Microparticle assembly pathways on lipid membranes

Understanding interactions between microparticles and lipid membranes is of increasing importance, especially for unraveling the influence of microplastics on our health and environment. Here, we study how a short-ranged adhesive force between microparticles and model lipid membranes causes membrane-mediated particle assembly. Using confocal microscopy, we observe the initial particle attachment to the membrane, then particle wrapping, and in rare cases spontaneous membrane tubulation. In the attached state, we measure that the particle mobility decreases by 26%. If multiple particles adhere to the same vesicle, their initial single-particle state determines their interactions and subsequent assembly pathways: 1) attached particles only aggregate when small adhesive vesicles are present in solution, 2) wrapped particles reversibly attract one another by membrane deformation, and 3) a combination of wrapped and attached particles form membrane-mediated dimers, which further assemble into a variety of complex structures. The experimental observation of distinct assembly pathways induced only by a short ranged membrane-particle adhesion, shows that a cellular cytoskeleton or other active components are not required for microparticle aggregation. We suggest that this membrane-mediated microparticle aggregation is a reason behind reported long retention times of polymer microparticles in organisms.Comment: 20 pages, 11 figures (including supporting material

Gastropoden aus paläozoischen Geschieben des Kies-Sand-Rückens in der Laerheide (Landkreis Osnabrück)

Aus altpaläozoischen karbonatischen Gesteinen, die als nordische Geschiebe in den Kame-Ablagerungen der Laerheide am Nordrand der Westfälischen Bucht vorkommen, werden ordovizische und silurische Gastropoden beschrieben. Altpaläozoische Gastropoden sind zwar aus verschiedenen südskandinavischen Kalkstein-Geschieben seit langem bekannt, jedoch sind Bestimmungen und Eingruppierungen innerhalb der modernen Gastropoden-Systematik bislang noch nicht erfolgt, so dass auf diesem Gebiet noch erheblicher Nachholbedarf besteht, wozu hiermit ein erster Beitrag geleistet wird.Glacial erratic boulders from kame sediments of the Laerheide in the northern part of the „Westfälische Bucht“ south of Osnabrück have yielded Ordovician and Silurian gastropods. Although early Palaeozoic gastropods are fairly well known from glacial erratics they lack adequate descriptions and classification within the modern systematic framework which is presented here

Memory improves precision of cell sensing in fluctuating environments

Biological cells are often found to sense their chemical environment near the single-molecule detection limit. Surprisingly, this precision is higher than simple estimates of the fundamental physical limit, hinting towards active sensing strategies. In this work, we analyse the effect of cell memory, e.g. from slow biochemical processes, on the precision of sensing by cell-surface receptors. We derive analytical formulas, which show that memory significantly improves sensing in weakly fluctuating environments. However, surprisingly when memory is adjusted dynamically, the precision is always improved, even in strongly fluctuating environments. In support of this prediction we quantify the directional biases in chemotactic Dictyostelium discoideum cells in a flow chamber with alternating chemical gradients. The strong similarities between cell sensing and control engineering suggest universal problem-solving strategies of living matter

Bovine polledness

The persistent horns are an important trait of speciation for the family Bovidae with complex morphogenesis taking place briefly after birth. The polledness is highly favourable in modern cattle breeding systems but serious animal welfare issues urge for a solution in the production of hornless cattle other than dehorning. Although the dominant inhibition of horn morphogenesis was discovered more than 70 years ago, and the causative mutation was mapped almost 20 years ago, its molecular nature remained unknown. Here, we report allelic heterogeneity of the POLLED locus. First, we mapped the POLLED locus to a ∼381-kb interval in a multi-breed case-control design. Targeted re-sequencing of an enlarged candidate interval (547 kb) in 16 sires with known POLLED genotype did not detect a common allele associated with polled status. In eight sires of Alpine and Scottish origin (four polled versus four horned), we identified a single candidate mutation, a complex 202 bp insertion-deletion event that showed perfect association to the polled phenotype in various European cattle breeds, except Holstein-Friesian. The analysis of the same candidate interval in eight Holsteins identified five candidate variants which segregate as a 260 kb haplotype also perfectly associated with the POLLED gene without recombination or interference with the 202 bp insertion-deletion. We further identified bulls which are progeny tested as homozygous polled but bearing both, 202 bp insertion-deletion and Friesian haplotype. The distribution of genotypes of the two putative POLLED alleles in large semi-random sample (1,261 animals) supports the hypothesis of two independent mutations

Deformability and collision-induced reorientation enhance cell topotaxis in dense microenvironments

