Biology subjects

Heisenberg, C.-P.

Publications and source records attributed to Heisenberg, C.-P..

At least 19 recordsLinked to original sources

bioRxiv2025

Non-canonical cytokinesis driven by mechanical uncoupling via nematic flows and adhesion-based invagination

Cleavage - the series of rapid cell divisions that follow fertilization - marks the onset of metazoan development and represents a deeply conserved evolutionary process. Across animals, two principal modes exist: complete (holoblastic) and incomplete (meroblastic) cleavage. While holoblastic cleavage resembles conventional cytokinesis both in vitro and in vivo, the mechanisms underlying meroblastic cleavage have remained poorly understood. Using zebrafish embryos as a model, we show that meroblastic cleavage proceeds through a distinct two-step mechanism. The process begins with the assembly and contraction of a large, arc-shaped actomyosin cable. However, this contractile event alone is insufficient to complete division. A second phase, driven by cadherin-mediated membrane adhesion, is required to invaginate the furrow ridge. Strikingly, this transition depends on mechanical uncoupling of the contractile cable from the surrounding cortex. We demonstrate that such uncoupling arises from an active nematic instability, which both enhances contractility along the cable and generates actin depletion zones that relieve lateral connections. Together, these findings reveal that meroblastic cleavage is governed not by a single actomyosin-based event but by a sequential interplay between cytoskeletal contraction and cadherin-dependent adhesion, highlighting a mechanism fundamentally distinct from canonical cytokinesis.

cell biology↗

bioRxiv2025

The ancient metazoan cytoplasmic intermediate filament protein, Cilin, shapes cilia arrangement and tissue architecture

The emergence of animal multicellularity demanded novel cytoskeletal systems to support cellular architecture and tissue integrity. Cytoplasmic intermediate filaments (cIFs) are essential components of this scaffold, yet their evolutionary origins remain obscure. Here, we identify and characterize a lamin-derived, bona fide cIF protein in a ctenophore--a sister lineage to all other extant animals. We name this protein Cilin. Unlike nuclear lamins, Cilin lacks a nuclear localization signal and instead localizes predominantly to motile ciliary structures, including comb plates, the aboral organ, and sperm flagella, indicating a central role in ciliary architecture and function. Remarkably, Cilin is also present in non-ciliated cells, suggesting early functional diversification of cIFs in animal evolution. Phylogenetic and structural analyses position Cilin within an ancestral class of intermediate filament proteins, homologous to cnidarian and bilaterian nematocilins and lamin-tail-domain-containing (LMNTD) proteins. In humans, LMNTD proteins are enriched in ciliated epithelia and spermatids, pointing to a deeply conserved role in cilia-associated functions. These findings establish cilins as the earliest lamin-derived cytoplasmic intermediate filament proteins in metazoans, likely contributing to both ciliary function and the emergence of multicellular tissue organization.

cell biology↗

bioRxiv2025

Active and Probe-Free Intracellular Rheology via Phase-Sensitive Thermoviscous Flows

Determination of the rheological properties of cells is known to require active measurements, which largely depend on the internalisation of mechanical probes. Here we circumvent this problem via the introduction of Rheo-FLUCS: an active, yet probe-free approach that leverages light-induced flows to access the mechanics of complex systems. While Rheo-FLUCS is facilitated by thermoviscous expansion phenomena rather than external forces, we here show equivalence in its ability to measure canonical viscoelastic properties. Specifically, we demonstrate a phase-lag equivalence with probe-dependent active microrheology in a wide range of physically different, yet chemically identical materials. We exemplify the utility of Rheo-FLUCS in three distinctly different biological systems: compound-treated mouse fibroblasts (NIH-3T3), genetically modified human osteoblasts (U2OS) to elucidate the role of myosins in cytoplasmic mechanics, and early ascidian oocytes of Phallusia mammillata at fertilisation stage. This probe-free methodology represents a transformative advance in cellular rheology, enabling non-invasive and precise mechanical measurements across diverse biological systems without perturbing their natural state through probe internalisation.

