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Velocity differences between L- and P-selectin-dependent neutrophil rolling in venules of mouse cremaster muscle in vivo.

After surgical trauma, leukocyte rolling is initially normal in L-selectin-deficient mice and reduced at later times, whereas leukocyte rolling is initially absent in P-selectin-deficient mice but induced later. Here, we examined the possibility that P- and L-selectin support rolling at different characteristic velocities using intravital microscopy of venules of the exteriorized cremaster muscle venules of wild type (WT) and P- and L-selectin-deficient mice. At > 50 min after exteriorization, rolling in P-selectin-deficient mice occurred at significantly higher velocities (129 +/- 89 microns/s) than in WT mice (49 +/- 23 microns/s). Rolling velocity distribution in L-selectin-deficient mice was similar to WT mice immediately after exteriorization. Histological examination of Giemsa-stained whole-mount preparations in cremaster muscle venules revealed that the majority of rolling cells (approximately 90% in all genotypes) were granulocytes. We conclude that P-selectin mediates leukocyte rolling at velocities < 50 microns/s, whereas L-selectin sustains more rapid rolling. Under physiological conditions, P- and L-selectin synergize to support rolling at velocities between 20 and 70 microns/s as seen in WT mice.

Abdominal Muscles↗

Compression of intramyocardial arterioles during cardiac contraction is attenuated by accompanying venules.

It was calculated how cardiac contraction influences the luminal cross-sectional area of a maximally dilated coronary arteriole (37-micron inner diameter at a pressure of 35 mmHg) that is accompanied by two equal venules (45-micron inner diameter at a pressure of 17 mmHg), forming a so-called "triad." It was found that, during a contraction with 14% cardiac muscle shortening, arteriolar area is virtually unaffected (increase of 4%) at the expense of a large (55%) decrease in venular area. For comparison, the areas of an unaccompanied arteriole and an unaccompanied venule were calculated to be reduced by 45 and 36%, respectively, demonstrating the "protective effect" on accompanied arterioles in a triad. During contraction, the overall resistance of a system consisting of one arteriole in series with two parallel venules of equal length was calculated to increase about twice as much for nonaccompanied vessels (resistance increases by a factor of 2.8) than for vessels in a triad arrangement (resistance increased by a factor of 1.4). The calculations show that the extravascular (intramyocardial) pressure, which determines vascular area, is not an independent variable as in the intramyocardial pump and waterfall models but depends on the vascular "loading" conditions. Thus the small venular pressure together with the large venular compliance causes the extravascular pressure to remain low during contraction, thereby protecting the stiff arteriole at high pressure. We conclude that the triad arrangement of intramyocardial coronary vessels attenuates the increase in coronary resistance during cardiac contraction and thus has an important functional advantage.

Arterioles↗

Diameter changes in skeletal muscle venules during arterial pressure reduction.

Previous studies in skeletal muscle have shown a substantial (>100%) increase in venous vascular resistance with arterial pressure reduction to 40 mmHg, but a microcirculatory study showed no significant venular diameter changes in the horizontal direction during this procedure. To examine the possibility of venular collapse in the vertical direction, a microscope was placed horizontally to view a vertically mounted rat spinotrapezius muscle preparation. We monitored the diameters of venules (mean diameter 73. 8 +/- 37.0 microm, range 13-185 microm) oriented horizontally and vertically with a video system during acute arterial pressure reduction by hemorrhage. Our analysis showed small but significant (P < 0.0001) diameter reductions of 1.0 +/- 2.5 microm and 1.8 +/- 3. 1 microm in horizontally and vertically oriented venules, respectively, upon reduction of arterial pressure from 115.0 +/- 26. 3 to 39.8 +/- 12.3 mmHg. The venular responses were not different after red blood cell aggregation was induced by Dextran 500 infusion. We conclude that diameter changes in venules over this range of arterial pressure reduction are isotropic and would likely increase venous resistance by <10%.

Animals↗

Erythrocyte margination and sedimentation in skeletal muscle venules.

