[Regulation of vasomotor tone of small skeletal muscle veins by intrinsic mechanisms]. / Vázizomkisvénák vazomotortónusának intrinszik szabályozómechanizmusai.

In many developed countries the prevalence of venous disorders and its consequences are higher than that of arterial diseases. Thus it is very important to understand the exact physiological and pathophysiological function of small veins and their control mechanisms. Small veins and venules have an important role in the regulation of capillary fluid exchange, as well as return of the venous blood into the heart. However, there is only limited knowledge available regarding the role of local mechanisms controlling the vasomotor tone and diameter of small veins. In the last decade the authors focused on the elucidation of these mechanisms in isolated skeletal muscle venules of rats. Their results suggest that the tone of small veins is controlled by the integration of several mechanisms, activated by the intraluminal pressure and flow/wall shear stress, in addition to numerous local mediators synthesized and released from the smooth muscle and endothelium. These mechanisms are involved - in a complex manner - in the control of postcapillary resistance, thus regulation of tissue blood supply, venous return and consequently in the modulation of the cardiac output, as well.

Neurohumoral regulation of spontaneous constrictions in suburothelial venules of the rat urinary bladder.

Venules of the bladder suburothelium develop spontaneous phasic constrictions that may play a critical role in maintaining venular drainage of tissue metabolites. We aimed to investigate neurohumoral regulation of the spontaneous venular constrictions (SVCs). Changes in venular diameter of the rat bladder suburothelium were monitored using a video tracking system, whilst the effects of electrical field stimulation (EFS) and bath-applied bioactive substances were investigated. The innervation of the suburothelial microvasculature was examined by immunohistochemistry. EFS (10Hz for 30s) induced an increase in the frequency of SVCs that was prevented by phentolamine (1µM). In phentolamine-pretreated venules, EFS suppressed SVCs with a venular dilatation in a manner attenuated by propranolol (1µM) or l-nitro arginine (LNA, 10µM). BRL37344 (1µM), a ß3 adrenoceptor agonist, dilated venules and reduced the frequency of SVCs in an LNA-sensitive manner. ACh (1-10µM) increased the frequency of SVCs. ATP (1µM) transiently constricted venules and then caused LNA-sensitive cessation of SVCs associated with a dilatation. Substance P (100nM) caused a venular constriction, whilst calcitonin gene related peptide (CGRP, 100nM) caused a dilatation of venules and suppression of SVCs that were not inhibited by LNA. Immunohistochemical staining demonstrated sympathetic as well as substance P- and CGRP-containing nerves running along the venules. Spontaneous constrictions of suburothelial venules are accelerated by sympathetic α-adrenergic stimulation, but suppressed upon ß-adrenergic stimulation. In addition, suburothelial venular constrictions appear to be modulated by several bioactive substances that could be released from urothelium or suburothelial sensory nerves.

Properties of submucosal venules in the rat distal colon.

BACKGROUND AND PURPOSE: Venules within the gut wall may have intrinsic mechanisms for maintaining the circulation even upon the intestinal wall distension. We aimed to explore spontaneous and nerve-mediated contractile activity of colonic venules. EXPERIMENTAL APPROACH: Changes in the diameter of submucosal venules of the rat distal colon were measured using video microscopy. The innervation of the microvasculature was investigated using fluorescence immunohistochemistry. KEY RESULTS: Submucosal venules exhibited spontaneous constrictions that were abolished by blockers of L-type Ca(2+) channels (1 µM nicardipine), Ca(2+)-ATPase (10 µM cyclopiazonic acid), IP3 receptor (100 µM 2-APB), Ca(2+)-activated Cl(-) channels (100 µM DIDS) or store-operated Ca(2+) entry channels (10 µM SKF96365). Transmural nerve stimulation (TNS at 10 Hz) induced a phasic venular constriction that was blocked by phentolamine (1 µM, α-adrenoceptor antagonist) or sympathetic nerve depletion using guanethidine (10 µM). Stimulation of primary afferent nerves with TNS (at 20 Hz) or capsaicin (100 nM) evoked a sustained venular dilatation that was attenuated by calcitonin gene-related peptide (CGRP) 8-37 (2 µM), a CGRP receptor antagonist. Immunohistochemistry revealed sympathetic and primary afferent nerves running along submucosal venules. CONCLUSIONS AND IMPLICATIONS: Submucosal venules of the rat distal colon exhibit spontaneous constrictions that appear to primarily rely on Ca(2+) release from sarcoplasmic reticulum and subsequent opening of Ca(2+)-activated Cl(-) channels that trigger Ca(2+) influx through L-type Ca(2+) channels. Venular contractility is modulated by sympathetic as well as CGRP-containing primary afferent nerves, suggesting that submucosal venules may play an active role in regulating the microcirculation of the digestive tract.

