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Lenz, M. O.

Publications and source records attributed to Lenz, M. O..

12 recordsLinked to original sources

Live confocal imagining of cellular touch responses upon local quantifiable mechanical stimulation in plant cells

Plant cells grow and differentiate in an ever-changing environment characterized by transient signals and stimuli. The ability of plant cells to perceive, integrate, and dynamically respond to these stimuli underpins a plant adaptation and survival. Among the plethora of complex stimuli plant cells are exposed to, several stimuli have a mechanical component, for example, wind, touch, contact with insects, penetration of pathogens, and even intrinsic mechanical stresses arising during tissue growth. Despite the presence of a load-bearing cell wall that separates cells from the environment and fixes their location within a tissue, plant cells are responsive to mechanical stimuli. However, several questions remain unanswered around how mechanical stimuli are perceived and translated into cellular responses. Here we establish a system enabling application of quantifiable localized mechanical stress while simultaneously capturing cellular responses with high spatio-temporal resolution using confocal imaging. We show that this system enables estimation of locally applied pressure and provides access to the temporal and spatial details of subcellular events in intact living tissues upon touch. We propose that observing these subcellular events at high spatiotemporal resolution and linking their dynamics to the intensity of mechanical stimuli may uncover the molecular mechanisms underlying plant cell responses to touch.

plant biology↗

Cell Geometry and Junction Arrangement Define the Mechanical Robustness of Plant Tissues

Plant tissues are composed of immobile, pressurised cells that must maintain structural integrity under diverse environmental loads. Unlike animal tissues, which adapt through cellular rearrangement, plants must achieve mechanical robustness through the geometric configuration of their cellular networks. In this study, we establish a mechanistic link between cell geometry and mechanical resilience by integrating multilayer 3D mechanical simulations with cellular-resolution image analysis across a phylogenetically diverse panel of plant species, including Zea mays, Tradescantia zebrina, and Arabidopsis thaliana. We identify three-way junctions as fundamental mechanical elements that function as flexible hinges enabling mechanical strain accommodation in plant tissues. In contrast, four-way junctions are significantly stiffer and lack this strain-absorbing mechanism, providing a mechanical rationale for their biological avoidance. We also find that tissue material properties and strain response depend on edge lengths, cell layer, the degree of hexagonal shape, and turgor pressure response. These findings reveal the mechanism by which cell division patterns can actively tune tissue resilience to mechanical stress. This work provides new insights into the evolution of different cell shapes and offers clear principles for bio-inspired material science and tissue engineering.

plant biology↗

A dynein-driven nucleokinesis program enables neural crest migration through confined tissues in vivo

Cell migration through confined tissue environments requires precise coordination between the nucleus and the cytoskeleton. Translocation of the nucleus through small pores is a limiting step in confined cell migration. Work in immune and cancer cells has illuminated a key role for actin polymerization and Myosin-II contractility in facilitating nucleus translocation through confinement. However, the microtubule cytoskeleton can also adaptively stabilise under confinement. Microtubule-dependent nucleokinesis constitutes an evolutionarily conserved cellular program ensuring directional nucleus translocation by deployment of cytoplasmic dynein motors. However, whether microtubule-driven nucleokinesis can operate during cell migration through confinement remained so far unexplored. The zebrafish neural crest (NC) provides a unique in vivo system to address this question. NC cells are a highly migratory, multipotent precursors of the vertebrate peripheral nervous system, which traverse diverse environments along the anterior-posterior axis of the embryo. In the head, cranial neural crest (cNC) cells migrate through loosely organised tissues, whereas trunk neural crest (tNC) cells navigate narrowly confined tissue spaces. Here, we show that tNC cells engage a microtubule motor-driven program of nuclear deformation and translocation during confined migration in vivo. Under confinement, tNC cells reorganize their microtubules from a perinuclear meshwork into a polarized, centrosome-associated bundle positioned ahead of the nucleus. Laser ablation reveals that this microtubule array actively pulls and deforms the nucleus. Using pharmacological and genetic approaches, we discover that dynein-dependent pulling forces enable nucleus translocation through confinement. Strikingly, this process occurs independently of Rho/ROCK/myosin II-mediated contractility, revealing a novel neuronal-like mode of nucleokinesis operating in confined tissue environments. Together, our findings uncover a conserved nuclear translocation program deployed during neural crest migration and suggest that microtubule-based nucleokinesis represents a fundamental strategy for navigating tissue confinement in vivo during both central and peripheral nervous system development.