biophysics↗

bioRxiv2025

Keratins coordinate tissue spreading by balancing spreading forces with tissue material properties

For tissues to spread, they must be deformable while maintaining their structural integrity. How these opposing requirements are balanced within spreading tissues is not yet well understood. Here, we show that keratin intermediate filaments function in epithelial spreading by adapting tissue mechanical resilience to the stresses arising in the tissue during the spreading process. By analysing the expansion of the enveloping cell layer (EVL) over the large yolk cell in early zebrafish embryos in vivo, we found that keratin network maturation in EVL cells is promoted by stresses building up within the spreading tissue. Through genetic interference and tissue rheology experiments, complemented by a vertex model with mechanochemical feedback, we demonstrate that stress-induced keratin network maturation in the EVL increases tissue viscosity, which is essential for preventing tissue rupture. Interestingly, keratins are also required in the yolk cell for mechanosensitive actomyosin network contraction and flow, the force-generating processes pulling the EVL. These dual mechanosensitive functions of keratins enable a balance between pulling force production in the yolk cell and the mechanical resilience of the EVL against stresses generated by these pulling forces, thereby ensuring uniform and robust tissue spreading.

developmental biology↗

bioRxiv2025

Geometry-driven asymmetric cell divisions pattern cell cycles and zygotic genome activation in the zebrafish embryo

Early embryo geometry is one of the most invariant species-specific traits, yet its role in ensuring developmental reproducibility and robustness remains underexplored. Here, we show that in zebrafish, the geometry of the fertilized egg - specifically its curvature and volume - serves as a critical initial condition triggering a cascade of events that exert a lasting influence on development. Specifically, it guides asymmetric cell divisions in the blastoderm, generating radial gradients of cell volume and nucleocytoplasmic ratio. These gradients generate mitotic phase waves, with individual cell cycle periods determined largely cell-autonomously by the nucleocytoplasmic ratio. Modeling and perturbation experiments demonstrate that reducing cell autonomy reshapes these waves, emphasizing the instructive role of geometry-derived volume patterns in setting the intrinsic period of the cell cycle oscillator. Remarkably, in addition to organizing cell cycles, early embryo geometry also spatially patterns zygotic genome activation (ZGA) at the midblastula transition, a key step in establishing embryonic autonomy. Disrupting geometry alters the ZGA pattern and causes ectopic germ layer specification, underscoring its developmental significance. Together, our findings reveal a novel symmetry-breaking function of early embryo geometry in coordinating cell cycle and transcriptional patterning, establishing a blueprint for robust embryogenesis.

developmental biology↗

bioRxiv2024

hoxb genes determine the timing of cell ingression by regulating cell surface fluctuations during zebrafish gastrulation

During embryonic development, cell behaviors need to be tightly regulated in time and space. Yet, how the temporal and spatial regulations of cell behaviors are interconnected during embryonic development remains elusive. To address this, we turned to zebrafish gastrulation, the process where dynamic cell behaviors generate the three principal germ layers of the early embryo. Here, we show that hoxb cluster genes are expressed in a temporally collinear manner at the blastoderm margin of the embryo to regulate the timing of mesoderm and endoderm (mesendoderm) cell ingression. Under- or over-expression of hoxb genes perturb the timing of mesendoderm cell ingression and, consequently, the positioning of these cells along the forming anterior-posterior body axis. Finally, we found that hoxb genes control the timing of mesendoderm ingression by regulating cell surface fluctuations. Thus, hoxb genes interconnect the temporal and spatial pattern of cell behavior in the early embryo by controlling cell surface fluctuations. HighlightsO_LIhoxb gene expression shows temporal collinearity at the blastoderm margin during zebrafish gastrulation. C_LIO_LITemporal collinear expression of hoxb genes at the blastoderm margin regulates the timing of cell ingression and delineates spatial collinearity after gastrulation. C_LIO_LIhoxb genes regulate cell surface fluctuations and bleb formation at the blastoderm margin during cell ingression C_LI Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=149 SRC="FIGDIR/small/579056v1_ufig1.gif" ALT="Figure 1"> View larger version (37K): org.highwire.dtl.DTLVardef@3a8517org.highwire.dtl.DTLVardef@1f7bda3org.highwire.dtl.DTLVardef@1c325a6org.highwire.dtl.DTLVardef@dce24d_HPS_FORMAT_FIGEXP M_FIG C_FIG hoxb genes are expressed at the blastoderm margin in a temporally collinear manner and determine the timing of mesendoderm ingression by triggering dynamic cell surface fluctuations and cell blebbing. This results in a spatial collinearity of mesendoderm cell positioning along the anterior-posterior extent of the forming body axis with early ingressing cells being positioned more anteriorly than late ingressing cells.

