Novel mathematical morphology model identifies dorsal-ventral asymmetry of endothelial cell morphology in dorsal aorta of wild-type and Endoglin-deficient zebrafish embryos

Abstract

Endothelial cells, which line the lumen of blood vessels, locally sense and respond to blood flow. In response to altered blood flow dynamics during early embryonic development, these cells undergo shape changes that directly affect vessel geometry: In the dorsal aorta of zebrafish embryos, elongation of endothelial cells in the direction of flow between 48 and 72 hours post fertilization (hpf) reduces the vessels diameter. This remodeling process requires Endoglin; excessive endothelial cell growth in the proteins absence results in vessel diameter increases. To understand how these changes in vessel geometry emerge from morphological changes of individual endothelial cells, we developed a novel mathematical model of the dorsal aortas apico-luminal surface that allows simultaneous quantification of vessel geometry and endothelial cell morphology. Based on fluorescently marked endothelial cell contours, we inferred cross-sections of the dorsal aorta that accounted for dorsal flattening of the vessel. By projection of endothelial cell contours onto the estimated cross-sections and subsequent triangulation, we finally reconstructed 3D surfaces of the individual cells. By simultaneously reconstructing vessel cross-sections and cell surfaces, we found that cell morphology varied between endothelial cells located in different sectors of the dorsal aorta in both wild-type and Endoglin-deficient zebrafish embryos: In wild-types, ventral endothelial cells were smaller and more elongated in flow direction than dorsal endothelial cells at both 48 hpf and 72 hpf. Although dorsal and ventral endothelial cells in Endoglin-deficient embryos had similar sizes at 48 hpf, dorsal endothelial cells were much larger at 72 hpf. In Endoglin-deficient embryos, elongation in flow direction increased between 48 hpf and 72 hpf in ventral endothelial cells but hardly changed in dorsal endothelial cells. Hereby, we provide evidence that dorsal endothelial cells contribute most to the disparate changes in dorsal aorta diameter in wild-type and Endoglin-deficient embryos between 48 hpf and 72 hpf. Author summaryEndothelial cells, which form the innermost layer of each blood vessel, sense and respond to blood flow. During early embryonic development in zebrafish, endothelial cells of the dorsal aorta elongate in the direction of blood flow and hereby decrease the vessels diameter. To understand how these changes in vessel geometry emerge from morphological changes of individual endothelial cells, it is critical to precisely quantify both vessel geometry and cell morphology. To this end, we developed a 3D mathematical model of the dorsal aorta. Leveraging information from fluorescently marked endothelial cell contours allowed us to reconstruct the vessels surface. We applied this method to wild-type and mutant zebrafish embryos lacking functional Endoglin that is required for the physiological vessel diameter decrease. By quantifying vessel geometry and cell morphology in these embryos, we found that cell size and elongation in the direction of blood flow varied between endothelial cells located in different vessel sectors. Notably, we determined a subgroup of endothelial cells that contributed most to the vessel diameter increases in the absence of Endoglin. Future studies can investigate whether variability in endothelial cell behavior also contributes to the onset of human vascular malformations occurring due to a loss of Endoglin.