scholarly journals Twisting of the zebrafish heart tube during cardiac looping is a tbx5-dependent and tissue-intrinsic process

eLife ◽  
2021 ◽  
Vol 10 ◽  
Author(s):  
Federico Tessadori ◽  
Erika Tsingos ◽  
Enrico Sandro Colizzi ◽  
Fabian Kruse ◽  
Susanne C van den Brink ◽  
...  
Keyword(s):  
Ex Vivo ◽  
Loss Of Function ◽  
Heart Tube ◽  
Atrioventricular Canal ◽  
Internal Organs ◽  
Cardiac Looping ◽  
Tissue Patterning ◽  
Ex Vivo Culture ◽  
Left Right Asymmetry ◽  
Organ Laterality

Organ laterality refers to the left-right asymmetry in disposition and conformation of internal organs and is established during embryogenesis. The heart is the first organ to display visible left-right asymmetries through its left-sided positioning and rightward looping. Here, we present a new zebrafish loss-of-function allele for tbx5a, which displays defective rightward cardiac looping morphogenesis. By mapping individual cardiomyocyte behavior during cardiac looping, we establish that ventricular and atrial cardiomyocytes rearrange in distinct directions. As a consequence, the cardiac chambers twist around the atrioventricular canal resulting in torsion of the heart tube, which is compromised in tbx5a mutants. Pharmacological treatment and ex vivo culture establishes that the cardiac twisting depends on intrinsic mechanisms and is independent from cardiac growth. Furthermore, genetic experiments indicate that looping requires proper tissue patterning. We conclude that cardiac looping involves twisting of the chambers around the atrioventricular canal, which requires correct tissue patterning by Tbx5a.

scholarly journals Twisting of the heart tube during cardiac looping is a tbx5-dependent and tissue-intrinsic process

2020 ◽  
Author(s):  
Federico Tessadori ◽  
Fabian Kruse ◽  
Susanne C. van den Brink ◽  
Malou van den Boogaard ◽  
Vincent M. Christoffels ◽  
...  
Keyword(s):  
Ex Vivo ◽  
Tissue Growth ◽  
Confocal Imaging ◽  
Loss Of Function ◽  
Heart Tube ◽  
Atrioventricular Canal ◽  
Cardiac Looping ◽  
Tissue Patterning ◽  
Heart Looping ◽  
Organ Laterality

AbstractOrgan laterality refers to the Left-Right (LR) asymmetry in disposition and conformation of internal organs, established in the developing embryo. The heart is the first organ to display visible LR asymmetries as it is positioned to the left side of the midline and undergoes rightward looping morphogenesis. Cardiac looping morphogenesis is tightly controlled by a combination of heart-intrinsic and -extrinsic mechanisms. As the mechanisms that drive cardiac looping are not well understood, we performed a forward genetic screen for zebrafish mutants with defective heart looping. We describe a new loss-of-function allele for tbx5a, which displays normal leftward positioning but defective rightward looping morphogenesis. By using live two-photon confocal imaging to map cardiomyocyte behavior during cardiac looping at a single-cell level we establish that during looping, ventricular and atrial cardiomyocytes rearrange in opposite directions towards the outer curvatures of the chambers. As a consequence, the cardiac chambers twist around the atrioventricular canal resulting in torsion of the heart tube, which is compromised in tbx5a mutants. Manipulations of cardiac looping by chemical treatment and ex vivo culture establishes that the twisting of the heart tube depends on intrinsic mechanisms and is independent from tissue growth by cell addition. Furthermore, the cardiac looping defect in tbx5a mutants is rescued in tbx5a/tbx2b double mutants, indicating that it requires proper tissue patterning. Together, our results establish that cardiac looping in zebrafish involves twisting of the chambers around the AV canal, which requires correct tissue patterning by Tbx5a.

scholarly journals Genetic Steps to Organ Laterality in Zebrafish

2001 ◽  
Vol 2 (2) ◽  
pp. 60-68 ◽  
Author(s):  
Jau-Nian Chen ◽  
Frauke van Bebber ◽  
Allan M. Goldstein ◽  
Fabrizio C. Serluca ◽  
Donald Jackson ◽  
...  
Keyword(s):  
Genetic Screen ◽  
Visceral Organ ◽  
Heart Tube ◽  
Internal Organs ◽  
Body Form ◽  
Recessive Mutations ◽  
Class 1 ◽  
Liver And Pancreas ◽  
Left Right Asymmetry ◽  
Organ Laterality

