Shear-induced damped oscillations in an epithelium depend on actomyosin contraction and E-cadherin cell adhesion.

Shear forces between cells occur during global changes in multicellular organization during morphogenesis and tissue growth, yet how cells sense shear forces and propagate a response across a tissue is unknown. We found that applying exogenous shear at the midline of an epithelium induced a local, short-term deformation near the shear plane, and a long-term collective oscillatory movement across the epithelium that spread from the shear-plane and gradually dampened. Inhibiting actomyosin contraction or E-cadherin trans-cell adhesion blocked oscillations, whereas stabilizing actin filaments prolonged oscillations. Combining these data with a model of epithelium mechanics supports a mechanism involving the generation of a shear-induced mechanical event at the shear plane which is then relayed across the epithelium by actomyosin contraction linked through E-cadherin. This causes an imbalance of forces in the epithelium, which is gradually dissipated through oscillatory cell movements and actin filament turnover to restore the force balance across the epithelium

Mechano-transduction: from molecules to tissues.

External forces play complex roles in cell organization, fate, and homeostasis. Changes in these forces, or how cells respond to them, can result in abnormal embryonic development and diseases in adults. How cells sense and respond to these mechanical stimuli requires an understanding of the biophysical principles that underlie changes in protein conformation and result in alterations in the organization and function of cells and tissues. Here, we discuss mechano-transduction as it applies to protein conformation, cellular organization, and multi-cell (tissue) function

Stable, covalent attachment of laminin to microposts improves the contractility of mouse neonatal cardiomyocytes.

The mechanical output of contracting cardiomyocytes, the muscle cells of the heart, relates to healthy and disease states of the heart. Culturing cardiomyocytes on arrays of elastomeric microposts can enable inexpensive and high-throughput studies of heart disease at the single-cell level. However, cardiomyocytes weakly adhere to these microposts, which limits the possibility of using biomechanical assays of single cardiomyocytes to study heart disease. We hypothesized that a stable covalent attachment of laminin to the surface of microposts improves cardiomyocyte contractility. We cultured cells on polydimethylsiloxane microposts with laminin covalently bonded with the organosilanes 3-glycidoxypropyltrimethoxysilane and 3-aminopropyltriethoxysilane with glutaraldehyde. We measured displacement of microposts induced by the contractility of mouse neonatal cardiomyocytes, which attach better than mature cardiomyocytes to substrates. We observed time-dependent changes in contractile parameters such as micropost deformation, contractility rates, contraction and relaxation speeds, and the times of contractions. These parameters were affected by the density of laminin on microposts and by the stability of laminin binding to micropost surfaces. Organosilane-mediated binding resulted in higher laminin surface density and laminin binding stability. 3-glycidoxypropyltrimethoxysilane provided the highest laminin density but did not provide stable protein binding with time. Higher surface protein binding stability and strength were observed with 3-aminopropyltriethoxysilane with glutaraldehyde. In cultured cardiomyocytes, contractility rate, contraction speeds, and contraction time increased with higher laminin stability. Given these variations in contractile function, we conclude that binding of laminin to microposts via 3-aminopropyltriethoxysilane with glutaraldehyde improves contractility observed by an increase in beating rate and contraction speed as it occurs during the postnatal maturation of cardiomyocytes. This approach is promising for future studies to mimic in vivo tissue environments

Somatosensory neurons integrate the geometry of skin deformation and mechanotransduction channels to shape touch sensing.

Touch sensation hinges on force transfer across the skin and activation of mechanosensitive ion channels along the somatosensory neurons that invade the skin. This skin-nerve sensory system demands a quantitative model that spans the application of mechanical loads to channel activation. Unlike prior models of the dynamic responses of touch receptor neurons in Caenorhabditis elegans (Eastwood et al., 2015), which substituted a single effective channel for the ensemble along the TRNs, this study integrates body mechanics and the spatial recruitment of the various channels. We demonstrate that this model captures mechanical properties of the worm's body and accurately reproduces neural responses to simple stimuli. It also captures responses to complex stimuli featuring non-trivial spatial patterns, like extended or multiple contacts that could not be addressed otherwise. We illustrate the importance of these effects with new experiments revealing that skin-neuron composites respond to pre-indentation with increased currents rather than adapting to persistent stimulation

Increasing β-catenin/Wnt3A activity levels drive mechanical strain-induced cell cycle progression through mitosis.