developmental biology↗

bioRxiv2023

BMP-dependent patterning of ectoderm tissue material properties modulates lateral mesendoderm cell migration during early zebrafish gastrulation

Cell migration is a fundamental process during embryonic development. Most studies on cell migration in vivo have focussed on the migration of cells using the extracellular matrix (ECM) as their substrate for migration. In contrast, much less is known about how cells migrate on other cells, as found in early embryos when the ECM has not yet formed. Here, we show that lateral mesendoderm (LME) cells in the early zebrafish gastrula use the overlying ectoderm as their substrate for migration. We show that the lateral ectoderm is permissive for the animal pole-directed migration of LME cells, while the ectoderm at the animal pole of the gastrula halts animal pole-directed LME migration. These differences in the permissive properties of the ectoderm for LME migration are due to the lateral ectoderm being more cohesive and viscous than the animal ectoderm. Consistently, tuning cell contractility in lateral and animal ectoderm via modulation of the actomyosin cytoskeleton is sufficient to change their permissiveness for LME migration. Finally, we found that BMP signalling is critical for reducing animal ectoderm cohesion and, thus, its capacity to halt LME migration towards the animal pole. Collectively, these findings identify a critical role of ectoderm tissue cohesion and viscosity in guiding LME migration during zebrafish gastrulation.

developmental biology↗

bioRxiv2023

Robust axis elongation by Nodal-dependent restriction of BMP signaling

Embryogenesis is brought about by the coordinated activities of different signaling pathways controlling cell fate specification and morphogenesis. In vertebrate gastrulation, both Nodal and BMP signaling play key roles in germ layer specification and morphogenesis, yet their interplay to coordinate embryo patterning with morphogenesis is still insufficiently understood. Here, we took a reductionist approach using zebrafish embryonic explants to study the coordination of Nodal and BMP signaling for embryo patterning and morphogenesis. We show that Nodal signaling not only triggers explant elongation by inducing mesendodermal progenitors but also by suppressing BMP signaling activity at the site of mesendoderm induction. Ectopic BMP signaling in the mesendoderm blocks cell alignment and oriented mesendoderm intercalations, key processes to drive explant elongation. Translating these ex vivo observations to the intact zebrafish embryo showed that, similar to explants, Nodal signaling renders the dorsal domain less sensitive towards BMP signaling to allow effective cell intercalations and thus robust embryonic axis elongation. These findings suggest a dual function of Nodal signaling in embryonic axis elongation by both inducing mesendoderm and maintaining low levels of BMP signaling activity in the dorsal portion of the mesendoderm.

developmental biology↗

bioRxiv2023

Adhesion-induced cortical flows pattern E-cadherin-mediated cell contacts

Metazoan development relies on the formation and remodeling of cell-cell contacts. Dynamic reorganization of adhesion receptors and the actomyosin cell cortex in space and time play a central role in cell-cell contact formation and maturation. Yet, how this process is mechanistically achieved remains unclear. Here, by building a biomimetic assay composed of progenitor cells adhering to supported lipid bilayers functionalized with E-cadherin ectodomains, we show that cortical Actin flows, driven by the depletion of Myosin-2 at the cell contact center, mediate the dynamic reorganization of adhesion receptors and cell cortex at the contact. E-cadherin-dependent downregulation of the small GTPase RhoA at the forming contact leads to both a depletion of Myosin-2 and a decrease of F-actin at the contact center. This depletion of Myosin-2 causes centrifugal F-actin flows, leading to further accumulation of F-actin at the contact rim and the progressive redistribution of E-cadherin from the contact center to the rim. Eventually, this combination of actomyosin downregulation and flows at the contact determine the characteristic molecular organization, with E-cadherin and F-actin accumulating at the contact rim, where they are needed to mechanically link the contractile cortices of the adhering cells.

