest that hypothalamic cocaine- and amphetamine-regulated transcript (CART) may interact with the hypothalamic-pituitary-adrenal (HPA) axis in the control of neuroendocrine function and may also participate in cardiovascular regulation. Therefore, this study aimed to evaluate, in experimental models of isotonic (I-EVE) and hypertonic (H-EVE) extracellular volume expansion and water deprivation (WD), the activation of CART- and corticotrophin releasing factor (CRF)-immunoreactive neurons, as well as the relative expression of CART and CRF mRNAs in the paraventricular (PVN) and supraoptic (SON) nuclei of the hypothalamus. Both H-EVE (0.30M NaCl, 2mL/100g of body weight, in 1 minute) and 24 hours of WD significantly increased plasma sodium concentrations, producing, respectively, either an increase or a decrease in extracellular volume. I-EVE (0.15M NaCl, 2mL/100g of body weight, in 1 minute) evoked a significant increase in the circulating volume accompanied by unaltered plasma concentrations of sodium. CART-expressing neurons of both magnocellular and parvocellular hypothalamic divisions were activated to produce Fos in response to H-EVE but not in response to I-EVE. Furthermore, increased expression of CART mRNA was found in the PVN of H-EVE but not I-EVE rats. These data show for the first time that EVE not only activates hypothalamic CRF neurons but also increases CRF mRNA expression in the PVN. In contrast, WD increases the number of CART-immunoreactive neurons activated to produce Fos in the PVN and SON but does not change the number of neurons double labeled for Fos and CRF or expression of CRF mRNA in the PVN. These findings provided new insights into the participation of CART in diverse processes within the PVN and SON, including its possible involvement in activation of the HPA axis and cardiovascular regulation in response to changes in extracellular volume and osmolality.

Hypervascularization in the magnocellular nuclei of the rat hypothalamus: relationship with the distribution of aquaporin-4 and markers of energy metabolism.

In the magnocellular nuclei of the hypothalamus, there is a rich vascular network for which the function remains to be established. In the supraoptic nucleus, the high vascular density may be one element, which together with the water channel aquaporin-4 expressed in the astrocytes, is related to a role in osmoreception. We tested the osmoreception hypothesis by studying the correlation between vascular and cellular densities in the paraventricular nucleus and the supraoptic nucleus. Whether aquaporin-4 is likely to contribute to osmoreception was tested by studying the distribution in the magnocellular nuclei of the hypothalamus. The high vascular density may also reflect a high metabolic activity due to the synthesis of vasopressin and oxytocin. This metabolic hypothesis was tested by studying the regional cytochrome oxidase histochemistry, the local cerebral blood flow, and the density of glucose transporter type-1 in the supraoptic and paraventricular nuclei. All the magnocellular nuclei were characterized by an extended and intense aquaporin-4 labelling and a weak cytochrome oxidase histochemistry. The highest vascular density was found in the supraoptic nucleus and the magnocellular regions of the paraventricular nucleus. The local cerebral blood flow rates were surprisingly low in the paraventricular nucleus and the supraoptic nucleus in comparison to the cerebral cortex. Furthermore in these nuclei, the antibody for glucose transporter type-1 revealed two populations of vessels differing by their labelling intensity. The similarities observed between the different nuclei suggest that, in the hypothalamus, all magnocellular regions sense the plasma osmolarity. The low local cerebral blood flow, and the patterns of glucose transporter type-1 labelling and cytochrome oxidase histochemistry suggest that the high vascularization of these hypothalamic nuclei is not related to a high metabolic capacity in basal conditions.

Galanin immunoreactivity in mouse basal forebrain: sex differences and discrete projections of galanin-containing cells beyond the blood-brain barrier.