Mechanical force and Wnt signaling activate β-catenin-mediated transcription to promote proliferation and tissue expansion. However, it is unknown whether mechanical force and Wnt signaling act independently or synergize to activate β-catenin signaling and cell division. We show that mechanical strain induced Src-dependent phosphorylation of Y654 β-catenin and increased β-catenin-mediated transcription in mammalian MDCK epithelial cells. Under these conditions, cells accumulated in S/G2 (independent of DNA damage) but did not divide. Activating β-catenin through Casein Kinase I inhibition or Wnt3A addition increased β-catenin-mediated transcription and strain-induced accumulation of cells in S/G2. Significantly, only the combination of mechanical strain and Wnt/β-catenin activation triggered cells in S/G2 to divide. These results indicate that strain-induced Src phosphorylation of β-catenin and Wnt-dependent β-catenin stabilization synergize to increase β-catenin-mediated transcription to levels required for mitosis. Thus, local Wnt signaling may fine-tune the effects of global mechanical strain to restrict cell divisions during tissue development and homeostasis

Phase II Archaeological Testing of the Hothouse Structure (18TA314), Talbot County, Maryland

This report contains the results of the Phase II Archaeological Testing of the Wye House Hothouse Structure (18TA314). It is divided into the following sections: Chapter 1: Introduction Chapter 2: Research Design and Methodology Chapter 3: Cultural Context and Historical Background Chapter 4: Previous Archaeological Investigations and Significant Architectural Structures Chapter 5: Archaeology and Interpretations Chapter 6: Conclusions and Recommendations Chapter 1 of this report is an introduction to the Hothouse excavation in May 2012. Included within in this chapter are the dates of fieldwork, laboratory processing and analyses, as well as the identification of key project staff. Chapter 2 of this report details the project’s research design and methodology. Included within this chapter are the method employed during the research process of locating the structures, excavation, pollen sample recovery, and laboratory processing. In addition, this chapter lists the research questions that guided this fieldwork. Chapter 3 of this report details the cultural context and historical background of the Wye House Plantation, particularly surrounding the standing Greenhouse. Included within this chapter is a short history of the Wye House Plantation, scientific gardening, and a contextualization for understanding West African spirit practices. Chapter 4 of this report details reported archaeological excavations at the Wye House Plantation, particularly surrounding the standing Greenhouse. Chapter 5 of this report details the results of archaeological testing conducted at the Hothouse structure in May 2012. Included within this chapter is an account of stratigraphic layers, features, and significant artifacts encountered within individual test units. Also included within this chapter are interpretations of layers, features, and artifacts. Chapter 6 of this report details the conclusions based on the data recovered from these excavations and recommendations for further investigations

Spatial distribution of cell-cell and cell-ECM adhesions regulates force balance while main-taining E-cadherin molecular tension in cell pairs.

Mechanical linkage between cell-cell and cell-extracellular matrix (ECM) adhesions regulates cell shape changes during embryonic development and tissue homoeostasis. We examined how the force balance between cell-cell and cell-ECM adhesions changes with cell spread area and aspect ratio in pairs of MDCK cells. We used ECM micropatterning to drive different cytoskeleton strain energy states and cell-generated traction forces and used a Förster resonance energy transfer tension biosensor to ask whether changes in forces across cell-cell junctions correlated with E-cadherin molecular tension. We found that continuous peripheral ECM adhesions resulted in increased cell-cell and cell-ECM forces with increasing spread area. In contrast, confining ECM adhesions to the distal ends of cell-cell pairs resulted in shorter junction lengths and constant cell-cell forces. Of interest, each cell within a cell pair generated higher strain energies than isolated single cells of the same spread area. Surprisingly, E-cadherin molecular tension remained constant regardless of changes in cell-cell forces and was evenly distributed along cell-cell junctions independent of cell spread area and total traction forces. Taken together, our results showed that cell pairs maintained constant E-cadherin molecular tension and regulated total forces relative to cell spread area and shape but independently of total focal adhesion area

Phase II Archaeological Testing on Schwar’s Row (18AP120), Annapolis, Maryland, 2012

This report is a summary of excavations conducted by Archaeology in Annapolis between May 29 2012 and July 6 2012 on Cornhill Street, Annapolis, Maryland. The report is divided into the following sections: Chapter 1: Introduction Chapter 2: Context and Historical Background Chapter 3: Archaeology and Interpretations Chapter 4: Conclusions and Recommendations Chapter 1 of this report is an introduction to the Cornhill Street excavations in 2012 at two areas designated Schwar’s Row East and Schwar’s Row West. Included within in this chapter are the dates of fieldwork, laboratory processing and analyses, the identification of key project staff, as well as research design and methodology. Chapter 2 of this report details the context and historical background of the properties. Included within this chapter is a short history of the ownership of the structures and the research questions for this investigation. Chapter 3 of this report details the results of archaeological testing of a total of four units at Schwar’s Row East and Schwar’s Row West. Included within this chapter is an account of stratigraphic layers, features, and significant artifacts encountered within individual test units. Also included within this chapter are interpretations of layers, features, and artifacts. Chapter 4 of this report details the conclusions based on the data recovered from these excavations and recommendations for further investigations