scholarly journals Control of dynamic cell behaviors during angiogenesis and anastomosis by Rasip 1

2020 ◽  
Author(s):  
Minkyoung Lee ◽  
Charles Betz ◽  
Ilkka Paatero ◽  
Niels Schellinx ◽  
Jianmin Yin ◽  
...  
Keyword(s):  
Time Lapse ◽  
Tube Formation ◽  
Interacting Protein ◽  
Cell Behaviors ◽  
Sprouting Angiogenesis ◽  
Phenotype Analysis ◽  
Membrane Compartment ◽  
Dynamic Cell ◽  
Cellular Behaviors ◽  
And Function

AbstractOrgan morphogenesis is driven by a wealth of tightly orchestrated cellular behaviors, which ensure proper organ assembly and function. Many of these cell activities involve cell-cell interactions and remodeling of the F-actin cytoskeleton. Here, we analyze the requirement for Rasip1 (Ras-interacting protein 1), an endothelial-specific regulator of junctional dynamics, during blood vessel formation. Phenotype analysis of rasip1 mutants in zebrafish embryos reveal distinct requirements for Rasip1 during sprouting angiogenesis, vascular anastomosis and lumen formation. During angiogenic sprouting, Rasip1 is required for efficient cell pairing, which is essential for multicellular tube formation. High-resolution time-lapse analyses show that these cell pairing defects are caused by a destabilization of tricellular junctions suggesting that tri-cellular junctions may serve as a counterfort to tether sprouting endothelial cells during morphogenetic cell rearrangements. During anastomosis, Rasip1 is required to establish a stable apical membrane compartment; rasip1 mutants display ectopic, reticulated junctions and the apical compartment is frequently collapsed. Loss of Ccm1 and Heg1 function leads to junctional defects similar to those seen in rasip1 mutants. Analysis of radil-b single and rasip1/radil-b double mutants reveal distinct and overlapping functions of both proteins. While Rasip1 and Radil-b have similar functions during angiogenic sprouting, the junction formation during anastomosis may primarily depend on Rasip1.

scholarly journals Control of dynamic cell behaviors during angiogenesis and anastomosis by Rasip 1

Development ◽  
2021 ◽  
Author(s):  
Minkyoung Lee ◽  
Charles Betz ◽  
Jianmin Yin ◽  
Ilkka Paatero ◽  
Niels Schellinx ◽  
...  
Keyword(s):  
Cell Junctions ◽  
Interacting Protein ◽  
Cell Behaviors ◽  
Sprouting Angiogenesis ◽  
Junction Formation ◽  
Phenotype Analysis ◽  
Membrane Compartment ◽  
Dynamic Cell ◽  
Cellular Behaviors ◽  
And Function

Organ morphogenesis is driven by a wealth of tightly orchestrated cellular behaviors, which ensure proper organ assembly and function. Many of these cell activities involve cell-cell interactions and remodeling of the F-actin cytoskeleton. Here, we analyze the requirement for Rasip1 (Ras-interacting protein 1), an endothelial-specific regulator of junctional dynamics, during blood vessel formation. Phenotype analysis of rasip1 mutants in zebrafish embryos reveal distinct functions of Rasip1 during sprouting angiogenesis, anastomosis and lumen formation. During angiogenic sprouting, loss of Rasip1 causes cell pairing defects due to a destabilization of tricellular junctions, indicating that stable tri-cellular junctions are essential to maintain multicellular organization within the sprout. During anastomosis, Rasip1 is required to establish a stable apical membrane compartment; rasip1 mutants display ectopic, reticulated junctions and the apical compartment is frequently collapsed. Loss of Ccm1 and Heg1 function mimics junctional defects of rasip1 mutants. Furthermore, downregulation of ccm1 and heg1 leads to a delocalization of Rasip1 at cell junctions, indicating that junctional tethering of Rasip1 is required for its function during junction formation and stabilization during sprouting angiogenesis.

scholarly journals Vinculin controls endothelial cell junction dynamics during vascular lumen formation

2021 ◽  
Author(s):  
Maria P. Kotini ◽  
Miesje M. van der Stoel ◽  
Mitchell K. Han ◽  
Bettina Kirchmaier ◽  
Johan de Rooij ◽  
...  
Keyword(s):  
Endothelial Cell ◽  
Cell Junctions ◽  
Cell Junction ◽  
Vascular Integrity ◽  
Lumen Formation ◽  
Cell Behaviors ◽  
Sprouting Angiogenesis ◽  
Dynamic Cell ◽  
External Forces ◽  
Cell Cell