Previous studies in skeletal muscle of the dog and cat have shown that venous vascular resistance changes inversely with blood flow and may be due mainly to red blood cell aggregation, a phenomenon present in these species. To determine whether red blood cell axial migration and sedimentation contribute to this effect, we viewed either vertically or horizontally oriented venules of the rat spinotrapezius muscle with a horizontally oriented microscope during acute arterial pressure reduction. With normal (nonaggregating) rat blood, reduction of arterial pressure did not significantly change the relative diameter of the red blood cell column with respect to the venular wall. After induction of red blood cell aggregation in the rat by infusion of Dextran 500, red blood cell column diameter decreased up to 35% at low pseudoshear rates (below approximately 5 s(-1)); the magnitude was independent of venular orientation. In vertically oriented venules, the plasma layer was symmetrical, whereas in horizontally oriented venules, the plasma layer formed near the upper wall. We conclude that, although red blood cell axial migration and sedimentation develop in vivo, they occur only for larger flow reductions than are needed to elicit changes in venous resistance.

Animals↗

Hydraulic conductivity of ischemic pulmonary venules.

We report the first determination of lung endothelial barrier properties in ischemic, nonreperfused microvessels. We quantified the endothelial barrier in terms of hydraulic conductivity (Lp) in single pulmonary venules (diameter 20-50 microns) of isolated blood perfused lungs (dog, rat), held at constant inflation pressure (5 cmH2O) with a gas mixture containing 21% oxygen. Lp were determined by our split-drop technique in which an oil drop is first microinjected into a venule and then split by microinjection of a protein solution. Lp was interpreted from measurements of the rate of oil drop movement. Baseline Lp recorded in the first 30 min of perfusion averaged 3.4 +/- 0.9 x 10(-7) ml/(cm2.s.cmH2O). Then, in two separate groups of venules in which we established 1.3 +/- 0.1 h and 3.4 +/- 0.8 h of ischemia, we determined Lp which were, respectively, 145 +/- 6.5 and 308 +/- 13% above baseline (P < 0.05). We conclude that ischemia alone, in the absence of reperfusion, significantly deteriorates the lung endothelial barrier.

Animals↗

Location of focal silver staining at endothelial gaps in inflamed venules examined by scanning electron microscopy.

The century-old histological technique of silver nitrate staining has proven to be extremely useful for visualizing endothelial cell borders and localizing endothelial gaps, but the significance of the staining is still not fully understood. To gain some insight into what silver nitrate stains, we developed a method that enabled us to use scanning electron microscopy with backscattered and secondary electron imaging to examine silver staining at endothelial cell borders of venules of the rat tracheal mucosa. We found that in normal venules, silver lines followed the smooth contour of cell borders. However, 1 min after endothelial permeability was increased by substance P, cell borders were irregular and displaced from the silver lines by as much as 4.3 microns, and the lines were accompanied by three types of silver deposits. Most common (46% of total) were annulus-shaped silver deposits that surrounded endothelial gaps. These deposits averaged 1.5 microns in width, were positioned symmetrically across cell borders, and were located at a depth of 0.3 micron beneath the luminal surface. Many endothelial gaps were partitioned into multiple pores (mean, 2.4) by fingerlike processes of endothelial cells. Surprisingly, the gaps occupied only 5.4% of the total area of the silver deposits and constituted 0.15% of the luminal surface of the leaky postcapillary venules. A second type of silver deposit (19% of total) was positioned asymmetrically with respect to the cell border and marked sites where endothelial cell margins still overlapped but appeared to be vertically separated by obliquely oriented gaps. A third type marked gaps at three-cell junctions; these were no more abundant than deposits elsewhere around the cell perimeter, suggesting that three-cell junctions were not unusually leaky sites. We conclude that silver nitrate marks endothelial cell borders and outlines endothelial cell gaps by staining an element of intercellular junctions. The annular shape of many silver deposits around gaps suggests that junctional elements in the apposing cells are separated during gap formation but are still present at the gap perimeter.

