Signalling dynamics in embryonic development

In multicellular organisms, cellular behaviour is tightly regulated to allow proper embryonic development and maintenance of adult tissue. A critical component in this control is the communication between cells via signalling pathways, as errors in intercellular communication can induce developmental defects or diseases such as cancer. It has become clear over the last years that signalling is not static but varies in activity over time. Feedback mechanisms present in every signalling pathway lead to diverse dynamic phenotypes, such as transient activation, signal ramping or oscillations, occurring in a cell type- and stage-dependent manner. In cells, such dynamics can exert various functions that allow organisms to develop in a robust and reproducible way. Here, we focus on Erk, Wnt and Notch signalling pathways, which are dynamic in several tissue types and organisms, including the periodic segmentation of vertebrate embryos, and are often dysregulated in cancer. We will discuss how biochemical processes influence their dynamics and how these impact on cellular behaviour within multicellular systems.

Can ECIS Biosensor Technology Be Used to Measure the Cellular Responses of Glioblastoma Stem Cells?

Glioblastoma is considered the most aggressive and lethal form of brain cancer. Glioblastoma tumours are complex, comprising a spectrum of oncogenically transformed cells displaying distinct phenotypes. These can be generated in culture and are called differentiated-glioblastoma cells and glioblastoma stem cells. These cells are phenotypically and functionally distinct, where the stem-like glioblastoma cells give rise to and perpetuate the tumour. Electric cell-substrate impedance sensing (ECIS) is a real-time, label-free, impedance-based method for the analysis of cellular behaviour, based on cellular adhesion. Therefore, we asked the question of whether ECIS was suitable for, and capable of measuring the adhesion of glioblastoma cells. The goal was to identify whether ECIS was capable of measuring glioblastoma cell adhesion, with a particular focus on the glioblastoma stem cells. We reveal that ECIS reliably measures adhesion of the differentiated glioblastoma cells on various array types. We also demonstrate the ability of ECIS to measure the migratory behaviour of differentiated glioblastoma cells onto ECIS electrodes post-ablation. Although the glioblastoma stem cells are adherent, ECIS is substantially less capable at reliably measuring their adhesion, compared with the differentiated counterparts. This means that ECIS has applicability for some glioblastoma cultures but much less utility for weakly adherent stem cell counterparts.

Fabricating a Low-cost and Low-immune Xenogeneic Aortic Valve Bio Prosthesis

BackgroundCardiac valve replacement is the only available treatment for end-stage valvular dysfunction patients. In this treatment, among the available choices of valves, the bio-prosthetic valves are better than the mechanical ones in terms of hemodynamic and infection-resistant properties. However, they tend to fail with time, posing a catastrophic event. This study focuses on fabricating the heart valve to eliminate the flaws of bio-prosthetic valves. MethodsPerfusion-based decellularization method was adapted for decellularisation to the sheep heart. Further, decellularised aortic valves were cross-linked with 0.2% Glutaraldehyde (Group C). ResultsAll valves were tested for biochemical and molecular assays including biomechanical tensile testing. Histology, SEM showed a complete lack of cells with intact matrix for decellularised groups. The fibrin glue coated valves leaflet scaffolds showed remodeling of the cells as per the matrix (plasticity). Characterization studies emphasized the cellular behaviour onto matrigel assay, live-dead assay, and the expression of vWF, glycocalyx lectin. ConclusionsThis study focuses on fabricating a re-endothelialized xenogeneic aortic valve leaflet using cross-linking reaction to mask antigenicity of the host proteins (low-immune humanized) and avoid post-implantation cross-reaction.

High-Resolution Imaging for the Analysis and Reconstruction of 3D Microenvironments for Regenerative Medicine: An Application-Focused Review

The rapid evolution of regenerative medicine and its associated scientific fields, such as tissue engineering, has provided great promise for multiple applications where replacement and regeneration of damaged or lost tissue is required. In order to evaluate and optimise the tissue engineering techniques, visualisation of the material of interest is crucial. This includes monitoring of the cellular behaviour, extracellular matrix composition, scaffold structure, and other crucial elements of biomaterials. Non-invasive visualisation of artificial tissues is important at all stages of development and clinical translation. A variety of preclinical and clinical imaging methods—including confocal multiphoton microscopy, optical coherence tomography, magnetic resonance imaging (MRI), and computed tomography (CT)—have been used for the evaluation of artificial tissues. This review attempts to present the imaging methods available to assess the composition and quality of 3D microenvironments, as well as their integration with human tissues once implanted in the human body. The review provides tissue-specific application examples to demonstrate the applicability of such methods on cardiovascular, musculoskeletal, and neural tissue engineering.