AbstractBlood 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.

scholarly journals Cellular crowding influences extrusion and proliferation to facilitate epithelial tissue repair

2019 ◽  
Vol 30 (16) ◽  
pp. 1890-1899 ◽  
Author(s):  
Jovany J. Franco ◽  
Youmna Atieh ◽  
Chase D. Bryan ◽  
Kristen M. Kwan ◽  
George T. Eisenhoffer
Keyword(s):  
Tissue Repair ◽  
Wound Closure ◽  
Time Lapse ◽  
Cell Number ◽  
Confocal Imaging ◽  
Epithelial Tissue ◽  
Directed Cell Migration ◽  
Epithelial Wound Healing ◽  
Cellular Behaviors ◽  
And Function

Epithelial wound healing requires a complex orchestration of cellular rearrangements and movements to restore tissue architecture and function after injury. While it is well known that mechanical forces can affect tissue morphogenesis and patterning, how the biophysical cues generated after injury influence cellular behaviors during tissue repair is not well understood. Using time-lapse confocal imaging of epithelial tissues in living zebrafish larvae, we provide evidence that localized increases in cellular crowding during wound closure promote the extrusion of nonapoptotic cells via mechanically regulated stretch-activated ion channels (SACs). Directed cell migration toward the injury site promoted rapid changes in cell number and generated shifts in tension at cellular interfaces over long spatial distances. Perturbation of SAC activity resulted in failed extrusion and increased proliferation in crowded areas of the tissue. Together, we conclude that localized cell number plays a key role in dictating cellular behaviors that facilitate wound closure and tissue repair.

scholarly journals ECM cross-linking regulates invadopodia dynamics

2017 ◽  
Author(s):  
Kamyar Esmaeili ◽  
Aviv Bergman ◽  
Bojana Gligorijevic
Keyword(s):  
Cancer Cells ◽  
Cancer Cell ◽  
Time Lapse ◽  
Invasive Cancer ◽  
Cross Linking ◽  
Β1 Integrin ◽  
Cell Behaviors ◽  
Lysine Residues ◽  
Ecm Degradation ◽  
And Function

AbstractInvadopodiInvadopodia are membrane protrusions dynamically assembled by invasive cancer cells in contact with extracellular matrix (ECM). Invadopodia are enriched for the structural proteins actin and cortactin, as well as metalloproteases such as MT1-MMP, whose function is to degrade the surrounding ECM. During metastasis, invadopodia are necessary for cancer cell intravasation and extravasation. While signaling pathways involved in the assembly and function of invadopodia are well studied, few studies address invadopodia dynamics and how the cell-ECM interactions contribute to cell invasion. Using iterative analysis based on time-lapse microscopy and mathematical modeling of invasive cancer cells, we found that cells oscillate between invadopodia presence and cell stasis, termed Invadopodia state and invadopodia absence during cell translocation, termed Migration state. Our data suggests that β1-integrin-ECM binding and ECM cross-linking control the duration of each of the two states. By changing the concentration of cross-linkers in 2D and 3D cultures, we generate ECM where 0-0.92 of total lysine residues are cross-linked. Using ECM with a range of cross-linking degrees we demonstrate that the dynamics of invadopodia-related functions have a biphasic relationship to ECM cross-linking. At intermediate levels of ECM cross-linking (0.39), cells exhibit rapid invadopodia protrusion-retraction cycles and rapid calcium spikes, which lead to more frequent MT1-MMP delivery, causing maximal invadopodia-mediated ECM degradation. In contrast, both extremely high or low levels of cross-linking lead to slower invadopodia-related dynamics and lower ECM degradation. Additionally, β1-integrin inhibition modifies dynamics of invadopodia-related functions, as well as the length of time cells spend in either of the states. Collectively, these data suggest that β1-integrin-ECM binding non-linearly translates small physical differences in extracellular environment to differences in the dynamics of cancer cell behaviors. Understanding conditions under which invadopodia can be reduced by subtle environment-targeting treatments may lead to combination therapies for preventing metastatic spread.