Animals↗

Hydraulic conductivity of lung venules determined by split-drop technique.

The split-drop method has been used to determine filtration rate per unit surface area in the single pulmonary venule. In isolated perfused lungs of nine dogs, blood flow was stopped at different vascular pressures. By means of a double-micropuncture technique under stereomicroscopy, an oil drop was injected in a subpleural venule. The oil drop was then split with a solution of albumin (5.6 g/100 ml) in Ringer lactate. As the Ringer-albumin solution filtered, the distance between the menisci of the split oil drop (split-drop length) decreased. The split-drop geometry and the rate of change of split-drop length were recorded. The calculated venular filtration rate per unit surface area related linearly with vascular pressure (P less than 0.05). The slope of the line equaled venular hydraulic conductivity, which averaged 2.9 +/- 0.02 x 10(-7) ml/(cm2.s.cmH2O). Hydraulic conductivity is lower in lung than in systemic venules.

Animals↗

Hypoxia causes leukocyte adherence to mesenteric venules in nonacclimatized, but not in acclimatized, rats.

Although the effects of ischemia-reperfusion have received considerable attention, few studies have directly evaluated the microcirculatory response to systemic hypoxia. The overall objective of this study was to assess the effect of environmental hypoxia on adhesive interactions of circulating leukocytes with rat mesenteric venules by using intravital microscopy. Experiments were designed to 1) characterize the adhesive interactions of circulating leukocytes to venules during acute hypoxia produced by a reduction in inspired PO(2), 2) evaluate the role of nitric oxide in these adhesive interactions, 3) determine whether the effect of hypoxia on leukocyte adhesive interactions differs between acclimatized and nonacclimatized rats, and 4) assess whether compensatory changes in nitric oxide formation contribute to this difference. The results showed that acute hypoxia promotes leukocyte-endothelial adherence in mesenteric venules of nonacclimatized rats. The mechanism of this response is consistent with depletion of nitric oxide within the microcirculation. In contrast, no leukocyte-endothelial adherence occurred during hypoxia in rats acclimatized to hypobaric hypoxia. The results are consistent with increased nitric oxide formation due to expression of inducible nitric oxide synthase during the acclimatization period. Further studies are needed to establish the cause of nitric oxide depletion during acute hypoxia as well as to define the compensatory responses that attenuate hypoxia-induced leukocyte-endothelial adherence in the microvasculature of acclimatized rats.

Acclimatization↗

Hypercholesterolemia promotes P-selectin-dependent platelet-endothelial cell adhesion in postcapillary venules.

OBJECTIVE: The objectives of this study were to determine whether hypercholesterolemia promotes platelet-endothelial cell (P/E) adhesion in murine postcapillary venules and define the contributions of endothelial or platelet associated P-selection to hypercholesterolemia-induced P/E interactions. METHODS AND RESULTS: Wild-type (WT) or P-selectin deficient (P-sel-/-) platelets were isolated and labeled with the fluorochrome CFSE and administered to either WT or P-sel-/- mice placed on a normal diet (ND) or high cholesterol diet (HCD). Intravital videomicroscopy was used to quantify platelet saltation and firm adhesion. HCD-WT mice exhibited a time-dependent increase in P/E cell interactions (relative to ND-WT). Flow cytometry revealed an increased expression of P-selectin on circulating platelets of HCD-WT mice at 2 weeks compared with ND-WT mice. When WT platelets were monitored in HCD-P-sel-/- mice, P/E adhesion was dramatically reduced. However, when P-sel-/- platelets were monitored in HCD-WT recipients, P/E adhesive interactions were reduced even further, comparable to ND-WT mice. CONCLUSIONS: These results indicate that elevated cholesterol levels promote P/E adhesion in postcapillary venules and that whereas both endothelial and platelet P-selectin contribute to hypercholesterolemia-induced recruitment of platelets, platelet-associated P-selectin seems to play a more important role in producing the prothrombogenic phenotype in venules.