cell biology↗

bioRxiv2023

Mechanical forces in extendable tissue matrix orient cell divisions via microtubule stabilization in Arabidopsis

Plant morphogenesis relies exclusively on oriented cell expansion and division. Nonetheless, the mechanism(s) determining division plane orientation remain elusive. Here we studied tissue healing after laser-assisted wounding in roots and uncovered how mechanical forces of cell expansion stabilize and reorient microtubule cytoskeleton for orientation of cell division. We revealed that root tissue functions as interconnected cell matrix with a radial gradient of tissue extendibility causing a predictable tissue deformation after wounding. This causes instant redirection of expansion in the surrounding cells and reorientation of microtubule arrays ultimately predicting cell division orientation. Microtubules are destabilized under low tension, whereas stretching of cells, either through wounding or external aspiration immediately induce their polymerisation. The higher microtubule abundance in the stretched cell parts leads to reorientation microtubule arrays and ultimately cell division planes. This provides a long-sought mechanism for flexible re-arrangement of cell divisions by mechanical forces for tissue reconstruction and plant architecture.

cell biology↗

bioRxiv2022

Zinc-based Ultrasensitive Microscopic Barrier Assay (ZnUMBA): a live-imaging method for detecting epithelial barrier breaches with spatiotemporal precision

Epithelial barrier function is commonly analyzed using transepithelial electrical resistance (TER), which measures the ion flux across epithelia, or by adding traceable macromolecules to one side of the epithelium and monitoring their passage to the other side. While these methods effectively measure changes to global barrier function, they are not sensitive enough to detect local or transient disruptions in the barrier, and they do not reveal the location of barrier breaches within the context of cell or tissue morphology. Therefore, we developed a method that we named Zinc-based Ultrasensitive Microscopic Barrier Assay (ZnUMBA), which overcomes these limitations, allowing for detection of local tight junction (TJ) leaks with high spatial and temporal resolution (Stephenson et al., 2019; Varadarajan et al., 2021). Here, we present expanded applications for ZnUMBA. First, we show that ZnUMBA can be used in Xenopus embryos to measure the dynamics of barrier restoration and actin dynamics following laser injury of the junction. We also demonstrate that ZnUMBA can be effectively utilized in developing zebrafish embryos as well as cultured monolayers of Madin-Darby Canine Kidney II (MDCK II) epithelial cells. ZnUMBA is a powerful and flexible method that, with some optimization, can be applied to multiple systems to measure dynamic changes in barrier function with spatiotemporal precision.

cell biology↗

bioRxiv2022

Yolk granule fusion and microtubule aster formation regulate cortical granule translocation and exocytosis in zebrafish oocytes

Dynamic reorganization of the cytoplasm is key to many core cellular processes, such as cell division, cell migration and cell polarization. Cytoskeletal rearrangements are thought to constitute the main drivers of cytoplasmic flows and reorganization. In contrast, remarkably little is known about how dynamic changes in size and shape of cell organelles affect large-scale cytoplasmic organization. Here, we show that within the maturing zebrafish oocyte, the surface localization of exocytosis-competent cortical granules upon germinal vesicle breakdown is achieved by the combined activities of yolk granule fusion and microtubule aster formation and translocation. We find that cortical granules are moved towards the oocyte surface through radially-outward cytoplasmic flows induced by yolk granules fusing within the oocyte center in response to GV breakdown. We further show that vesicles decorated with the small Rab GTPase Rab11, a master regulator of vesicular trafficking and exocytosis, accumulate together with cortical granules at the oocyte surface. This accumulation is achieved by Rab11-positive vesicles being transported by acentrosomal microtubule asters, the formation of which is induced by the release of CyclinB/Cdk1 upon GV breakdown, and which display a net movement towards the oocyte surface by preferentially binding to the oocyte actin cortex. We finally demonstrate that the decoration of cortical granules by Rab11 at the oocyte surface is needed for cortical granule release and subsequent chorion elevation, a process central in oocyte activation. Collectively, these findings unravel a yet unrecognized role of organelle fusion, functioning together with cytoskeletal rearrangements, in determining cytoplasmic organization during oocyte maturation.