Abstract 250: Haploinsufficiency of Muscle-Enriched A-Type Lamin Interacting Protein (MLIP) Manifests as Enlarged Dilated Hearts

2012 ◽  
Vol 111 (suppl_1) ◽  
Author(s):  
Patrick Burgon ◽  
Julia Lockwood ◽  
Glenn Wells ◽  
Alexandre Blais
Keyword(s):  
Heart Development ◽  
Left Ventricular ◽  
Lamin A ◽  
Interacting Protein ◽  
Cellular Hypertrophy ◽  
Cellular Source ◽  
Alternatively Spliced ◽  
Normal Body Weight ◽  
Heart Size ◽  
And Function

Approximately 116 unique mutations in the lamin A/C gene have been described to date that are associated with dilated cardiomyopathy. We recently reported the discovery of MLIP through its interaction with lamin A/C. MLIP is expressed ubiquitously and most abundantly in heart, skeletal and smooth muscle of amniotes (mammals, reptiles and birds) and has no paralogous homologue suggesting no functional redundancy. The MLIP gene encodes at least seven, alternatively spliced, LMNA-interacting factors that possess several structural motifs not found in any other protein. The MLIP isoforms pattern of expression differs between each of the tissues with heart being the most heterogeneous. Down-regulation of lamin A/C expression by shRNA results in the up-regulation and mis-localization of MLIP. In addition to interacting and co-localizing with lamin A/C we also demonstrated that MLIP localizes to micro-domains in the nucleus with promyelocytic leukemia protein (PML) in close proximity to chromatin. MLIP's biological function still remains elusive. Eight week old hemizygous MLIP null mice develop enlarged hearts with a significant increase in heart to body weights (MLIP+/+ 5.62mg/g vs MLIP+/- 10.73mg/g, p<0.0001 n=7) with an overall 30% increase in the anterior-posterior ventricle length of MLIP hearts while maintaining a normal body weight (Figure). Echocardiographic analysis of MLIP+/- mice revealed that their hearts as having a significant (p3.93mm with a significant (p=0.011, n=12) reduction of left ventricular fractional shorting (LVFS) 31% when compare to littermate controls. Histological analysis of the hearts showed no overt phenotype other than an overall increase in the size of the MLIP+/- hearts. The cellular source for the increase in heart size and mass remains to be determined if it is the product of an increase in the number of cardiomyocytes due to aberrant hyperplasia or an increase in cardiomyocyte size through cellular hypertrophy. In conclusion, MLIP is a newly discovered lamin interacting protein that may serve as a transcriptional regulator that impact genes involved in heart development, growth and function and provides a new signaling paradigm.

Abstract 3: Il-10 Regulated Mir-375 Enhances Endothelial Progenitor Cell Mediated Myocardial Repair And Survival After Myocardial Infarction.

2014 ◽  
Vol 115 (suppl_1) ◽  
Author(s):  
Venkata N Garikipati ◽  
Prasanna Krishnamurthy ◽  
Suresh K Verma ◽  
Alexandra R Mackie ◽  
Erin E Vaughan ◽  
...  
Keyword(s):  
Capillary Density ◽  
Tube Formation ◽  
Lv Function ◽  
Myocardial Repair ◽  
Knock Down ◽  
Cardiac Functions ◽  
Dependent Protein ◽  
And Function ◽  
Deficient Mice

We hypothesized that IL-10 regulates miR-375 signaling in EPCs to enhance their survival and function in ischemic myocardium after MI. miR-375 knock down EPC were transplanted intramyocardially after induction of MI. Mice receiving EPC treated with miR-375 inhibitor showed increased number of GFP+EPCs retention that was associated with reduced EPC apoptosis in the myocardium. The engraftment of EPC into the vascular structures and the associated capillary density was significantly higher in miR-375-treated mice. The above findings further correlated with reduced infarct size, fibrosis and enhanced LV function (echocardiography) in miR-375 knock down EPC group as compared to scrambled EPC. Our in vitro studies revealed that the knockdown of miR-375 enhanced EPC proliferation, migration; tube formation ability and inhibited cell apoptosis, while the up-regulation of miR-375 with the mimic had the opposite effects. In addition, we found that miR-375 negatively regulates the expression of 3-phosphoinositide-dependent protein kinase 1 (PDK1) by directly targeting the 3'UTR of the PDK1 transcript. Interestingly, EPC isolated from IL-10-deficient mice has elevated basal levels of miR-375 and exhibited poor proliferation and tube formation ability where as miR-375 knock down in EPC isolated from IL-10 deficient mice attenuated these effects. Furthermore, transplantation of miR-375 knock down IL-10 deficient EPC after MI resulted in attenuated cardiac functions compared to scramble IL-10 deficient EPCs. Taken together, our studies suggest that IL-10 regulated miR-375 enhances EPC survival and function, associated with efficient myocardial repair via activation of PDK-1/AKT signaling cascades.