Animals↗

Endogenous nitric oxide and prostaglandins synergistically counteract thromboembolism in arterioles but not in venules.

It has been shown that NO and prostacyclin (prostaglandin I(2)) from cultured endothelium synergistically inhibit blood platelet aggregation in vitro. However, it is unknown whether this synergism is also effective in the inhibition of thromboembolism in vivo and, if it is, whether it differs between vessel types. Therefore, the effect of endogenous NO and prostacyclin, in combination or alone, on thromboembolism was studied in an in vivo model. Thromboembolism was induced by micropipette puncture of rabbit mesenteric arterioles and venules (diameter 18 to 40 micrometer). In addition, the influence of wall shear rate was analyzed. In arterioles, the combined inhibition of NO synthase (N(G)-nitro-L-arginine [L-NA] 0.1 mmol/L; local superfusion) and of cyclooxygenase (aspirin [ASA] 100 mg/kg IV) resulted in a pronounced, significant prolongation of embolization duration (median >600 seconds) compared with control (median 153 seconds) or treatment with either L-NA (234 seconds) or ASA (314 seconds). This combined effect of L-NA+ASA was greater than the sum of the individual effects of L-NA and ASA. In contrast, in venules L-NA+ASA had no additional effect on embolization duration (209 seconds) compared with the effect of L-NA alone (230 seconds); ASA alone had no effect (122 seconds; control 72 seconds). Interestingly, only in the L-NA+ASA arterioles did embolization correlate positively with wall shear rate (r(s)=0.687; P=0.028). In conclusion, this study indicates that in arterioles, but not in venules, endogenous NO and prostaglandins synergistically counteract ongoing thromboembolism after vessel wall injury and that the combination of endogenous NO and prostaglandins appears to protect against enhancement of arteriolar thromboembolism by wall shear rate.

Animals↗

Angiotensin II type-1 receptor antagonism attenuates the inflammatory and thrombogenic responses to hypercholesterolemia in venules.

Hypercholesterolemia elicits an inflammatory response in the microvasculature that is accompanied by an increased expression of angiotensin II type-1 receptors (AT1-R) on platelets, leukocytes, and endothelial cells. AT1-R blockade attenuates inflammatory responses to angiotensin II (eg, adhesion molecule expression and reactive oxygen species production). We investigated whether AT1-R antagonism attenuates the platelet and leukocyte recruitment induced by acute hypercholesterolemia in postcapillary venules. Leukocyte and platelet adhesion and oxidative stress were quantified by intravital microscopy in cremaster muscle, and P-selectin and AT1-R expression was determined in mice placed on a normal diet (ND) or high-cholesterol diet (HCD) for 2 weeks. Platelet and leukocyte adhesion was significantly elevated by hypercholesterolemia. In HCD mice receiving losartan (HCD-Los) in drinking water, platelet and leukocyte recruitment was reduced to ND levels. Increased platelet adhesion was observed in HCD mice receiving platelets from HCD-Los mice, consistent with a direct beneficial action of losartan on the vessel wall. Hypercholesterolemia elicited an oxidative stress in venules and an increased expression of P-selectin and AT1-R. The oxidative stress and AT1-R upregulation were reduced by losartan, but the P-selectin response was not. We propose that AT1-R engagement contributes to the prothrombogenic and proinflammatory state induced in venules by hypercholesterolemia.

Angiotensin II Type 1 Receptor Blockers↗

Near-infrared spectrophotoscopy of finger venules in assessment of autonomic dysfunction.

The authors evaluated morphologic changes in the venules of the finger using near-infrared spectrophotoscopy in patients with autonomic dysfunction, such as familial amyloidotic polyneuropathy and multiple-system atrophy. Abnormalities of the venules, such as tortuosity, irregular venous caliber, and microaneurysm-like change, and a linear negative correlation between the degree of orthostatic hypotension and the degree of vasoconstriction of the venules were observed.

Adult↗

Microvascular blood is distributed more to venules than to arterioles in patients with Buerger's disease. Observation of bulbar conjunctiva by intravital microscope system.

