The effect of absent blood flow on the zebrafish cerebral and trunk vasculature
Posted on: 3 September 2020
Preprint posted on 24 July 2020
Article now published in Vascular Biology at http://dx.doi.org/10.1530/vb-21-0009
Microscopy and zebrafish: Understanding blood-flow and its effects on different vascular beds – part I.
Selected by Mariana De NizCategories: cell biology, developmental biology
Background
Endothelial cells perform crucial functions in health and disease. They are important for physiological processes including wound healing, tissue regeneration, immune responses, menstruation and pregnancy. Increasing evidence suggests endothelial cells display different molecular and functional properties according to their anatomical site, emphasising the need to study endothelial cell function in different vascular beds. Zebrafish are a conventionally used model organism to study the vasculature in health and disease.|Zebrafish embryonic transparency allows non-invasive studies of different vascular beds in the same animal, and zebrafish embryos are well-established models to study the role of blood flow on vasculogenesis and angiogenesis.
Important questions that remain unanswered about the role of blood flow in vascular development include whether the absence of blood flow induces similar effects in different vascular beds, and what that effect is over time. In their work, Kugler et al performed LSFM 3D in vivo imaging to study the head and trunk vasculature of zebrafish embryos with and without blood flow (1).
Key findings and developments
The study by Kugler et al begins by determining whether cerebrovascular patterning is impaired by absent blood flow. The authors began by comparing the cerebral vascular morphology of controls, control morpholinos, and troponin T2A morpholinos that have inhibited development of cardiac contraction and impeded blood flow. The latter displayed abnormal cerebral vasculature, with vessels in the midbrain severely affected, and an enlarged primary head sinus, while the perineural vessels remained relatively normal. Examination of the distribution of endothelial cells and nuclear density in these animals supported these findings. Altogether, the main finding was that the effects of lack of blood flow become more severe over time, and that central vessels are more impacted than those in the periphery.
The study continued by examining vascular patterning and endothelial cell nuclei in the trunk of the 3 groups of animals previously established. In contrast to cerebral vessels, overall, trunk vessel patterning seemed unaltered in the absence of blood flow, except for the cardinal vein which seemed affected by lack of blood flow as remodelling, including intussusception – the creation of new blood vessels created by splitting of an existing blood vessel into two, seemed lacking confirming previous findings by (Karthik et al., 2018 (2)).
The authors went on to elucidate the effect of absent blood flow over time by quantifying cerebrovascular area and number. They found a significant reduction in vascular area and endothelial cell number in troponin T2A morphants. During development, the number of nuclei-to-vasculature increased in controls, but decreased in animals without blood flow. A similar analysis in the trunk showed that the vascular area was significantly reduced by absent blood flow early in embryo development, but not later. Altogether, these results suggested that the cerebral vessels were more severely affected than the trunk vessels.
To determine whether vessels of different identity were differentially impacted by lack of blood flow, the diameter of different veins and arteries in the brain, trunk and tail were quantified. Cerebral arterial diameter was reduced by 2.5% in troponin T2A morphants compared to controls, while posterior cerebral vein diameter was reduced by 36%. In the trunk, the mean arterial diameter was reduced by 40%, while the venous diameter was reduced by 46%. Finally, the dorsal aorta was reduced by 41%, while the posterior cardinal vein was reduced by 29%. These results suggest that vessels of arterial and venous identity are both affected by lack of blood flow, but specific differences exist in various anatomic locations.
The study then went on to explore whether cell death contributed to a reduction in endothelial cell numbers in the troponin T2A morphants. In the head and trunk, cell death appeared to be increased in the morphants, however more detailed analysis suggested the increased signal of the cell death marker came from non-vascular foci, showing that increased cell death occurs in the absence of blood flow, but is not endothelial-cell specific.
The authors hypothesized that perhaps the increased cell death in the troponin T2A morphants might alter the inflammatory response present in different anatomical locations. However, no difference was found in the number of neutrophils or macrophages between the three zebrafish groups used in the study. Nitric oxide, which was used as indirect inflammation read-out, also did not appear to differ in the head or trunk. Altogether, the data suggest that the lack of blood flow increases cell death, but not tissue inflammation.
What I like about this preprint
I find vascular biology very interesting and I think the authors used an exciting imaging method and an exciting model to answer questions on vascular biology, relevant to various field of research.
References
- Kugler et al, The effect of absent blood flow on the zebrafish cerebral and trunk vasculature, bioRxiv, 2020.
-
Karthik, S., et al., Synergistic interaction of sprouting and intussusceptive angiogenesis during zebrafish caudal vein plexus development. Scientific Reports 8, 2018.
doi: https://doi.org/10.1242/prelights.24413
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