Excessive peptidergic sensory innervation of cutaneous arteriole-venule shunts (AVS) in the palmar glabrous skin of fibromyalgia patients: implications for widespread deep tissue pain and fatigue.

OBJECTIVE: To determine if peripheral neuropathology exists among the innervation of cutaneous arterioles and arteriole-venule shunts (AVS) in fibromyalgia (FM) patients. SETTING: Cutaneous arterioles and AVS receive a convergence of vasoconstrictive sympathetic innervation, and vasodilatory small-fiber sensory innervation. Given our previous findings of peripheral pathologies in chronic pain conditions, we hypothesized that this vascular location may be a potential site of pathology and/or serotonergic and norepinephrine reuptake inhibitors (SNRI) drug action. SUBJECTS: Twenty-four female FM patients and nine female healthy control subjects were enrolled for study, with 14 additional female control subjects included from previous studies. AVS were identified in hypothenar skin biopsies from 18/24 FM patient and 14/23 control subjects. METHODS: Multimolecular immunocytochemistry to assess different types of cutaneous innervation in 3 mm skin biopsies from glabrous hypothenar and trapezius regions. RESULTS: AVS had significantly increased innervation among FM patients. The excessive innervation consisted of a greater proportion of vasodilatory sensory fibers, compared with vasoconstrictive sympathetic fibers. In contrast, sensory and sympathetic innervation to arterioles remained normal. Importantly, the sensory fibers express α2C receptors, indicating that the sympathetic innervation exerts an inhibitory modulation of sensory activity. CONCLUSIONS: The excessive sensory innervation to the glabrous skin AVS is a likely source of severe pain and tenderness in the hands of FM patients. Importantly, glabrous AVS regulate blood flow to the skin in humans for thermoregulation and to other tissues such as skeletal muscle during periods of increased metabolic demand. Therefore, blood flow dysregulation as a result of excessive innervation to AVS would likely contribute to the widespread deep pain and fatigue of FM. SNRI compounds may provide partial therapeutic benefit by enhancing the impact of sympathetically mediated inhibitory modulation of the excess sensory innervation.

Spatiotemporal distribution of neurovascular alignment in remodeling adult rat mesentery microvascular networks.

An emerging area of microvascular research focuses on the links between neural and vascular patterning. However, the functional dependence between vascular and neural growth in adult tissues remains underinvestigated. The objective of this study was to determine the spatial and temporal coordination between vascular and neural networks over a time course of adult microvascular growth. Mesentery tissues from adult male Wistar rats were harvested prior to stimulation, and 2, 10 and 30 days after angiogenesis stimulated by mast cell degranulation. Tissues were immunolabeled for PECAM (endothelial cell marker) and class III ß-tubulin (peripheral nerve marker). Neurovascular alignment was quantified per vessel category: arterioles (>20 µm), pre-capillary arterioles (10-20 µm), post-capillary venules (10-20 µm), venules (>20 µm), capillaries (<10 µm) and capillary sprouts. Neurovascular alignment along pre-capillary arterioles, capillaries, post-capillary venules and venules was decreased compared to unstimulated levels on days 2 and 10. These decreases inversely correlated with increases in vessel density per vessel category. By day 30, alignment either returned to unstimulated levels or was increased compared to day 10. These results suggest that neurovascular alignment arises after microvascular network growth and is present along arterioles, venules and even capillaries.