BACKGROUND: It has been a matter of controversy whether abnormalities of organs other than extremities may be a clinical manifestation of Buerger's disease (thromboangiitis obliterans; TAO). In the present investigation, our aim was to quantitatively characterise the configuration of microvascular networks in bulbar conjunctiva, which is not affected apparently, in patients with thromboangiitis obliterans. METHODS: Nine men with thromboangiitis obliterans attended our hospital and nine male volunteers as normal controls were enrolled in this study. We observed and analysed the configuration of the network of a bulbar conjunctiva by use of intravital microscope system with computer assisted image processing functions. Microvessel density was defined as a summation of vessel length in a ROI area and tortuosity was evaluated by a ratio of vessel length to direct distance of both terminals. RESULTS: In the microcirculation of bulbar conjunctiva in thromboangiitis obliterans, arteriole diameter was significantly decreased and density of venules was significantly increased. Increased venular density was mainly explained by increased tortuosity of venules. CONCLUSIONS: Consequently, microvascular blood was distributed more to venules than to arterioles in patients with thromboangiitis obliterans. Venule/arteriole ratios of diameter, tortuosity and microvessel density may be useful parameters to characterize the configuration of microvascular networks in thromboangiitis obliterans.

Adult↗

[Characteristics of lymphocyte recirculation through postcapillary venules of lymph node paracortical zone following antigenic stimulation].

The walls of venules of paracortical zone of popliteal lymph nodes were studied in 127 2-5 Ys old male dogs (15-20 kg of body weight) after subcutaneous injection of BCG vaccine 0.2 mg/kg in dose. 6 hrs after the administration of BCG vaccine the venule diameter was found to reduce. Then it gradually increased and returned to initial parameters after 3 months. Lymphocyte number was increased in venular lumen and walls following antigenic treatment. Microvilli number growth was observed in endotheliocytes. Basement membrane in the venules was loosened. Lymphocytes through endotheliocytes. Maximal lymphocyte activity and modifications of venular walls was observed on d 7 following the BCG vaccine administration.

Animals↗

Effect of Panax notoginseng saponins on lipopolysaccharide-induced adhesion of leukocytes in rat mesenteric venules.

Panax notoginseng is the root of the Chinese traditional herb, Panax notoginseng (Burk) F.H. Chen. This study was aimed to investigate the inhibitory effect of Panax notoginseng saponins (PNS) on the leukocyte adhesion and the expression of adhesion molecules in rat mesentery venules. Male Sprague-Dawley rats were anesthetized with urethane. These were divided into control, LPS (perfused with lipopolysaccharide), and PNS group (perfused with PNS). The mesenteric microcirculation was observed under a videomicroscope. The number of adherent leukocytes, which attached to the vascular wall during more than 10 seconds, was counted along single venules (30-50 microm in diameter, 200 microm in length). The expression of adhesion molecules was examined using flow-cytometry in blood which was taken from the abdominal aorta and incubated with FITC-labeled CD11b (or CD18) antibodies. The results showed that different changes in the leukocyte adhesion and the expression of adhesion molecules among three groups. In LPS group, the leukocyte adhesion increased significantly after 20 minutes during the observation time, while it was reduced markedly in PNS group. The expression of CD11b and CD18 on the neutrophils was induced in LPS group, while it was reduced significantly in PNS group. It was suggested that PNS could reduce leukocyte adhesion in venules under the inhibitory effect on the expression of adhesion molecules (CD11b and CD18) on neutrophils.

Animals↗

Lymphocyte migration through the wall of postcapillary venules in the fetal development of human palatine tonsil.

