SUMMARYIn the developing chicken embryo yolk sac vasculature, the expression of arterial identity genes requires arterial hemodynamic conditions. We hypothesize that arterial flow must provide a unique signal that is relevant for supporting arterial identity gene expression and is absent in veins. We analyzed factors related to flow, pressure and oxygenation in the chicken embryo vitelline vasculature in vivo. The best discrimination between arteries and veins was obtained by calculating the maximal pulsatile increase in shear rate relative to the time-averaged shear rate in the same vessel: the relative pulse slope index (RPSI). RPSI was significantly higher in arteries than veins. Arterial endothelial cells exposed to pulsatile shear in vitro augmented arterial marker expression as compared with exposure to constant shear. The expression of Gja5 correlated with arterial flow patterns: the redistribution of arterial flow provoked by vitelline artery ligation resulted in flow-driven collateral arterial network formation and was associated with increased expression of Gja5. In situ hybridization in normal and ligation embryos confirmed that Gja5 expression is confined to arteries and regulated by flow. In mice, Gja5 (connexin 40) was also expressed in arteries. In the adult, increased flow drives arteriogenesis and the formation of collateral arterial networks in peripheral occlusive diseases. Genetic ablation of Gja5 function in mice resulted in reduced arteriogenesis in two occlusion models. We conclude that pulsatile shear patterns may be central for supporting arterial identity, and that arterial Gja5 expression plays a functional role in flow-driven arteriogenesis.

A theoretical method is used to simulate the motion and deformation of mammalian red blood cells (RBCs) in microvessels, based on knowledge of the mechanical characteristics of RBCs. Each RBC is represented as a set of interconnected viscoelastic elements in two dimensions. The motion and deformation of the cell and the motion of the surrounding fluid are computed using a finite-element numerical method. Simulations of RBC motion in simple shear flow of a high-viscosity fluid show "tank-treading'' motion of the membrane around the cell perimeter, as observed experimentally. With appropriate choice of the parameters representing RBC mechanical properties, the tank-treading frequency and cell elongation agree closely with observations over a range of shear rates. In simulations of RBC motion in capillary-sized channels, initially circular cell shapes rapidly approach shapes typical of those seen experimentally in capillaries, convex in front and concave at the rear. An isolated RBC entering an 8-mum capillary close to the wall is predicted to migrate in the lateral direction as it traverses the capillary, achieving a position near the center-line after traveling a distance of about 60 mum. Cell trajectories agree closely with those observed in microvessels of the rat mesentery.

Adaptation of vascular networks to functional demands needs vessel growth, vessel regression and vascular remodelling. Biomechanical forces resulting from blood flow play a key role in these processes. It is well-known that metabolic stimuli, mechanical forces and flow patterns can affect gene expression and remodelling of vascular networks in different ways. For instance, in the sprouting type of angiogenesis related to hypoxia, there is no blood flow in the rising capillary sprout. In contrast, it has been shown that an increase of wall shear stress initiates the splitting type of angiogenesis in skeletal muscle. Otherwise, during development, both sprouting and intussusception act in parallel in building the vascular network, although with differences in spatiotemporal distribution. Thereby, in addition to regulatory molecules, flow dynamics support the patterning and remodelling of the rising vascular tree. Herewith, we present an overview of angiogenic processes with respect to intussusceptive angiogenesis as related to local haemodynamics.

We report that adhesion-induced eosinophil cytolysis takes place through RIPK3-MLKL-dependent necroptosis, which can be counterregulated by autophagy.