developmental biology↗

bioRxiv2022

Morphogen gradient orchestrates pattern-preserving tissue morphogenesis via motility-driven (un)jamming

Embryo development requires both biochemical signalling generating patterns of cell fates and active mechanical forces driving tissue shape changes. Yet, how these fundamental processes are coordinated in space and time, and, especially, how tissue patterning is preserved despite the complex cellular flows occurring during morphogenesis, remains poorly understood. Here, we show that a Nodal/TGF-{beta} morphogen gradient orchestrates pattern-preserving mesendoderm internalization movements during zebrafish gastrulation by triggering a motility-driven (un)jamming transition. We find that graded Nodal signalling, in addition to its highly conserved role in mesendoderm patterning, mechanically subdivides the tissue into a small fraction of highly protrusive leader cells able to locally unjam and thus autonomously internalize, and less protrusive followers, which remain jammed and need to be pulled inwards by the leaders. Using minimal particle-based simulations and experimental perturbations, we further show that this binary mechanical switch, when combined with Nodal-dependent preferential adhesion coupling leaders to followers, is critical for triggering collective and orderly mesendoderm internalization, thus preserving tissue patterning. This provides a simple, yet quantitative, theoretical framework for how a morphogen-encoded (un)jamming transition can bidirectionally couple tissue mechanics with patterning during complex three-dimensional morphogenesis.

developmental biology↗

bioRxiv2021

Admp regulates tail bending by controlling the intercalation of the ventral epidermis through myosin phosphorylation.

The transient but pronounced ventral tail bending is found in many chordate embryos and constitutes an interesting model of how tissue interactions control embryo shape (Lu et al., 2020). Here, we identify one key upstream regulator of ventral tail bending in the ascidian Ciona embryo. We show that during early tailbud stage, ventral epidermal cells exhibit a boat-shaped morphology (boat cell) with a narrow apical surface where phosphorylated myosin (pMLC) accumulated. We further show that interfering with the function of the BMP ligand Admp leads to pMLC localizing to the basal instead of the apical side of ventral epidermal cells and a reduced number of boat cells. Finally, we show that cutting ventral epidermal midline cells at their apex using a ultraviolet laser relaxes ventral tail bending. Based on these results, we propose a novel function for Admp in localizing pMLC to the apical side of ventral epidermal cells, which causes the tail to bend ventrally by resisting antero-posterior notochord extension at the ventral side of the tail. Summary StatementAdmp is an upstream regulator of tail bending in the chordate Ciona tailbud embryo, determining tissue polarity of the ventral midline epidermis by localizing phosphorylated myosin.

developmental biology↗

bioRxiv2021

Multi-tier mechanics control stromal adaptations in swelling lymph nodes

Lymph nodes (LNs) comprise two main structural elements: Fibroblastic reticular cells (FRCs) that form dedicated niches for immune cell interaction and capsular fibroblasts that build a shell around the organ. While LNs are fairly stable in size during homeostatic conditions, immunological challenge causes more than 10-fold increase in size within only a few days. How a solid organ can accommodate such extreme volumetric changes is poorly understood. Here, we characterize the biomechanics of LN swelling on the cellular and organ scale. We identify lymphocyte trapping by influx and proliferation as drivers of an outward pressure force, causing FRCs and their associated conduits to stretch. After an initial phase of relaxation, FRCs sense the resulting strain via cell matrix adhesions, which coordinates local growth and remodeling of the stromal network. While the expanded FRC network adopts its typical configuration, a massive fibrotic reaction of the organ capsule sets in and counters further organ expansion. Thus, different fibroblast populations mechanically control LN swelling in a multi-tier fashion.