Abstract 37: Nrip Deficiency Leads to Dilated Cardiomyopathy

2015 ◽  
Vol 117 (suppl_1) ◽  
Author(s):  
Show-Li Chen
Keyword(s):  
Calcium Transient ◽  
Receptor Interaction ◽  
Cell Width ◽  
Interacting Protein ◽  
Cardiac Tissues ◽  
Calmodulin Binding ◽  
Calcineurin Phosphatase ◽  
And Function

Previously, we demonstrate a gene, nuclear receptor interaction protein (NRIP, also named DCAF6 or IQWD1) as a Ca2+- dependent calmodulin binding protein that can activate calcineurin phosphatase activity. Here, we found that α-actinin-2 (ACTN2), is one of NRIP-interacting proteins from the yeast two-hybrid system using NRIP as a prey. We further confirmed the direct bound between NRIP and ACTN2 using in vitro protein-protein interaction and in vivo co-immunoprecipitation assays. To further map the binding domain of each protein, the results showed the IQ domain of NRIP responsible for ACTN2 binding, and EF hand motif of ACTN2 responsible for NRIP bound. Due to ACTN2 is a biomarker of muscular Z-disc complex; we found the co-localization of NRIP and ACTN2 in cardiac tissues by immunofluorescence assays. Taken together, NRIP is a novel ACTN2-interacting protein. To investigate insights into in vivo function of NRIP, we generated conventional NRIP-null mice. The H&E staining results are shown in the hearts of NRIP KO mice are enlarged and dilated and the cell width of NRIP KO cardiomyocyte is increased. The EM of NRIP KO heart muscles reveal the reduction of I-band width and extension length of Z-disc in sarcomere structure; and the echocardiography shows the diminished fractional shortening in heart functions. Additionally, the calcium transient and sarcomere contraction length in cardiomyocytes of NRIP KO is weaker and shorter than wt; respectively. In conclusion, NRIP is a novel Z-disc protein and has function for maintenance of sarcomere integrity structure and function for calcium transient and muscle contraction.

scholarly journals Form and function in hillslope hydrology: characterization of subsurface flow based on response observations

2017 ◽  
Vol 21 (7) ◽  
pp. 3727-3748 ◽  
Author(s):  
Lisa Angermann ◽  
Conrad Jackisch ◽  
Niklas Allroggen ◽  
Matthias Sprenger ◽  
Erwin Zehe ◽  
...  
Keyword(s):  
Preferential Flow ◽  
Explanatory Power ◽  
Subsurface Flow ◽  
Time Lapse ◽  
Storm Event ◽  
Monitoring Methods ◽  
Catchment Scale ◽  
Flow Paths ◽  
Form And Function ◽  
And Function

Abstract. The phrase form and function was established in architecture and biology and refers to the idea that form and functionality are closely correlated, influence each other, and co-evolve. We suggest transferring this idea to hydrological systems to separate and analyze their two main characteristics: their form, which is equivalent to the spatial structure and static properties, and their function, equivalent to internal responses and hydrological behavior. While this approach is not particularly new to hydrological field research, we want to employ this concept to explicitly pursue the question of what information is most advantageous to understand a hydrological system. We applied this concept to subsurface flow within a hillslope, with a methodological focus on function: we conducted observations during a natural storm event and followed this with a hillslope-scale irrigation experiment. The results are used to infer hydrological processes of the monitored system. Based on these findings, the explanatory power and conclusiveness of the data are discussed. The measurements included basic hydrological monitoring methods, like piezometers, soil moisture, and discharge measurements. These were accompanied by isotope sampling and a novel application of 2-D time-lapse GPR (ground-penetrating radar). The main finding regarding the processes in the hillslope was that preferential flow paths were established quickly, despite unsaturated conditions. These flow paths also caused a detectable signal in the catchment response following a natural rainfall event, showing that these processes are relevant also at the catchment scale. Thus, we conclude that response observations (dynamics and patterns, i.e., indicators of function) were well suited to describing processes at the observational scale. Especially the use of 2-D time-lapse GPR measurements, providing detailed subsurface response patterns, as well as the combination of stream-centered and hillslope-centered approaches, allowed us to link processes and put them in a larger context. Transfer to other scales beyond observational scale and generalizations, however, rely on the knowledge of structures (form) and remain speculative. The complementary approach with a methodological focus on form (i.e., structure exploration) is presented and discussed in the companion paper by Jackisch et al.(2017).