Oscillating neuro-capillary coupling during cortical spreading depression as observed by tracking of FITC-labeled RBCs in single capillaries.

Coupling between capillary red blood cell (RBC) movements and neuronal dysfunction during cortical spreading depression (CSD) was examined in rats by employing a high-speed camera laser-scanning confocal fluorescence microscope system in conjunction with our Matlab domain software (KEIO-IS2). Following microinjection of K(+) onto the surface of the brain, changes in electroencephalogram (EEG), DC potential and tissue optical density were all compatible with the occurrence of a transient spreading neuronal depression. RBC flow in single capillaries was not stationary. Unpredictable redistribution of RBCs at branches of capillaries was commonly observed, even though no change in diameter was apparent at the reported site of the capillary sphincter and no change of arteriolar-venule pressure difference was detected. There appeared to be a slow morphological change of astroglial endfeet. When local neurons were stunned transiently by K(+) injection, the velocity and oscillation frequency of RBCs flowing in nearby capillaries started to decrease. The flow in such capillaries was rectified, losing oscillatory components. Sluggish floating movements of RBCs in pertinent capillaries were visualized, with occasional full stops. When CSD subsided, RBC movements recovered to the original state. We postulate that neuronal depolarization blocks oscillatory signaling to local capillaries via low-shear plasma viscosity increases in the capillary channels, and a complex interaction between the RBC surface and the buffy coat on the capillary wall surface increases the capillary flow resistance. Then, when CSD subsides and oscillatory neuronal function is recovered, the normal physiological conditions are restored.

[Localization of the neurokinin 1 receptor in hip joints of patients with painful osteoarthritis]. / Die Lokalisation des Neurokinin 1-Rezeptors im Hüftgelenk von Patienten mit schmerzhafter Osteoarthrose.

INTRODUCTION: Recent studies on osteoarthritis have focused on nociceptive substance P (SP) containing afferent nerve fibres. The effects of SP are known to be mainly mediated by the tachykinin receptor neurokinin 1 (NK1-R). AIM: The aim of the present study was to demonstrate the NK1-R in human joint tissues. METHODS: The hip joint capsule of three patients with painful hip osteoarthritis (Group 1), three patients with femoral neck fracture showing no cartilage destruction (Group 2, controls) and the soft tissue of the fossa acetabuli of Group 1 were resected during hip arthroplasty implantation. The tissue samples were cut into small blocks and immersion-fixed in Zamboni's fixative. The specimens were frozen, cut into 50 microm sections and immunostained using a standard immunohistochemical staining protocol. RESULTS: In Groups 1 and 2 the NK1-receptor was localised in the wall of venous vessels, on Schwann cells of nerve bundles and on nerve fibres. In the osteoarthritis group the staining pattern was similar but the number of NK1-bearing cell structures seemed to be enhanced. CONCLUSIONS: The present study provides the first evidence of NK1-R in the human hip joint. In patients with painful osteoarthritis the density of NK1-R-positive cell structures seemed to be increased. The localisation of the NK1 receptor on different cell types suggests multiple effects of SP in normal and osteoarthritic joints.

[Adrenergic vascular reactivity in elderly hypertensive patients with orthostatic disorders]. / Osobennosti adrenergicheskoi reaktivnosti sosudov u bol'nykh gipertonicheskoi bolezn'iu pozhilogo vozrasta s ortostaticheskimi narusheniiami.

Vascular adrenergic reactivity was studied in 100 healthy persons and 222 elderly patients with stage II essential hypertension at high risk of complications. With age, the rate of orthostatic hypotension rises. This hypotension in the elderly hypertensive patients is caused by reduced adrenergic reactivity of venous microvessels.

Three-dimensional wall structure and the innervation of dental pulp blood vessels.