JNK Mediates Differentiation, Cell Polarity and Apoptosis During Amphioxus Development by Regulating Actin Cytoskeleton Dynamics and ERK Signalling

c-Jun N-terminal kinase (JNK) is a multi-functional protein involved in a diverse array of context-dependent processes, including apoptosis, cell cycle regulation, adhesion, and differentiation. It is integral to several signalling cascades, notably downstream of non-canonical Wnt and mitogen activated protein kinase (MAPK) signalling pathways. As such, it is a key regulator of cellular behaviour and patterning during embryonic development across the animal kingdom. The cephalochordate amphioxus is an invertebrate chordate model system straddling the invertebrate to vertebrate transition and is thus ideally suited for comparative studies of morphogenesis. However, next to nothing is known about JNK signalling or cellular processes in this lineage. Pharmacological inhibition of JNK signalling using SP600125 during embryonic development arrests gastrula invagination and causes convergence extension-like defects in axial elongation, particularly of the notochord. Pharynx formation and anterior oral mesoderm derivatives like the preoral pit are also affected. This is accompanied by tissue-specific transcriptional changes, including reduced expression of six3/6 and wnt2 in the notochord, and ectopic wnt11 in neurulating embryos treated at late gastrula stages. Cellular delamination results in accumulation of cells in the gut cavity and a dorsal fin-like protrusion, followed by secondary Caspase-3-mediated apoptosis of polarity-deficient cells, a phenotype only partly rescued by co-culture with the pan-Caspase inhibitor Z-VAD-fmk. Ectopic activation of extracellular signal regulated kinase (ERK) signalling in the neighbours of extruded notochord and neural cells, possibly due to altered adhesive and tensile properties, as well as defects in cellular migration, may explain some phenotypes caused by JNK inhibition. Overall, this study supports conserved functions of JNK signalling in mediating the complex balance between cell survival, apoptosis, differentiation, and cell fate specification during cephalochordate morphogenesis.

Die Endozytose — ein zellulärer Aufnahmeweg mit vielfältigen Funktionen

The plasma membrane harbors a specific set of transmembrane proteins which enable diverse cellular functions such as nutrient uptake, ion homeostasis and cellular signaling. The surface levels of these proteins need to be dynamically regulated to allow for plastic changes in cellular behaviour e. g. upon cell stress or during neuronal communication. Endocytosis is a powerful mechanism for quickly adapting the surface proteome via protein internalization. Here, I discuss how endocytosis contributes to brain function and counteracts cell stress.

Ein Wechselbad für Mikroben: Wachsen unter dynamischen Umweltbedingungen

Microfluidic technologies are increasingly used within microbiology for screening and improved understanding of cellular behaviour. The presented system for dynamic microfluidic single-cell cultivation — dMSCC — offers a new cultivation method for microbial microcolonies at dynamic environments with oscillation between seconds to hours. This tool has a high potential to improve the understanding of cellular behaviour at dynamic environments as omnipresent in nature and lab-scale cultivation systems.

Chlamydomonas reinhardtii exhibits stress memory in the accumulation of triacylglycerols induced by nitrogen deprivation.

Stress memory is a phenomenon whereby exposure to initial stress event influences a response to subsequent stress exposures. Studying stress memory is important to understand the cellular behaviour in dynamic environment, especially nowadays, in times with growing environmental instability. Stress memory has been characterized in vascular plants but its occurrence in non-vascular plant species has been rarely investigated. We hypothesized that stress memory occurs in non-vascular plants in relation to metabolic stress. We sought to test it using accumulation of lipids (triacylglycerols) in model green alga Chlamydomonas reinhardtii subjected to nitrogen deprivation stress as a model system. Here, we established stress memory protocol on C. reinhardtii cells. Using a blend of microscopy and gas chromatography methods, we showed that the cells exposed to recurrent stress show differential accumulation of triacylglycerols on the quantitative level without qualitative changes in lipid composition, comparing to single stress controls. Overall, our results suggest that metabolic stress memory does occur in non-vascular plant C. reinhardtii and provides a starting point to characterize metabolic stress memory mechanism. Due to the commercial potential of algae, our findings are relevant for basic science, as well as industrial production of algae-derived compounds.