cell biology↗

Notch mediated lateral inhibition is shaped by morphological differences to reinforce bias toward signal-sending or receiving roles

During neurogenesis, neuroblasts are selected from proneural-competent cells through lateral inhibition, a process controlled by the evolutionarily conserved Notch signalling pathway. By tracking transcription from Notch-target genes and cell morphologies in real time, we discovered that the presumptive neuroblast never initiates target-gene transcription. This implies a pre-existing bias directs Notch signalling. The bias correlates with a heterogeneity in apical cell areas which is further reinforced during neuroblast selection. Additionally, the length and duration of neuroblast-neighbour cell contacts predict the likelihood of transcription. Using mathematical modelling we show that lateral inhibition seeded with subtle morphological differences can bias cells toward signal-sending or receiving roles before transcriptional feedback occurs. Notch activation further alters apical cell area, reinforcing the initial bias. We propose that signalling and cell mechanics work together to ensure the robust selection of a single neural precursor.

developmental biology↗

HP1β and densely packed chromatin form separate microdomains in mouse ES cells, which are reconfigured upon exit from naive pluripotency

Heterochromatin proteins play a key role in establishing local chromatin structure to control the transcription of target genes. Here we uncover a surprising segregation between regions of high DNA- and high heterochromatin protein 1{beta} (HP1{beta})-density in mouse ES cells. DNA-low/HP1{beta}-high foci retain freely diffusing HP1{beta}, and form via condensation through a multitude of weak interactions on top of HP1{beta} that is bound stably to chromatin. DNA-high/HP1{beta}-low foci exclude freely diffusing HP1{beta} and display reduced chromatin mobility, suggesting a higher degree of chromatin self-interaction and a more repressive environment. Finally, the two types of environments are intertwined in DNA-high/HP1{beta}-high foci, where HP1{beta} maintains heterochromatin in a more compact yet dynamic chromatin state. During the exit from naive pluripotency HP1{beta} is lost from regions of high DNA density as cells transition through the formative state, which might facilitate the reconfiguration of genome structure accompanying a change in cell state that we observed previously. Subsequently, as cells enter primed pluripotency, canonical heterochromatin is established.

molecular biology↗

Mechanical stress orients stomata division to form tissue scale alignments.

The last stomatal division aligns with the leafs main axis in many species [1]. Understanding how cellular events such as these are coordinated across organ scales remains a challenge in developmental biology. In Arabidopsis, polarised proteins guide the asymmetric divisions in the early stomatal lineage. These proteins show organ scale alignment and may be sensitive to mechanical stress [2]. In contrast, what determines the orientation and alignment of the critical final division is unknown [3]. Here we use an artificial system where every cell adopts the fate of a stomata pore [4] making it easy to visualise their alignment. Combining this system with simultaneous time-lapse imaging on both sides of the cotyledon we are able to compare the stomatal orientation relative to the organ axis, the cell major axis, and the principal directions of growth. Using finite element modelling on a realistic template enabled us to identify differential growth-derived stress patterns as a factor coordinating stomata division at the organ scale. Mechanical perturbation confirmed the influence of tensile stress on stomata division orientation. Through this study, we have identified a mechanism that can explain this nearly century-old observation.

plant biology↗

Local optogenetic NMYII activation within the zebrafish neural rod results in long-range, asymmetric force propagation

How do cellular forces propagate through tissue to allow large-scale morphogenetic events? To investigate this question, we used an in vivo optogenetic approach to reversibly manipulate actomyosin contractility at depth within the developing zebrafish neural rod. Contractility was induced along the lateral cortices of a small patch of developing neural epithelial progenitor cells, resulting in a shortening of these cells along their mediolateral axis. Imaging the immediate response of surrounding tissue uncovered a long-range, tangential, and elastic tissue deformation along the anterior-posterior axis. Unexpectedly, this was highly asymmetric, propagating in either the anterior or the posterior direction in response to local gradients in optogenetic activation. The degree of epithelialisation did not have a significant impact on the extent of force propagation via lateral cortices. We also uncovered a dynamic oscillatory expansion and contraction of the tissue along the anterior-posterior axis, with wavelength within the range of rhombomere length. Together, this study shows dynamic and wave-like propagation of force along the anterior-posterior axis. It also suggests that cell generated forces are actively propagated over long distances within the tissue, and that local anisotropies in tissue organisation and contractility are sufficient to drive directional force propagation.

developmental biology↗

Single-nucleus transcriptomics resolves differentiation dynamics between shoot stem cells and primary stem.