The involvement of neural components in plasma extravasation and blood flow in the dental pulp has been established by pharmacological and physiological studies. We review here the segmental constitution of pulp vessels and the possible involvement of neural components in both the contractility and permeability of the pulp vessels from a morphological viewpoint. Six vascular segments can be identified based on the morphology of peri-endothelial cells, such as smooth muscle cells and pericytes. These are: muscular arterioles, terminal arterioles, precapillary arterioles, capillaries, postcapillary venules, and collecting or muscular venules. The perivascular nerve forms a mesh with numerous terminal varicosities, some of which attach directly to arteriolar smooth muscle cells. This mesh can be seen by scanning electron microscopy, and indicates the important role of neural components in regulating the pulpal circulation. After administering norepinephrine (0.2 mg/kg/dog), the surface texture of the smooth muscle cells of pulp arterioles reveals marked irregularities, which are correlated with arteriolar contraction. The pericytes in larger postcapillary venules (diameter 20 microm or larger) also show irregularities, whereas no changes are seen in the pericytes of either smaller postcapillary venules or capillaries. The intercellular spaces of pericytes in the postcapillary venules are wide enough for leukocytes to pass through, and the occasional extravasation of leukocytes through venule walls can be seen under electron microscopy. The microvessels of healthy human dental pulp react weakly to selectins, indicating that apparently healthy dental pulp may be weakly inflamed. In rat dental pulp, CGRP-immunoreactive nerves and nerve terminals containing many granular vesicles supply the postcapillary venules more densely than the arterioles, which suggests the involvement of postcapillary venules in neurogenic inflammation in the dental pulp.

Innervation of human trigeminal nerve blood vessels.

Film preparations and histological sections of human trigeminal nerve impregnated with silver nitrate and treated according to Falck-Hillarp revealed a rich innervation within the intraneural blood vessels. Highly diverse and complex neural interconnections were noted in the arterioles and venules of the trigeminal ganglion capsule as well as in the epineurium and external layers of the perineurium of the trigeminal nerve branches. Neural plexuses were detected on the vessel walls within these layers. Sensory innervation of the neural blood vessels was mainly relegated to polyvalent tissue-vascular receptors with adrenergic neural plexuses encountered on the walls of the intraneural vessels.

Heterogeneity of vascular innervation in hamster cheek pouch and retractor muscle.

The hamster cheek pouch and its retractor muscle have provided valuable insights into microvascular physiology of an epithelial tissue and striated muscle, respectively. Nevertheless, the innervation of these vascular beds has not been resolved. This study has investigated the nature of autonomic and sensory innervation of these vascular beds and has tested whether it varies within or between tissues. Multiple-labelling immunohistochemistry identified autonomic and peptide-containing sensory nerve fibres. Presumptive sympathetic vasoconstrictor axons with immunoreactivity (IR) for tyrosine hydroxylase (TH) and neuropeptide Y (NPY) innervated feed arteries and arterioles (but not veins or venules) of the retractor and anterior (muscular) cheek pouch; these axons were absent from the posterior (epithelial) region of the cheek pouch, as confirmed by catecholamine fluorescence. Presumptive autonomic vasodilator axons with IR for vasoactive intestinal peptide (VIP) consistently innervated feed arteries and proximal arterioles of the cheek pouch, but generally not those of the retractor muscle nor distal arterioles of either tissue. Sparse presumptive sensory axons with IR for calcitonin gene-related peptide (CGRP) and substance P were found near arterial and venous vessels in all regions of the cheek pouch and retractor muscle; CGRP-IR was also located in motor end plates associated with striated muscle fibres. Such regional differences in vascular innervation by autonomic and sensory neurons may selectively effect local and regional control of blood flow between and within vascular beds.

An immunohistochemical study on the innervation of two peptidergic (NPY and VIP) nerves in the cerebral arterial tree and choroid plexus of the newt (Amphibia: Urodela).

