Abstract:Sprouting angiogenesis is a well-coordinated process controlled by multiple extracellular inputs, including vascular endothelial growth factor (VEGF). However, little is known about when and how individual endothelial cell (EC) responds to angiogenic inputs in vivo. Here, we visualized endothelial Ca2+ dynamics in zebrafish and found that intracellular Ca2+ oscillations occurred in ECs exhibiting angiogenic behavior. Ca2+ oscillations depended upon VEGF receptor-2 (Vegfr2) and Vegfr3 in ECs budding from the do… Show more
“…Since the body of a zebrafish larva is transparent through all developmental stages, such transgenic fish have been used to visualize the vascular system [108], the central and peripheral nervous systems [109, 110], the regenerating processes of the sensory system and fin [111, 112], the cellular process of muscle wound repair [113], and proteolytic cleavage of the extracellular domain of a membrane protein (Neuregulin) in the motor neurons [114], among others. Also, the transgenic fish were applied to image the activity of specific neuronal populations or endothelial cells by targeted expression of a calcium indicator GCaMP [115, 116]. In addition, gene trap vectors similar to those shown in Fig.…”
Section: Transposons and Functional Genomicsmentioning
Genetic tools and mutagenesis strategies based on transposable elements are currently under development with a vision to link primary DNA sequence information to gene functions in vertebrate models. By virtue of their inherent capacity to insert into DNA, transposons can be developed into powerful tools for chromosomal manipulations. Transposon-based forward mutagenesis screens have numerous advantages including high throughput, easy identification of mutated alleles and providing insight into genetic networks and pathways based on phenotypes. For example, the Sleeping Beauty transposon has become highly instrumental to induce tumors in experimental animals in a tissue-specific manner with the aim of uncovering the genetic basis of diverse cancers. Here, we describe a battery of mutagenic cassettes that can be applied in conjunction with transposon vectors to mutagenize genes, and highlight versatile experimental strategies for the generation of engineered chromosomes for loss-of-function as well as gain-of-function mutagenesis for functional gene annotation in vertebrate models, including zebrafish, mice and rats.
“…Since the body of a zebrafish larva is transparent through all developmental stages, such transgenic fish have been used to visualize the vascular system [108], the central and peripheral nervous systems [109, 110], the regenerating processes of the sensory system and fin [111, 112], the cellular process of muscle wound repair [113], and proteolytic cleavage of the extracellular domain of a membrane protein (Neuregulin) in the motor neurons [114], among others. Also, the transgenic fish were applied to image the activity of specific neuronal populations or endothelial cells by targeted expression of a calcium indicator GCaMP [115, 116]. In addition, gene trap vectors similar to those shown in Fig.…”
Section: Transposons and Functional Genomicsmentioning
Genetic tools and mutagenesis strategies based on transposable elements are currently under development with a vision to link primary DNA sequence information to gene functions in vertebrate models. By virtue of their inherent capacity to insert into DNA, transposons can be developed into powerful tools for chromosomal manipulations. Transposon-based forward mutagenesis screens have numerous advantages including high throughput, easy identification of mutated alleles and providing insight into genetic networks and pathways based on phenotypes. For example, the Sleeping Beauty transposon has become highly instrumental to induce tumors in experimental animals in a tissue-specific manner with the aim of uncovering the genetic basis of diverse cancers. Here, we describe a battery of mutagenic cassettes that can be applied in conjunction with transposon vectors to mutagenize genes, and highlight versatile experimental strategies for the generation of engineered chromosomes for loss-of-function as well as gain-of-function mutagenesis for functional gene annotation in vertebrate models, including zebrafish, mice and rats.
“…Following the ‘decide then move’ time-ordering, rearrangements should presumably be quite regular, with the timing driven by gene expression delays in the Hes1 oscillator. In a related in vivo zebrafish study, Yokota et al [61] asked the temporal question ‘ When do ECs respond to VEGF?’ providing a tantalizing peak into exactly these kind of early selection dynamics. By utilizing the fast frame rates of lightsheet microscopy they live-imaged Ca 2+ oscillations found to occur in VEGF–VEGFR-2 activated ECs during zebrafish intersegmental vessel (ISV) growth.…”
Section: A Novel Time-based Formulation Of Endothelial Cell Competitimentioning
confidence: 99%
“…Finally, the topology of the live-imaged lung explant model was quantified and found to be significantly less branched. It is interesting to note that Sema3E-Plexin-D1 signalling as the intersection between somites and the DA was found to affect the collective dynamics of EC activation [61]. Figure 4.Sema3E-PlexinD1 speeds up the CPG increasing branching density ( a ) pathway schematic showing crosstalk with the CPG.…”
Section: Changing the Tempo To Alter Vascular Structurementioning
The process of new blood vessel growth (angiogenesis) is highly dynamic, involving complex coordination of multiple cell types. Though the process must carefully unfold over time to generate functional, well-adapted branching networks, we seldom hear about the time-based properties of angiogenesis, despite timing being central to other areas of biology. Here, we present a novel, time-based formulation of endothelial cell behaviour during angiogenesis and discuss a flurry of our recent, integrated in silico/in vivo studies, put in context to the wider literature, which demonstrate that tissue conditions can locally adapt the timing of collective cell behaviours/decisions to grow different vascular network architectures. A growing array of seemingly unrelated ‘temporal regulators’ have recently been uncovered, including tissue derived factors (e.g. semaphorins or the high levels of VEGF found in cancer) and cellular processes (e.g. asymmetric cell division or filopodia extension) that act to alter the speed of cellular decisions to migrate. We will argue that ‘temporal adaptation’ provides a novel account of organ/disease-specific vascular morphology and reveals ‘timing’ as a new target for therapeutics. We therefore propose and explain a conceptual shift towards a ‘temporal adaptation’ perspective in vascular biology, and indeed other areas of biology where timing remains elusive.This article is part of the themed issue ‘Systems morphodynamics: understanding the development of tissue hardware’.
“…ECs in the segmental vessels induces intracellular calcium oscillations and these oscillations are important in the determination of the tip cells (Yokota et al, 2015). The growing ISVs follow the intersomitic fissure until they reach the myoseptum, from which the tip cells migrate towards the dorsal roof of the neural tube (Ellertsdóttir et al, 2010).…”
Section: Tip Cell Selection and Functionmentioning
Vascular networks are formed and maintained through a multitude of angiogenic processes, such as sprouting, anastomosis and pruning. Only recently has it become possible to study the behavior of the endothelial cells that contribute to these networks at a single-cell level in vivo. This Review summarizes what is known about endothelial cell behavior during developmental angiogenesis, focusing on the morphogenetic changes that these cells undergo.
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