Effects of erythrocyte aggregation and venous network geometry on red blood cell axial migration.

Axial migration of red blood cells in small glass tubes can cause blood viscosity to be effectively independent of shear rate. However, this phase separation may not occur to the same degree in the venous network due to infusion of cells and aggregates at branch points. To investigate this hypothesis, we followed trajectories of fluorescently labeled red blood cells in the venular network of the rat spinotrapezius muscle at normal and reduced flow with and without red blood cell aggregation. Cells traveling near the wall of an unbranched venular segment migrated approximately 1% of the longitudinal path length without aggregation and migrated slightly more with aggregation. Venular segment length between branch points averaged three to five times the diameter. Cells in the main vessel were shifted centrally by up to 20% of diameter at branch points, reducing the migration rate of cells near the opposite wall to <1% even in the presence of aggregation. We conclude that formation of a cell-free marginal layer in the venular network is attenuated due to the time dependence of axial migration and the frequent branching of the network.

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.

Rheological effects of red blood cell aggregation in the venous network: a review of recent studies.

It has long been recognized that understanding the rheological properties of blood is essential to a full understanding of the function of the circulatory system. Given the difficulty of obtaining carefully controlled measurements in vivo, most of our current concepts of the flow behavior of blood in vivo are based on its properties in vitro. Studies of blood rheology in rotational and tube viscometers have defined the basic properties of blood and pointed to certain features that may be especially significant for understanding in vivo function. At the same time, differences between in vivo and in vitro systems combined with the complex rheological properties of blood make it difficult to predict in vivo blood rheology from in vitro studies. We have investigated certain flow properties of blood in vivo, using the venular network of skeletal muscle as our model system. In the presence of red blood cell aggregation, venous velocity profiles become blunted from the parabolic as in Poiseuille flow, as pseudo-shear rate (= mean fluid velocity/vessel diameter) is decreased from approximately 100 s(-1) to 5 s(-1). At control flow rates, the short distance between venular junctions does not appear to permit significant axial migration and red cell depletion of the peripheral fluid layer before additional red cells and aggregates are infused from a feeding tributary. Formation of a cell-free plasma layer at the vessel wall and sedimentation in vivo are evident only at very low pseudo-shear rates (<5 s(-1)). These findings may explain in large part observations in whole organs of increased venous resistance with reduction of blood flow.

Effect of erythrocyte aggregation on velocity profiles in venules.

A recent whole organ study in cat skeletal muscle showed that the increase in venous resistance seen at reduced arterial pressures is nearly abolished when the muscle is perfused with a nonaggregating red blood cell suspension. To explore a possible underlying mechanism, we tested the hypothesis that red blood cell aggregation alters flow patterns in vivo and leads to blunted red blood cell velocity profiles at reduced shear rates. With the use of fluorescently labeled red blood cells in tracer quantities and a video system equipped with a gated image intensifier, we obtained velocity profiles in venous microvessels (45-75 microm) of rat spinotrapezius muscle at centerline velocities between 0.3 and 14 mm/s (pseudoshear rates 3-120 s(-1)) under normal (nonaggregating) conditions and after induction of red blood cell aggregation with Dextran 500. Profiles are nearly parabolic (Poiseuille flow) over this flow rate range in the absence of aggregation. When aggregation is present, profiles are parabolic at high shear rates and become significantly blunted at pseudoshear rates of 40 s(-1) and below. These results indicate a possible mechanism for increased venous resistance at reduced flows.

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%.

A High-Affinity Hemoglobin Is Expressed in the Notochord of Amphioxus, Branchiostoma californiense.

In the phylum Chordata, only members of the subphylum Vertebrata were thought to express hemoglobin (Hb). Here we document the existence of intracellular Hb expressed in members of the subphylum Cephalochordata. Hemoglobin is expressed in myotome tissue and in notochord cells within the body of amphioxus. Both notochord and myotome tissue Hbs have a molecular size consistent with a dimeric molecule made up of two non-covalently linked monomers each of approximately 19 kD. The notochord Hb has a relatively high oxygen-binding affinity, with an apparent P5O of 0.036 kPa (0.27mm Hg), and it does not bind oxygen cooperatively. The notochord Hb may be involved in facilitating oxygen delivery and providing a short-term oxygen store within the notochord cells in order to maintain a high level of aerobic metabolism in support of the sustained contraction necessary for notochord function.