The pattern of neuropeptide Y (NPY)- and vasoactive intestinal polypeptide (VIP)-immunoreactive (IR) innervation was investigated in the cerebral arterial tree and choroid plexus of the newt by the use of an indirect immunofluorescence technique combined with chemical sympathectomy (6-OHDA). The data presented here, in conjunction with histochemical data reported previously, showed the following characteristic features of cerebrovascular innervation in this urodelan species. (1) The cerebral arterial tree and choroid plexus had unbalanced NPY-IR and VIP-IR innervation, characterized by the absence or a markedly lesser density of VIP-IR nerves. (2) All or nearly all of the NPY-IR nerves were sympathetic in nature. (3) A few cerebral perivascular NPY-IR nerves in some individuals originated from the sympathetic NPY-IR nerve cells intrinsic to the major cerebral arteries of the anterior circulatory system. (4) Acetylcholinesterase-positive neurons lacking both NPY and VIP immunoreactivities are a major nerve type in cerebrovascular parasympathetic innervation. The preferential NPY-IR innervation of the plexus microvascular-epithelial regions must be considered in relation to its special functions, such as the regulation of microcirculation, and cerebrospinal fluid (CSF) production and transportation. CSF is vital for the movement of nutrients and metabolites in the newt brain.

An immunohistochemical and ultrastructural study of vasomotor nerves in the microvasculature of human dental pulp.

Immunohistochemical methods for S-100 protein and neurone-specific enolase showed two types of nerve endings around the pulp microvasculature: the free endings, which comprise the major neural component and are distributed in all types of microvessel such as arterioles, venules and capillaries; and the varicose endings. The varicose ending was a less frequent, minor component observed only in the arterioles. Both immunohistochemical and ultrastructural observations confirmed that the varicose endings were the terminal axons of efferent vasomotor nerves. Further extensive ultrastructural examinations on the vasomotor nerves added the following new findings to our previous reports. Vasomotor nerves sometimes ramified into more than two terminal axons around arterioles, and most of these ramified axons ended in the adventitia-media junction of the arteriolar wall; however, nerve endings occasionally penetrated into the media. These findings suggest an intimate structural association between vasomotor nerves and arterioles in regulating the arteriolar microcirculation in human dental pulp by stimulating smooth-muscle cells not only of the outermost but also of the inner layers. Furthermore, the deep penetration of terminal axons into the arteriolar wall seems to provide effective regulation of pulpal blood flow under physiological and pathological conditions.

Ultrastructure of the neuromuscular junction of vasomotor nerves in the microvasculature of human dental pulp.

Vasomotor nerves in human dental pulp were more closely related to arterioles than to venules. Most were composed of unmyelinated fibres, which were mainly adrenergic. They appeared close to arterioles that were surrounded by a few layers of contractile smooth-muscle cells. The smaller arterioles with a diameter of 10-15 microns received a more intimate innervation by vasomotor nerves than did the larger. These vessels occasionally showed much narrower neuromuscular junctions than previously reported. Most of these nerve fibres were identified as adrenergic by the presence of chromaffin-positive synaptic vesicles detected by ultrastructural enzyme histochemistry. Their function appeared to be to regulate the blood flow and/or the blood pressure by stimulating smooth-muscle cells, resulting in contraction and a change in the calibre of the vessels. Capillaries and venules, which have a higher permeability, received weaker innervation by the vasomotor nerves than did arterioles. The intimate relation between vasomotor nerves and arterioles is related to the function of dental pulp in normal and pathological conditions.

[The pattern-dependent mechanisms of the neural regulation of the microvascular tonus in the rat stomach]. / Pattern-zavisimye mekhanizmy nervnoi reguliatsii tonusa mikrososudov zheludka krys.