All internal organs are asymmetric along the left–right axis. Here we report a genetic screen to discover mutations which perturb organ laterality. Our particular focus is upon whether, and how, organs are linked to each other as they achieve their laterally asymmetric positions. We generated mutations by ENU mutagenesis and examined F3 progeny using a cocktail of probes that reveal early primordia of heart, gut, liver and pancreas. From the 750 genomes examined, we isolated seven recessive mutations which affect the earliest left–right positioning of one or all of the organs. None of these mutations caused discernable defects elsewhere in the embryo at the stages examined. This is in contrast to those mutations we reported previously (Chenet al., 1997) which, along with left–right abnormalities, cause marked perturbation in gastrulation, body form or midline structures. We find that the mutations can be classified on the basis of whether they perturb relationships among organ laterality. In Class 1 mutations, none of the organs manifest any left–right asymmetry. The heart does not jog to the left and normally leftpredominantBMP4in the early heart tube remains symmetric. The gut tends to remain midline. There frequently is a remarkable bilateral duplication of liver and pancreas. Embryos with Class 2 mutations have organotypic asymmetry but, in any given embryo, organ positions can be normal, reversed or randomized. Class 3 reveals a hitherto unsuspected gene that selectively affects laterality of heart. We find that visceral organ positions are predicted by the direction of the preceding cardiac jog. We interpret this as suggesting that normally there is linkage between cardiac and visceral organ laterality. Class 1 mutations, we suggest, effectively remove the global laterality signals, with the consequence that organ positions are effectively symmetrical. Embryos with Class 2 mutations do manifest linkage among organs, but it may be reversed, suggesting that the global signals may be present but incorrectly orientated in some of the embryos. That laterality decisions of organs may be independently perturbed, as in the Class 3 mutation, indicates that there are distinctive pathways for reception and organotypic interpretation of the global signals.

scholarly journals Asymmetric Hapln1a drives regionalised cardiac ECM expansion and promotes heart morphogenesis during zebrafish development

2019 ◽  
Author(s):  
Christopher J Derrick ◽  
Juliana Sánchez-Posada ◽  
Farah Hussein ◽  
Federico Tessadori ◽  
Eric JG Pollitt ◽  
...  
Keyword(s):  
Cross Linking ◽  
Heart Tube ◽  
Cardiac Morphogenesis ◽  
Heart Morphogenesis ◽  
Cardiac Looping ◽  
Intrinsic Signals ◽  
Link Protein ◽  
Specific Expansion ◽  
Left Right Asymmetry

AbstractThe mature vertebrate heart develops from a simple linear cardiac tube during early development through a series of highly asymmetric morphogenetic processes including cardiac looping and chamber ballooning. While the directionality of heart morphogenesis is partly controlled by embryonic laterality signals, previous studies have suggested that these extrinsic laterality cues interact with tissue-intrinsic signals in the heart to ensure robust asymmetric cardiac morphogenesis. Using live in vivo imaging of zebrafish embryos we describe a left-sided, chamber-specific expansion of the extracellular matrix (ECM) between the myocardium and endocardium at early stages of heart morphogenesis. We use Tomo-seq, a spatial transcriptomic approach, to identify transient and regionalised expression of hyaluronan and proteoglycan link protein 1a (hapln1a), encoding an ECM cross-linking protein, in the heart tube prior to cardiac looping overlapping with regionalised ECM expansion. Loss- and gain-of-function experiments demonstrate that regionalised Hapln1a promotes heart morphogenesis through regional modulation of ECM thickness in the heart tube. Finally, we show that while induction of asymmetric hapln1a expression is independent of embryonic left-right asymmetry, these laterality cues are required to orient the hapln1a-expressing cells asymmetrically along the left-right axis of the heart tube.Together, we propose a model whereby laterality cues position hapln1a expression on the left of the heart tube, and this asymmetric Hapln1a deposition drives ECM asymmetry and subsequently promotes robust asymmetric cardiac morphogenesis.