scholarly journals Short-term post-mortality scavenging and longer term recovery after anoxia in the northern Adriatic Sea

2013 ◽  
Vol 10 (11) ◽  
pp. 7647-7659 ◽  
Author(s):  
M. Blasnig ◽  
B. Riedel ◽  
L. Schiemer ◽  
M. Zuschin ◽  
M. Stachowitsch
Keyword(s):  
Adriatic Sea ◽  
Time Lapse ◽  
Hermit Crabs ◽  
Small Scale ◽  
Short Term ◽  
Northern Adriatic Sea ◽  
Northern Adriatic ◽  
Hexaplex Trunculus ◽  
Distinct Sequence ◽  
And Function

Abstract. The northern Adriatic Sea is one of nearly 500 areas worldwide suffering widespread mortalities due to anoxia. The present study documents post-anoxia macrofauna dynamics after experimentally inducing small-scale anoxia in 24 m depth (2 plots, each 50 cm × 50 cm). Time-lapse camera deployments examined short-term scavenging of the moribund and dead organisms (multi-species clumps consisting of sponges and ascidians) over two 3-day periods (August 2009: 71.5 h, September 2009: 67.5 h). Longer term recovery (days to 2 yr) in the same two plots was examined with an independent photo series. Scavengers arrived quickly and in a distinct sequence: demersal (Gobius niger, Serranus hepatus) and benthopelagic fishes (Diplodus vulgaris, Pagellus erythrinus), followed by hermit crabs (Paguristes eremita, showing a clear day/night rhythm in presence) and gastropods (Hexaplex trunculus). This sequence is attributed to the relative speeds and densities of the organisms. The sessile fauna was largely removed or consumed within seven (August plot) and 13 (September plot) days after anoxia, confirming our first hypothesis that decaying organisms are quickly utilised. The scavengers remained in dense aggregations (e.g. up to 33 P. eremita individuals at one time) as long as dead organisms were available. No recovery of sessile macroepibenthos macroepibenthos occurred in the experimental plots one and two years after anoxia, undermining our second hypothesis that small denuded areas are more rapidly recolonised. This study underlines the sensitivity of this soft-bottom community and supports calls for reducing additional anthropogenic disturbances such as fishing practices that further impede recolonisation and threaten benthic community structure and function over the long term.

What makes vessels grow with exercise training?

2004 ◽  
Vol 97 (3) ◽  
pp. 1119-1128 ◽  
Author(s):  
Barry M. Prior ◽  
H. T. Yang ◽  
Ronald L. Terjung
Keyword(s):  
Vascular Remodeling ◽  
Tube Formation ◽  
Muscle Contractions ◽  
Endothelial Cell Proliferation ◽  
Mechanical Stimuli ◽  
Sprouting Angiogenesis ◽  
Intussusceptive Angiogenesis ◽  
Powerful Stimulus ◽  
Potent Growth ◽  
Tissue Reorganization

Exercise and muscle contractions create a powerful stimulus for structural remodeling of the vasculature. An increase in flow velocity through a vessel increases shear stress, a major stimulus for enlargement of conduit vessels. This leads to an endothelial-dependent, nitric oxide-dependent enlargement of the vessel. Increased flow within muscle, in the absence of contractions, leads to an enhanced capillarity by intussusceptive angiogenesis, a process of capillary splitting by intraluminal longitudinal divide. In contrast, sprouting angiogenesis requires extensive endothelial cell proliferation, with degradation of the extracellular matrix to permit migration and tube formation. This occurs during muscle adaptations to chronic contractions and/or muscle overload. The angiogenic growth factor VEGF appears to be an important element in angiogenesis. Recent advances in research have identified hemodynamic and mechanical stimuli that upregulate angiogenic processes, demonstrated a complexity of potent growth factors and interactions with their corresponding receptors, detected an interaction of cellular signaling events, and identified important tissue reorganization processes that must be coordinated to effect vascular remodeling. It is likely that much of this information is applicable to the vascular remodeling that occurs in response to exercise and/or muscle contractions.

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