immunology↗

bioRxiv2021

Combined effect of cell geometry and polarity domains determines the orientation of unequal division

Cell division orientation is thought to result from a competition between cell geometry and polarity domains controlling the position of the mitotic spindle during mitosis. Depending on the level of cell shape anisotropy or the strength of the polarity domain, one dominates the other and determines the orientation of the spindle. Whether and how such competition is also at work to determine unequal cell division (UCD), producing daughter cells of different size, remains unclear. Here, we show that cell geometry and polarity domains cooperate, rather than compete, in positioning the cleavage plane during UCDs in early ascidian embryos. We found that the UCDs and their orientation at the ascidian third cleavage rely on the spindle tilting in an anisotropic cell shape, and cortical polarity domains exerting different effects on spindle astral microtubules. By systematically varying mitotic cell shape, we could modulate the effect of attractive and repulsive polarity domains and consequently generate predicted daughter cell size asymmetries and position. We therefore propose that the spindle position during UCD is set by the combined activities of cell geometry and polarity domains, where cell geometry modulates the effect of cortical polarity domain(s). Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=200 SRC="FIGDIR/small/442692v1_ufig1.gif" ALT="Figure 1"> View larger version (75K): org.highwire.dtl.DTLVardef@4b0e87org.highwire.dtl.DTLVardef@1a6fdbdorg.highwire.dtl.DTLVardef@5ca0ddorg.highwire.dtl.DTLVardef@1450e61_HPS_FORMAT_FIGEXP M_FIG C_FIG HighlightO_LISpindle tilting in anisotropic cell shape induces unequal cell division C_LIO_LICortical polarity domain can exert attractive or repulsive effect on spindle C_LIO_LICell geometry and polarity domain cooperate to position the spindle C_LIO_LICell geometry modulates the effect of polarity domain C_LI

cell biology↗

bioRxiv2021

Apical contacts stemming from incomplete delamination guide progenitor cell allocation through a dragging mechanism

The developmental strategies used by progenitor cells to endure a safe journey from their induction place towards the site of terminal differentiation are still poorly understood. Here we uncovered a progenitor cell allocation mechanism that stems from an incomplete process of epithelial delamination that allows progenitors to coordinate their movement with adjacent extra-embryonic tissues. Progenitors of the zebrafish laterality organ originate from the surface epithelial enveloping layer by an apical constriction process of cell delamination. During this process, progenitors retain long-term apical contacts that enable the epithelial layer to pull a subset of progenitors along their way towards the vegetal pole. The remaining delaminated progenitors follow apically-attached progenitors movement by a co-attraction mechanism, avoiding sequestration by the adjacent endoderm, ensuring their fate and collective allocation at the differentiation site. Thus, we reveal that incomplete delamination serves as a cellular platform for coordinated tissue movements during development. Impact StatementIncomplete delamination serves as a cellular platform for coordinated tissue movements during development, guiding newly formed progenitor cell groups to the differentiation site.

developmental biology↗

bioRxiv2020

Satb2 acts as a gatekeeper for major developmental transitions during early vertebrate embryogenesis

Zygotic genome activation (ZGA) initiates regionalized transcription responsible for the acquisition of distinct cellular identities. ZGA is dependent upon dynamic chromatin architecture sculpted by conserved DNA-binding proteins. However, whether the tissue-specific transcription is mechanistically linked with the onset of ZGA is unknown. Here, we have addressed the involvement of chromatin organizer SATB2 in orchestrating these processes during vertebrate embryogenesis. Integrative analysis of transcriptome, genome-wide occupancy and chromatin accessibility revealed contrasting molecular functions of maternal and zygotic pools of Satb2. Maternal Satb2 represses zygotic genes by influencing the interplay between the pluripotency factors. By contrast, zygotic Satb2 activates transcription of the same group of genes during neural crest development and organogenesis. Comparative analysis of maternal versus zygotic function of Satb2 underscores how these antithetical activities are temporally coordinated and functionally implemented. We discuss the evolutionary implications of the biphasic and bimodal regulation of landmark developmental transitions by a single determinant.

developmental biology↗