The effects of electrical stimulation of the left splanchnic nerve in 1 s bursts at 4 s intervals at 5-80 Hz or continuously at 1-16 Hz (6 V, 1 ms) on gastric submucosal microvasculature were studied by the reflected light in vivo TV-microscopy. Burst stimulation, like a continuous one, induced frequency-dependent vasoconstrictor responses. Maximal reduction in diameter of both arterioles (by about 90%) and venules (by about 60%) occurred at the same total number of stimuli delivered for 1-min period of nerve stimulation continuously at 16 Hz or in bursts at 80 Hz. In other frequencies used, burst stimulation evoked significantly more pronounced contractile responses of arterioles (but not the venules), which followed by much less pronounced autoregulatory escape and poststimulatory hyperemia as compared to continuous stimulation at the comparable total number of impulses. After alpha-adrenoreceptor blockade, reduced in magnitude and slow developing contractile responses of arterioles persisted to stimulation in bursts rather than continuously suggesting the involvement of nonadrenergic co-transmitters) release. The data obtained show that not only the number of neural pulses but also their bursting pattern may have an informational role in microvascular contractile responses. Bursting pattern of nerve stimulation seems to be more "physiological" than the continuous one.

Sinusoidal vessels in the periodontal ligament of hamster incisors: their distribution, structure and possible function.

Vascular architecture in the periodontal ligament of hamster incisors was investigated by use of vascular casts under a scanning electron microscope (SEM). In addition to ordinary nutrient blood vessels, anastomosing vessels of large caliber developed, surrounding the incisor. From their characteristic configuration, these vessels were regarded as "sinusoids". The plexus of sinusoidal vessels was connected with capillaries in the papillary layer of the enamel organ at the labial periodontal ligament, and with veins penetrating into the alveolar bone on the lingual side. Transmission electron microscopic (TEM) observation showed that the sinusoidal wall was composed of only a thin layer of endothelial cells, lacking a smooth muscular element, and surrounded by densely arranged collagen fibers. Although the frequent association of Ruffini-type nerve endings with sinusoidal vessels was noted, neither direct contact nor specialized structures between these was recognizable. A possible function of the periodontal sinusoids is discussed on the basis of their distribution and ultrastructural evidence.

Substance P-immunoreactive sensory axons in the rat respiratory tract: a quantitative study of their distribution and role in neurogenic inflammation.

Substance P is one of the peptides released from sensory nerves that mediate "neurogenic inflammation." Although substance P-immunoreactive (SP-IR) axons are known to be present within the mucosa of the respiratory tract, the relative extent of the innervation of various components of the mucosa is not known. Therefore, we determined the distribution and number of SP-IR axons in the rat trachea and bronchi, by using immunohistochemistry on tissue whole mounts. Specifically, we sought to learn whether these axons directly innervate the postcapillary venules involved in neurogenic plasma extravasation, the arterioles involved in neurogenic vasodilatation, and the airway smooth muscle involved in bronchoconstriction in pathogen-free, adult male F344 rats. We found that 90% of the SP-IR axons were single axons, usually having varicosities. Eighty-five percent of these were in the epithelium, 6% innervated arterioles, and the remainder elsewhere in the lamina propria. Only 10% of the mediator-sensitive postcapillary venules (i.e., venules labeled with Monastral blue pigment after challenge with capsaicin or substance P) were within 10 microns of SP-IR axons. SP-IR axons were more than 10 times as frequent in the smooth muscle of the distal bronchi as in the trachea. Capsaicin pretreatment (168 mg/kg over 7 days) reduced the number of SP-IR axons in the trachea by 96%, which is consistent with their being sensory. Unilateral vagotomy reduced the number of SP-IR axons bilaterally in the trachea and ipsilaterally in the main stem bronchus. Using an antibody to Protein Gene Product 9.5 as a nonspecific marker for all nerves in the trachea, we determined that SP-IR axons constituted 90% of the axons in the epithelium, 32% of the axons on arterioles, and only 4% of the axons in the smooth muscle. We conclude that most SP-IR nerves in the trachea are sensory axons and most of these axons end in the epithelium. SP-IR axons innervate mucosal arterioles, but few innervate postcapillary venules. Therefore, the mechanism by which sensory axons evoke plasma extravasation from these venules is likely to involve the diffusion of the peptide or a secondary mediator from the epithelium or from the arterioles upstream.