Migration of lymphoid cells through the wall of postcapillary venules was studied in the palatine tonsils of five human fetuses. The direction of the migration of lymphoid cells as observed in the early fetal period seems to be mostly from the lumen of the venule into the tonsillar parenchyma. Migration starts with the adherence of the lymphocyte by means of a cytoplasmic projection to the cellular surface of a high-endothelial cell. The projection slips between two neighbouring endothelial cells and extends to settle on the basal membrane of the endothelium. After the disruption of the basal lamina the lymphocyte migrates into the subendothelial connective tissue and further into the extrafollicular tissue of the tonsil. The possibility of transendothelial migration of lymphoid cell through the wall of the postcapillary venule is discussed. Even in case, when the lymphocyte appears to be completely enveloped by the cytoplasm of the endothelial cell it may be in reality the intercellular migration--the lymphoid cell is pressed into the cytoplasm of an endothelial cell.

Cell Movement↗

Role of lipoxygenase, cyclooxygenase and cytochrome P-450 metabolites in contractions of isolated guinea pig pulmonary venules induced by hypoxia and anoxia.

Arachidonic acid metabolites, notably leukotrienes (LTs), have been postulated to play a role in hypoxic pulmonary vasoconstriction. In the present study, we examined the contribution of arachidonic acid metabolites, via the cyclooxygenase, 5-lipoxygenase and cytochrome P-450 monooxygenase pathways, to the hypoxia (25 +/- 3 torr)- and anoxia (0 +/- 2 torr)-induced contractions of isolated pulmonary venules. Neither the cyclooxygenase inhibitors indomethacin (5 microM) or ibuprofen (10 microM) nor the 5-lipoxygenase inhibitors nordihydroguaiaretic acid (5 microM) or U 60257B (10 microM) affected the contractile responses. Similarly, the LT receptor antagonists FPL 57231 (3 microM) or LY 163443 (1 microM), at concentrations that inhibited LT-induced venular contractions, did not significantly affect the responses to hypoxia or anoxia. In fact, anoxia suppressed spontaneous LT release from the venules. The cytochrome P-450 inhibitor SKF-525A (500 microM) nonselectively depressed venular contractions induced by decreased PO2 and pharmacological agents. Induction of the cytochrome P-450 monooxygenase system with beta-naphthoflavone did not alter venular contractions induced by hypoxia or anoxia. Contractions of isolated guinea pig pulmonary venules elicited by decreased PO2 are not mediated by 5-lipoxygenase or cyclooxygenase metabolites. Furthermore, the data do not support a role for cytochrome P-450 metabolites of endogenous substrates in these contractions.

Acetophenones↗

Comparative effects of magnesium salts on reactivity of arterioles and venules to constrictor agents: an in situ study on microcirculation.

The influence of different magnesium salts (i.e., MgCl2, MgSO4, Mg aspartate HCl and Mg acetate) on reactivity of rat mesenteric arterioles (10-25 micron I.D.) and venules (15-30 micron I.D.) to standard constrictor doses of epinephrine and BaCl2 was examined utilizing perivascular, i.a. and i.v. administration of each Mg salt. Perivascular application (0.1, 1.0 and 10 mumol) of the four Mg salts on the microvessels attenuated the vasoconstrictor actions of both epinephrine and BaCl2, dose-dependently. With respect to arterioles, the vasoconstriction induced by BaCl2 was inhibited more potently by each Mg salt than that induced by epinephrine; the reverse was observed with respect to the venules. MgCl2 displayed the most potent antagonism to the constriction induced by BaCl2 in arterioles as well as by epinephrine in venules. Infusion of each Mg salt in very low doses (e.g., 0.1 mumol/min i.a.) resulted in significant attenuation of the arteriolar constrictions induced by both stimulants. However, the difference in the degree of attenuation on i.a. infusion of each Mg salt was significantly less than that observed by perivascular pretreatment. Arteriolar constrictions induced by epinephrine and BaCl2 were also attenuated by systemic i.v. infusion of each Mg salt, except for MgSO4, when low doses (e.g., 0.1 and 1.0 mumol/min.) were given; surprisingly, high dose i.v. infusion (10 and 20 mumol/min) often potentiated arteriolar constrictions induced by both stimulants, but the magnitude of this potentiation depended on the Mg salt utilized.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetates↗