In vivo, cells navigate through complex environments filled with obstacles. Recently, the term 'topotaxis' has been introduced for navigation along topographic cues such as obstacle density gradients. Experimental and mathematical efforts have analyzed topotaxis of single cells in pillared grids with pillar density gradients. A previous model based on active Brownian particles has shown that ABPs perform topotaxis, i.e., drift towards lower pillar densities, due to decreased effective persistence lengths at high pillars densities. The ABP model predicted topotactic drifts of up to 1% of the instantaneous speed, whereas drifts of up to 5% have been observed experimentally. We hypothesized that the discrepancy between the ABP and the experimental observations could be in 1) cell deformability, and 2) more complex cell-pillar interactions. Here, we introduce a more detailed model of topotaxis, based on the Cellular Potts model. To model persistent cells we use the Act model, which mimicks actin-polymerization driven motility, and a hybrid CPM-ABP model. Model parameters were fitted to simulate the experimentally found motion of D. discoideum on a flat surface. For starved D. discoideum, both CPM variants predict topotactic drifts closer to the experimental results than the previous ABP model, due to a larger decrease in persistence length. Furthermore, the Act model outperformed the hybrid model in terms of topotactic efficiency, as it shows a larger reduction in effective persistence time in dense pillar grids. Also pillar adhesion can slow down cells and decrease topotaxis. For slow and less persistent vegetative D. discoideum cells, both CPMs predicted a similar small topotactic drift. We conclude that deformable cell volume results in higher topotactic drift compared to ABPs, and that feedback of cell-pillar collisions on cell persistence increases drift only in highly persistent cells

Modulation of mammalian cell behavior by nanoporous glass

The introduction of novel bioactive materials to manipulate living cell behavior is a crucial topic for biomedical research and tissue engineering. Biomaterials or surface patterns that boost specific cell functions can enable innovative new products in cell culture and diagnostics. This study investigates the influence of the intrinsically nano-patterned surface of nanoporous glass membranes on the behavior of mammalian cells. Three different cell lines and primary human mesenchymal stem cells (hMSCs) proliferate readily on nanoporous glass membranes with mean pore sizes between 10 and 124 nm. In both proliferation and mRNA expression experiments, L929 fibroblasts show a distinct trend toward mean pore sizes >80 nm. For primary hMSCs, excellent proliferation is observed on all nanoporous surfaces. hMSCs on samples with 17 nm pore size display increased expression of COL10, COL2A1, and SOX9, especially during the first two weeks of culture. In the upside down culture, SK-MEL-28 cells on nanoporous glass resist the gravitational force and proliferate well in contrast to cells on flat references. The effect of paclitaxel treatment of MDA-MB-321 breast cancer cells is already visible after 48 h on nanoporous membranes and strongly pronounced in comparison to reference samples, underlining the material's potential for functional drug screening

Contact-controlled amoeboid motility induces dynamic cell trapping in 3D-microstructured surfaces.

On flat substrates, several cell types exhibit amoeboid migration, which is characterized by restless stochastic successions of pseudopod protrusions. The orientation and frequency of new membrane protrusions characterize efficient search modes, which can respond to external chemical stimuli as observed during chemotaxis in amoebae. To quantify the influence of mechanical stimuli induced by surface topography on the migration modes of the amoeboid model organism Dictyostelium discoideum, we apply high resolution motion analysis in microfabricated pillar arrays of defined density and geometry. Cell motion is analyzed by a two-state motility-model, distinguishing directed cellular runs from phases of isotropic migration that are characterized by randomly oriented cellular protrusions. Cells lacking myosin II or cells deprived of microtubules show significantly different behavior concerning migration velocities and migrational angle distribution, without pronounced attraction to pillars. We conclude that microtubules enhance cellular ability to react with external 3D structures. Our experiments on wild-type cells show that the switching from randomly formed pseudopods to a stabilized leading pseudopod is triggered by contact with surface structures. These alternating processes guide cells according to the available surface in their 3D environment, which we observed dynamically and in steady-state situations. As a consequence, cells perform "home-runs" in low-density pillar arrays, crawling from pillar to pillar, with a characteristic dwell time of 75 s. At the boundary between a flat surface and a 3D structured substrate, cells preferentially localize in contact with micropillars, due to the additionally available surface in the microstructured arrays. Such responses of cell motility to microstructures might open new possibilities for cell sorting in surface structured arrays

A two-armed probe for in-cell DEER measurements on proteins

The application of double electron-electron resonance (DEER) with site-directed spin labeling (SDSL) to measure distances in proteins and protein complexes in living cells puts rigorous restraints on the spin-label. The linkage and paramagnetic centers need to resist the reducing conditions of the cell. Rigid attachment of the probe to the protein improves precision of the measured distances. Here, three two-armed GdIII complexes, GdIII-CLaNP13a/b/c were synthesized. Rather than the disulfide linkage of most other CLaNP molecules, a thioether linkage was used to avoid reductive dissociation of the linker. The doubly GdIII labeled N55C/V57C/K147C/T151C variants of T4Lysozyme were measured by 95 GHz DEER. The constructs were measured in vitro, in cell lysate and in Dictyostelium discoideum cells. Measured distances were 4.5 nm, consistent with results from paramagnetic NMR. A narrow distance distribution and typical modulation depth, also in cell, indicate complete and durable labeling and probe rigidity due to the dual attachment sites