The distribution of galanin-immunoreactive (GAL-IR) cell bodies in the basal forebrain of mice was investigated. The overall pattern of staining for GAL in the area of brain analyzed was similar to that reported in other species with noticeable variations. Distinctive groups of GAL-IR cells were present in the bed nucleus of stria terminalis (BNST), supraoptic nucleus, retrochiasmatic supraoptic nucleus (SOR), magnocellular paraventricular nucleus, arcuate nucleus (ARC) and the nucleus circularis which is one of the cell groups belonging to the accessory magnocellular system. Comparison of the number of GAL-IR cells between the sexes indicated sexual dimorphism in the BNST, SOR and the ARC. As compared with female mice, the mean number of GAL-IR cells/section in the BNST and the SOR was higher and that in the ARC was lower in the males. Unlike in rats, the preoptic area contained mostly scattered GAL-IR cell bodies. Intraperitoneal injection of the retrograde tracer fluoro-gold in male mice resulted in uptake of fluoro-gold by selective GAL-IR cell groups in the basal forebrain suggesting that only some of these cell groups may project outside the blood-brain barrier whereas others may be involved in intracerebral neural transmission.

Hypothalamic accessory nuclei and their relation to the angiotensinergic and vasopressinergic systems.

The existence and colocalization of angiotensin II- and vasopressin-like immunoreactivity in individual magnocellular cell groups of the hypothalamus has been demonstrated by using immunocytochemical methods. These neurosecretory magnocellular groups consist of the paraventricular nucleus and the supraoptic nucleus, as well as different accessory cell groups. The fibers from the neurons of the accessory nuclei project directly to adjacent blood vessels and do not comigrate with the hypothalamo-neurohypophysial fiber pathway. On the basis of these findings it can be concluded that in the hypothalamus two different angiotensinergic and vasopressinergic neurosecretory systems exist: (1) an intrinsic hypothalamic and (2) a hypothalamo-neurohypophysial system. The distribution of the accessory cell groups in the hypothalamus is shown in a 3D reconstruction which includes the connection of these magnocellular nuclei with the vascular system in this area.

Microvascular density of the human paraventricular nucleus decreases with aging but not hypertension.

Microvascular density was determined in the supraoptic nuclei (SON) and paraventricular nuclei (PVN) of fixed brain tissue from 19 human subjects ranging in age from 30 to 85 years. Eight of these patients had hypertension. No sex- or hypertension-related differences in microvascular density were found in either the SON or PVN. However, capillary density decreased in the PVN with aging. These results indicate a differential pattern of microvascular loss in the human hypothalamus with aging.

Vasopressinergic neurons and the associated blood vessels in the rat anterior hypothalamus: an immunohistochemical study.

The paraventricular (PVN) and supraoptic (SON) nuclei of the hypothalamic neurosecretory system have been extensively investigated by many workers. The functional aspects of vasopressin secretion (elaborated by the PVN and SON neurons) in relation to the vasculature of the anterior hypothalamus are also well documented. However, the available data concerning vasopressin (VP) functions are largely based on physiological studies. Corroborative morphological correlation with regard to this has received little attention. The present report elucidates the intricate anatomical relationships between the VP-neurons and the adjoining capillaries in the rat anterior hypothalamus. A peroxidase-antiperoxidase (PAP) immunocytochemical study, using a commercial VP antibody, was carried out for this purpose. The observations are interpreted from a functional standpoint. VP-immunostained elements, i.e. the somata and the processes (mainly dendrites), were localized (i) close to the wall, (ii) on the endothelium, and (iii) occasionally, in the lumen of the hypothalamic capillaries. The findings provide immunocytochemical evidence that the vasopressinergic elements are in direct relationship with the hypothalamic vasculature. This raises some interesting possibilities for the former to be involved in: (i) affecting the permeability of the blood-brain barrier for transport of various nutrient substances (important in aging and Alzheimer's disease), (ii) inducing an alteration in the water permeability of the brain vessels on which depends the precise adjustment of brain water content and of brain volume (fundamental to normal functioning of the brain), and (iii) serving as osmoreceptors of the blood flowing through the capillaries and thus providing a feedback mechanism for VP modulation.

