Biology subjects

Kotini, M. P.

Publications and source records attributed to Kotini, M. P..

5 recordsLinked to original sources

bioRxiv2025

An AmotL2 - Yap1 Module Integrates Flow and Junctional Mechanics to Specify Vascular Pruning Hotspots

Vascular networks constantly remodel to optimise blood flow, yet how endothelial cells (ECs) integrate haemodynamic cues with junctional mechanics to execute precise branch pruning remains unclear. Using live imaging and quantitative topology analysis in the zebrafish sub-intestinal vein plexus, we define the cellular events underlying flow-guided regression. Pruning occurs in geometrically disadvantaged, low-flow branches and is executed through transient polarity fluctuations, junctional remodelling and directed EC migration. We identify AmotL2 as an essential regulator of endothelial mechanical competence. AmotL2 maintains junctional architecture, actomyosin tension and appropriate cell packing, enabling ECs to polarise and rearrange in response to local flow differences. Loss of AmotL2 disrupts tension homeostasis, increases tissue compaction and prevents branch regression despite normal perfusion. In contrast, Yap1 plays an opposing, stabilising role, and its loss results in excessive pruning and network simplification. Together, AmotL2 and Yap1 constitute a mechanosensitive balancing module that interprets flow magnitude and vascular geometry to determine branch stability. This framework describes how mechanical forces sculpt vascular architecture during development and homeostasis.

developmental biology↗

bioRxiv2024

Initiation of Lumen Formation from Junctions via Differential Actomyosin Contractility Regulated by Dynamic Recruitment of Rasip1

Vascular lumens are essential for the dispersal of nutrients, oxygen, and immune cells throughout the body. De novo lumen formation necessitates the precise segregation of junctional proteins from apical surfaces. Direct visualization of this process and a comprehensive understanding on how the apical and junctional compartments are segregated and established remain elusive, due to the lack of visualizable systems, proper reporters and manipulative tools. Using zebrafish as an experimental model, we have established a wealth of molecular reporters, photo-convertible and optogenetic tools and combined them with genetic analyses to study the establishment of apical domains from pre-apical junctional complexes. Our study identified Rasip1 as one of the earliest apical proteins recruited, peripheralizing cortical contractility at the junctional patches by inhibiting NMII at the center, thereby initiating the outward flow of junctional complexes. After the establishment of the apical compartments, Rasip1 shuttles between junctions and the apical compartments to the high-tension sites and tunes down the local hyper contractility, restricting Cdh5 to junctions by suppressing apical contractility. Above all, the formation of de novo lumens and maintenance of junctional integrity depend on the precise control of contractility within the apical compartments and junctions, orchestrated by the dynamic recruitment of Rasip1.

developmental biology↗

bioRxiv2023

Genetic analysis of rab7 mutants in zebrafish

Vascular network formation requires the fusion of newly formed blood vessels and the emergence of a patent lumen between the newly established connections so that blood flow can start. Lumen formation has been shown to depend on the late endosomal/lysosomal pathway in various organs of animal tubular systems. Here, we identified a late endosomal/lysosomal vesicular fraction (Rab7/Lamp2) in early zebrafish angiogenic sprouts, which appears to contribute to apical membrane growth during lumen formation. To study the effect of the late endocytic pathway on vascular development, we generated mutant alleles for all three rab7 genes in zebrafish (rab7a, rab7ba, rab7bb). All rab7 genes are expressed in wild-type zebrafish and we did not detect any compensatory effects by the other rab7 isoforms in single knockout mutants, which were all viable. Only the triple mutant was lethal suggesting some functional redundancy. However, the different rab7 isoforms fulfil also at least partially independent functions because eggs laid from mothers lacking two rab7 (rab7a and/or rab7bb). showed reduced survival and contained enlarged yolk granules, suggesting maternal contribution of these two rab7. Finally, we observed minor effects on lumen formation in embryos which still express one copy of rab7. Our results support the notion that the late endocytic/lysosomal compartment contributes to lumen expansion.

genetics↗

bioRxiv2021

Vinculin controls endothelial cell junction dynamics during vascular lumen formation

Blood vessel morphogenesis is driven by coordinated endothelial cell behaviors, which depend on dynamic cell-cell interactions. Remodeling of endothelial cell-cell junctions promote morphogenetic cellular events while preserving vascular integrity. Here, we have analyzed the dynamics of endothelial cell-cell junctions during lumen formation in angiogenic sprouts. By live-imaging of the formation of intersegmental blood vessels in zebrafish, we demonstrate that lumen expansion is accompanied by the formation of transient finger-shaped junctions. Formation and maintenance of these junctional fingers are positively regulated by blood pressure whereas inhibition of blood flow prevents their formation. Using fluorescent reporters, we show that the tension-sensor Vinculin localizes to junctional fingers. Furthermore, loss of vinculin function, in vinculin a and -b double knockouts, prevents junctional finger formation in angiogenic sprouts, whereas endothelial expression of a vinculin transgene is sufficient to restore junctional fingers. Taken together, our findings suggest a mechanism in which lumen expansion during angiogenesis leads to an increase in junctional tension, which triggers recruitment of vinculin and formation of junctional fingers. We propose that endothelial cells may employ force-dependent junctional remodeling to react to changes in external forces to protect cell-cell contacts and to maintain vascular integrity during sprouting angiogenesis.

cell biology↗

bioRxiv2018

Sprouting and anastomosis in the Drosophila trachea and the vertebrate vasculature: similarities and differences in cell behaviour

Branching morphogenesis is a fascinating process whereby a simple network of biological tubes increases its complexity by adding new branches to existing ones, generating an enlarged structure of interconnected tubes. Branching morphogenesis has been studied extensively in animals and much has been learned about the regulation of branching at the cellular and molecular level. Here, we discuss studies of the Drosophila trachea and of the vertebrate vasculature, which have revealed how new branches are formed and connect (anastomose), leading to the establishment of complex tubular networks. We briefly describe the cell behaviour underlying tracheal and vascular branching. Although similar at many levels, the branching and anastomosis processes characterized thus far show a number of differences in cell behaviour, resulting in somewhat different tube architectures in these two organs. We describe the similarities and the differences and discuss them in the context of their possible developmental significance. We finish by highlighting some old and new data, which suggest that live imaging of the development of capillary beds in adult animals might reveal yet unexplored endothelial behaviour of endothelial cells.\n\nO_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=161 SRC=\"FIGDIR/small/458281_ufig1.gif\" ALT=\"Figure 1\">\nView larger version (25K):\norg.highwire.dtl.DTLVardef@1f61f04org.highwire.dtl.DTLVardef@16e76fborg.highwire.dtl.DTLVardef@1458fd7org.highwire.dtl.DTLVardef@1e67d51_HPS_FORMAT_FIGEXP M_FIG C_FIG

developmental biology↗