Failure of ventral closure and axial rotation in embryos lacking the proprotein convertase Furin

Development ◽  
1998 ◽  
Vol 125 (24) ◽  
pp. 4863-4876 ◽  
Author(s):  
A.J. Roebroek ◽  
L. Umans ◽  
I.G. Pauli ◽  
E.J. Robertson ◽  
F. van Leuven ◽  
...  
Keyword(s):  
Axial Rotation ◽  
Marker Genes ◽  
Loss Of Function ◽  
Heart Tube ◽  
Lateral Plate ◽  
Form Analysis ◽  
Left Right Asymmetry ◽  
Gut Endoderm ◽  
Endothelial Precursors

We have examined the role of Furin in postimplantation-stage mouse embryos by analyzing both the expression pattern of fur mRNA and the developmental consequences of a loss-of-function mutation at the fur locus. At early stages (day 7.5), fur mRNA is abundant in extraembryonic endoderm and mesoderm, anterior visceral endoderm, and in precardiac mesoderm. 1 day later fur is expressed throughout the heart tube and in the lateral plate mesoderm, notochordal plate and definitive gut endoderm. Embryos lacking Furin die between days 10.5 and 11.5, presumably due to hemodynamic insufficiency associated with severe ventral closure defects and the failure of the heart tube to fuse and undergo looping morphogenesis. Morphogenesis of the yolk sac vasculature is also abnormal, although blood islands and endothelial precursors form. Analysis of cardiac and endodermal marker genes shows that while both myocardial precursors and definitive endoderm cells are specified, their numbers and migratory properties are compromised. Notably, mutant embryos fail to undergo axial rotation, even though Nodal and eHand, two molecular markers of left-right asymmetry, are appropriately expressed. Overall, the present data identify Furin as an important activator of signals responsible for ventral closure and embryonic turning.

Hyperactivation of RAP1 and JAK/STAT Signaling Pathways Contributes to Fibrosis during the Formation of Nasal Capsular Contraction

2021 ◽  
pp. 1-12
Author(s):  
Meng Wu ◽  
Ming Li ◽  
Hong-Ju Xie ◽  
Hong-Wei Liu
Keyword(s):  
Signaling Pathways ◽  
Type I Collagen ◽  
Ex Vivo ◽  
Capsular Contracture ◽  
Silicone Implant ◽  
Type I ◽  
Loss Of Function ◽  
Sequencing Data ◽  
Functional Changes ◽  
Stat Signaling

Silicone implant-based augmentation rhinoplasty or mammoplasty induces capsular contracture, which has been acknowledged as a process that develops an abnormal fibrotic capsule associated with the immune response to allogeneic materials. However, the signaling pathways leading to the nasal fibrosis remain poorly investigated. We aimed to explore the molecular mechanism underlying the pathogenesis of nasal capsular contracture, with a specific research interest in the signaling pathways involved in fibrotic development at the advanced stage of contracture. By examining our recently obtained RNA sequencing data and global gene expression profiling between grade II and grade IV nasal capsular tissues, we found that both the RAP1 and JAK/STAT signaling pathways were hyperactive in the contracted capsules. This was verified on quantitative real-time PCR which demonstrated upregulation of most of the representative component signatures in these pathways. Loss-of-function assays through siRNA-mediated Rap1 silencing and/or small molecule-directed inhibition of JAK/STAT pathway in ex vivo primary nasal fibroblasts caused a series of dramatic behavioral and functional changes, including decreased cell viability, increased apoptosis, reduced secretion of proinflammatory cytokines, and synthesis of type I collagen, compared to control cells, and indicating the essential role of the RAP1 and JAK/STAT signaling pathways in nasal capsular fibrosis. Our results sheds light on targeting downstream signaling pathways for the prevention and therapy of silicone implant-induced nasal capsular contracture.

Phagocytosis of Liposome Particles by Rat Splenic Immature Monocytes Makes Them Transiently and Highly Immunosuppressive In Ex Vivo Culture Conditions

2011 ◽  
Vol 337 (1) ◽  
pp. 42-49 ◽  
Author(s):  
Daisuke Takahashi ◽  
Hiroshi Azuma ◽  
Hiromi Sakai ◽  
Keitaro Sou ◽  
Daiko Wakita ◽  
...  
Keyword(s):  
Ex Vivo ◽  
Culture Conditions ◽  
Ex Vivo Culture

scholarly journals Mimicking of Blood Flow Results in a Distinct Functional Phenotype in Human Non-Adherent Classical Monocytes