[Electron-microscopic changes in neurosecretory nuclei of the rat hypothalamus after total ischemia in short-term clinical death]. / Zmiany mikroskopowo-elektronowe w jadrach neurosekrecyjnych podwzgórza szczura po calkowitym niedokrwieniu w sytuacji krótkotrwalej smierci klinicznej.

Hypothalamic neurosecretory nuclei of rats in which short-term clinical death was experimentally induced, were examined electron-microscopically. Material for studies was taken two weeks after resuscitation of animals following 5 min cardiac and respiratory arrest. Application of aldehyde fixatives with Alcian blue as well as fixatives containing tannic acid revealed numerous additional ultrastructural features not visualized in routine electron microscopy. The obtained results are indicative that short-term cerebral ischemia in rats results in ultrastructural abnormalities in neurosecretory hypothalamic nuclei, which appear after 2 weeks survival. These are electron microscope exponents of reduced secretory function, increased number of lysosomal structures and changes in cytoskeleton organization. They are accompanied by abnormalities in axoplasm and myelin sheaths. The authors consider those structural abnormalities as exponents of delayed irreversible metabolic changes of nerve cells.

Morphometry of individual capillary beds in the hypothalamo-neurohypophysial system of rats.

Unusually high capillary densities, volumes, and surface areas were revealed by morphometric analysis of the hypothalamic paraventricular nuclei (magnocellular and parvocellular divisions), supraoptic nuclei, and pituitary neural lobe of Long-Evans and Brattleboro rats. Capillaries of the magnocellular division of the paraventricular nucleus were greater in number, but smaller in diameter, than those in the parvocellular division. Expressed per unit area or volume, capillary dimensions in the hypothalamo-neurohypophysial system of Brattleboro rats were the same as in Long-Evans rats.

Differences in function and structure of the capillary endothelium in the supraoptic nucleus and pituitary neural lobe of rats. Evidence for the supraoptic nucleus as an osmometer.

The physiology and structure of capillary endothelial cells in the hypothalamic ventromedial and supraoptic nuclei and pituitary neural lobe were evaluated with quantitative methods and compared. The capillary endothelial cells in the ventromedial nucleus were used as an index of blood-brain barrier endothelium in cerebral gray matter; this endothelium has relatively low surface area and low permeability to tracer solutes. The permeability X surface area product of endothelial cells for a neutral amino acid, 14C-alpha-aminoisobutyric acid (AIB), in the ventromedial nucleus was similar to the value for supraoptic nucleus and was several hundred times smaller than in the neural lobe. The supraoptic nuclei and neural lobe had exceptionally large capillary surface areas, but dissimilar rates of blood flow and transendothelial influx of AIB. Differences in permeability of the endothelial cells between these two structures correlated closely with their marked dissimilarities in morphology. The neural lobe endothelium had numerous fenestrations (five per capillary cross-section) and vesicular profiles (twice as many as supraoptic nucleus), two features commonly associated with high capillary permeability. The capillary endothelium of the supraoptic nucleus was that of a typical blood-brain barrier structure having intercellular junctions that appeared tight, no fenestrations, and few cytoplasmic pits and vesicles. The unusually large capillary surface area of the supraoptic nucleus and low rate of solute flux across its endothelial cells make this nucleus a unique structure in which rapid changes in tissue volume may occur in response to small perturbations in plasma osmolality. The findings implicate the supraoptic nucleus as an osmotically sensitive detector or 'osmometer' in neuroendocrine regulation of body fluid homeostasis.

[Features of microcirculation in the neurosecretory nuclei of the human hypothalamus in sudden cardiac death]. / Osobennosti mikrotsirkuliatsii v neirosekretornykh iadrakh gipotalamusa cheloveka pri vnezapnoi serdechnoi smerti.