The shoot apical meristem (SAM), located at the plant apex, is accountable for the formation of above-ground organs such as leaves, stem and flowers. Although transcriptional profiling has elucidated some cell-types observed within stems or flowers, the differentiation transcriptional dynamics from shoot stem cells to multiple cell identities remain unknown. We employed a single-nucleus RNA-sequencing approach to assess the transcriptional heterogeneity and cell differentiation processes within the SAM. By collecting dissected inflorescence meristems, we constructed an inflorescence single-nucleus SAM atlas from Arabidopsis thaliana. Our analysis unveiled regulatory elements for most previously known cell types such as the boundary domain, vasculature, early primordia, epidermis and internal stem cells. We also identified previously unobserved transcriptional profiles, revealing that the stem cortex is defined early within forming primordia. Moreover, trajectory inference analysis allowed us to capture spatial control of S-phase machinery by floral homeotic genes and differentiation gene expression dynamics from internal shoot stem cells toward internal layers such as cortex, cambium, xylem and phloem. The results advance our understanding of the cellular and transcriptional heterogeneity underlying the cell-fate transcriptional dynamics shaping shoot organs and architecture.

developmental biology↗

Vortex light field microscopy: 3D spectral single-molecule imaging with a twist

We introduce vortex light field microscopy (VLFM), a novel method for snapshot 3D spectral single-molecule localization microscopy. Inspired by the azimuthal phase profile of optical vortices, we place an azimuthally oriented prism array immediately after the microlens array in a Fourier light field microscope (FLFM). This innovative arrangement causes the axial position and spectral peak for a point emitter to be encoded in the radial and azimuthal displacement of point-spread-function (PSF) respectively. This enables simultaneous detection of 3D position and emission peak of individual fluorophores with 25 nm spatial precision and 3 nm spectral precision over a 4 m depth of field (DOF). We illustrate the spectral scalability of our method by performing four-color 3D single particle tracking of freely diffusing fluorescent beads, and two-color 3D dSTORM imaging of microtubules and mitochondria in fixed COS-7 cells, without the need for spectrally distinct fluorophores.

biophysics↗

Next-generation ABACUS biosensors reveal cellular ABA dynamics driving root growth at low aerial humidity

The plant hormone abscisic acid (ABA) accumulates under abiotic stress to recast water relations and development. To overcome a lack of high-resolution, sensitive reporters, we developed ABACUS2s, next-generation FRET biosensors for ABA with high affinity, signal-to-noise ratio and orthogonality, that reveal endogenous ABA patterns in Arabidopsis thaliana. We mapped stress-induced ABA dynamics in high-resolution to reveal the cellular basis for local and systemic ABA functions. At reduced foliar humidity, roots cells accumulated ABA in the elongation zone, the site of phloem transported ABA unloading. Phloem ABA and root ABA signalling were both essential to maintain root growth at low humidity. ABA coordinates a robust system to maintain root growth in response to foliar stresses, enabling plants to maintain foraging of deeper soil for water uptake

plant biology↗

Single Molecule Light Field Microscopy

We introduce single molecule light field microscopy (SMLFM), a novel 3D single molecule localization technique that is capable of up to 20 nm isotropic precision across a 6 m depth of field. SMLFM can be readily implemented by installing a refractive microlens array into the conjugate back focal plane of any widefield single molecule localization system. We demonstrate that 3D localization can be performed by post-processing 2D localization data generated by common, widely-used, algorithms. In this work we benchmark the performance of SMLFM and finally showcase its capabilities by imaging fluorescently labeled membranes of fixed eukaryotic cells below the diffraction limit.

biophysics↗

Super-resolving light fields in microscopy: depth from disparity

Single molecule localisation microscopy (SMLM) has opened a new window for imaging fluorescently labelled biological specimens. Common 3D SMLM techniques enable data collection across an axial range of 1 - 5m with high precision. Despite the success of 3D single molecule imaging there is a real need to image larger volumes. Here we demonstrate, through simulation and experiment, the potential of Single Molecule Light Field Microscopy (SMLFM) for extended depth-of-field super-resolution imaging, extracting 3D point source position by measuring the disparity between localizations of a point emitter in multiple perspective views.

bioengineering↗