Biology ◽  
2021 ◽  
Vol 10 (8) ◽  
pp. 748
Author(s):  
Elisa Wirthgen ◽  
Melanie Hornschuh ◽  
Ida Maria Wrobel ◽  
Christian Manteuffel ◽  
Jan Däbritz
Keyword(s):  
Blood Flow ◽  
Shear Flow ◽  
Ex Vivo ◽  
Culture Conditions ◽  
Inflammatory Signaling ◽  
Gm Csf ◽  
Beneficial Effects ◽  
Migratory Capacity ◽  
Ex Vivo Culture ◽  
Static Conditions

Ex vivo culture conditions during the manufacturing process impact the therapeutic effect of cell-based products. Mimicking blood flow during ex vivo culture of monocytes has beneficial effects by preserving their migratory ability. However, the effects of shear flow on the inflammatory response have not been studied so far. Hence, the present study investigates the effects of shear flow on both blood-derived naïve and activated monocytes. The activation of monocytes was experimentally induced by granulocyte-macrophage colony-stimulating factor (GM-CSF), which acts as a pro-survival and growth factor on monocytes with a potential role in inflammation. Monocytes were cultured under dynamic (=shear flow) or static conditions while preventing monocytes' adherence by using cell-repellent surfaces to avoid adhesion-induced differentiation. After cultivation (40 h), cell size, viability, and cytokine secretion were evaluated, and the cells were further applied to functional tests on their migratory capacity, adherence, and metabolic activity. Our results demonstrate that the application of shear flow resulted in a decreased pro-inflammatory signaling concurrent with increased secretion of the anti-inflammatory cytokine IL-10 and increased migratory capacity. These features may improve the efficacy of monocyte-based therapeutic products as both the unwanted inflammatory signaling in blood circulation and the loss of migratory ability will be prevented.

scholarly journals Stem cell integrins: Implications for ex-vivo culture and cellular therapies

2011 ◽  
Vol 6 (1) ◽  
pp. 1-12 ◽  
Author(s):  
Andrew B.J. Prowse ◽  
Fenny Chong ◽  
Peter P. Gray ◽  
Trent P. Munro
Keyword(s):  
Stem Cell ◽  
Ex Vivo ◽  
Cellular Therapies ◽  
Ex Vivo Culture

scholarly journals Ex vivo culture of Fancc-/- stem/progenitor cells predisposes cells to undergo apoptosis, and surviving stem/progenitor cells display cytogenetic abnormalities and an increased risk of malignancy

Blood ◽  
2005 ◽  
Vol 105 (9) ◽  
pp. 3465-3471 ◽  
Author(s):  
Xiaxin Li ◽  
Michelle M. Le Beau ◽  
Samantha Ciccone ◽  
Feng-Chun Yang ◽  
Brian Freie ◽  
...  
Keyword(s):  
Progenitor Cells ◽  
Genome Stability ◽  
Bone Marrow Cells ◽  
Ex Vivo ◽  
Acquired Resistance ◽  
Intrinsic Defects ◽  
Cytogenetic Abnormalities ◽  
Increased Risk ◽  
Ex Vivo Culture ◽  
The Impact

AbstractCurrent strategies for genetic therapy using Moloney retroviruses require ex vivo manipulation of hematopoietic cells to facilitate stable integration of the transgene. While many studies have evaluated the impact of ex vivo culture on normal murine and human stem/progenitor cells, the cellular consequences of ex vivo manipulation of stem cells with intrinsic defects in genome stability are incompletely understood. Here we show that ex vivo culture of Fancc-/- bone marrow cells results in a time-dependent increase in apoptosis of primitive Fancc-/- progenitor cells in conditions that promote the proliferation of wild-type stem/progenitor cells. Further, recipients reconstituted with the surviving Fancc-/- cells have a high incidence of cytogenetic abnormalities and myeloid malignancies that are associated with an acquired resistance to tumor necrosis factor α (TNF-α). Collectively, these data indicate that the intrinsic defects in the genomic stability of Fancc-/- stem/progenitor cells provide a selective pressure for cells that are resistant to apoptosis and have a propensity for the evolution to clonal hematopoiesis and malignancy. These studies could have implications for the design of genetic therapies for treatment of Fanconi anemia and potentially other genetic diseases with intrinsic defects in genome stability.

Sign in / Sign up

Export Citation Format

Share Document