The microcirculatory bed of the neurosecretory nuclei was studied post-mortally in three groups of patients with various levels of the functional activity of the hypothalamus-pituitary neurosecretory system (HPNS), namely basal, high and lowered. Sudden death from a heart attack was associated with signs of inhibition in the HPNS cells, edema manifestations in the HPNS zone, by a considerable increase in the number of functionally inactive capillaries, narrowing of their lumina and a decrease in their transport capacities determined by the level of alkaline phosphatase activity in their wall. These changes correlated with an elevation of the number of dark cells in the neurosecretory nuclei which suggests the existence of the functional connection between them.

A combined vascular-catecholamine fluorescence method reveals the relative vascularity of rat locus coeruleus and the paraventricular and supraoptic nuclei of the hypothalamus.

Vascular perfusion with a fluorescent dye, Pontamine sky blue, was combined with glyoxylic acid-induced catecholamine fluorescence to produce simultaneous staining of intracerebral blood vessels and catecholamine-containing cells and fibers. Quantitative measurements of blood vessels per unit area revealed that the vascular density of rat locus coeruleus did not differ from the surrounding neuropil but was significantly lower than that of the hypothalamic supraoptic and paraventricular (PVN) nuclei. Moreover, the density of blood vessels in the magnocellular portion of PVN was not uniform throughout its rostrocaudal extent but was dense only in a specific subdivision of lateral PVN, which is predominantly vasopressinergic and is also heavily innervated by catecholamine terminals.

[Development of neuron-capillary interrelationships in the nuclei of the anterior portion of the human hypothalamus during ontogenesis]. / Osobennosti razvitiia neiron-kapilliarnykh vzaimootnoshenii v iadrakh perednego otdela gipotalamusa cheloveka v ontogeneze.

Three periods in the formation of the spatial arrangement of the neuronal nuclei have been stated: 1) from the prenatal period, infancy including--pronounced decrease in the density of the neuronal cells arrangement; 2) from the childhood up to adolescence--relative stabilization in the hypothalamic cellular composition; 3) mature and old age--secondary decrease in the dencity of the neuronal cells arrangement. Certain peculiarities in the nuclear capillary composition and in their neuronal-capillary relationships have been revealed: during the prenatal ontogenesis, the dencity of the neuronal cells arrangement decreases and their size increases that is followed by decrease in their capillary loops. In postnatal ontogenesis no proportional relationship between the dencity of neuronal cells arrangement and that of capillary network in the nuclei of the anterior hypothalamus was noted.

[Morphometric studies of the density and diameter of capillaries in supraoptic and paraventricular nuclei of unmilked and milked cows]. / Morphometrische Untersuchungen über die Kapillardichte und den Kapillardurchmesser im Nucleus supraopticus und Nucleus paraventricularis bei ungemolkenen und gemolkenen Kühen

With mathematical-statistical methods the density and the diameter of the capillaries of the NSO and NPV were investigated by unmilked and milked cows. After the milking the diameter of the capillaries is significantly expanded in both nuclei districts. The density of the capillaries measured through the length of the capillaries pro mm3 tissue, were not significantly enlarged after the milking stimulus. The adaptation on the changed functionstate of the neurosecreoty cells arrive at a maximum 15 min after the milking, to go back afterwards, by 60 min after the milking studied cows, to the values of the beginning. It is discussed, that the intensified blood maintenance of the activated neurosecretory cells of the NSO and NPV is of a higher oxygen consumption.

Blood supply of the rat hypothalamus. III. Anterior region of the hypothalamus (nucleus suprachiasmatis, nucleus hypothalamicus anterior, nucleus periventricularis).

The arterial and venous blood supply of the anterior hypothalamus was studied by means of double perfusion technique. The angiotopography and cytoarchitecture of the hypothalamus were compared on serial sections by accounting for the three-dimensional coordinates. A detailed description is given of the arteries and veins contributing to the blood supply of the suprachiasmatic, the anterior hypothalamic and the periventricular nuclei. The topography of the arterial and venous trunk on the base surface of the rat diencephalon is described, as well as that of the larger branches which enter from below and pass through the anterior hypothalamus.