Dataset Information

Neutrophil morphology and migration are affected by substrate elasticity.

ABSTRACT: To reach sites of inflammation, neutrophils execute a series of adhesion and migration events that include transmigration through the vascular endothelium and chemotaxis through the vicinal extracellular matrix until contact is made with the point of injury or infection. These in vivo microenvironments differ in their mechanical properties. Using polyacrylamide gels of physiologically relevant elasticity in the range of 5 to 100 kPa and coated with fibronectin, we tested how neutrophil adhesion, spreading, and migration were affected by substrate stiffness. Neutrophils on the softest gels showed only small changes in spread area, whereas on the stiffest gels they showed a 3-fold increase. During adhesion and migration, the magnitudes of the distortions induced in the gel substrate were independent of substrate stiffness, corresponding to the generation of significantly larger traction stresses on the stiffer gels. Cells migrated more slowly but more persistently on stiffer substrates, which resulted in neutrophils moving greater distances over time despite their slower speeds. The largest tractions were localized to the posterior of migrating neutrophils and were independent of substrate stiffness. Finally, the phosphatidylinositol 3-kinase inhibitor LY294002 obviated the ability to sense substrate stiffness, suggesting that phosphatidylinositol 3-kinase plays a mechanistic role in neutrophil mechanosensing.

SUBMITTER: Oakes PW

PROVIDER: S-EPMC2727411 | biostudies-literature | 2009 Aug

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Neutrophil morphology and migration are affected by substrate elasticity.

Oakes Patrick W PW Patel Dipan C DC Morin Nicole A NA Zitterbart Daniel P DP Fabry Ben B Reichner Jonathan S JS Tang Jay X JX

Blood 20090602 7

To reach sites of inflammation, neutrophils execute a series of adhesion and migration events that include transmigration through the vascular endothelium and chemotaxis through the vicinal extracellular matrix until contact is made with the point of injury or infection. These in vivo microenvironments differ in their mechanical properties. Using polyacrylamide gels of physiologically relevant elasticity in the range of 5 to 100 kPa and coated with fibronectin, we tested how neutrophil adhesion, ...[more]

PMID: 19491394

Similar Datasets

Project description:Urbanization alters the environment along many dimensions, including changes to structural habitat and thermal regimes. These can present challenges, but may also provide suitable habitat for certain species. Importantly, the functional implications of these habitat shifts can be assessed through the morphology-performance-fitness paradigm, though these relationships are complicated by interactions among habitat choice, other abiotic factors, and morphology across scales (i.e., micromorphology and gross anatomy). The common wall lizard (Podarcis muralis) is one example of a cosmopolitan and successful urban colonizer. Quantifying both shifts in morphology over time and morphology-performance relationships under various ecological contexts can provide insight into the success of species in a novel environment. To examine how morphological variation influences performance, we measured seven gross morphological characteristics and utilized scanning electron microscopy to obtain high-resolution images of a claw from individuals living in established populations in Cincinnati, Ohio, USA. We used a geometric morphometric approach to describe variation in claw shape and then compared the claws of contemporary lizards to those of museum specimens collected approximately 40 years ago, finding that claw morphology has not shifted over this time. We then performed laboratory experiments to measure the clinging and climbing performance of lizards on materials that mimic ecologically relevant substrates. Each individual was tested for climbing performance on two substrates (cork and turf) and clinging performance on three substrates (cork, turf, and sandpaper) and at two temperatures (24ºC and 34ºC). Clinging performance was temperature insensitive, but determined by substrate-specific interactions between body dimensions and claw morphology. Conversely, the main determinant of climbing performance was temperature, though lizards with more elongate claws, as described by the primary axis of variation in claw morphology, climbed faster. Additionally, we found strong evidence for within-individual trade-offs between performance measures such that individuals who are better at clinging are worse at climbing and vice versa. These results elucidate the complex interactions shaping organismal performance in different contexts and may provide insight into how certain species are able to colonize novel urban environments.

Project description:Microglia-the brain's primary immune cells-exert a tightly regulated cascade of pro- and anti-inflammatory effects upon brain pathology, either promoting regeneration or neurodegeneration. Therefore, harnessing microglia emerges as a potential therapeutic concept in neurological research. Recent studies suggest that-besides being affected by chemokines and cytokines-various cell entities in the brain relevantly respond to the mechanical properties of their microenvironment. For example, we lately reported considerable effects of elasticity on neural stem cells, regarding quiescence and differentiation potential. However, the effects of elasticity on microglia remain to be explored.Under the hypothesis that the elasticity of the microenvironment affects key characteristics and functions of microglia, we established an in vitro model of primary rat microglia grown in a polydimethylsiloxane (PDMS) elastomer-based cell culture system. This way, we simulated the brain's physiological elasticity range and compared it to supraphysiological stiffer PDMS controls. We assessed functional parameters of microglia under "resting" conditions, as well as when polarized towards a pro-inflammatory phenotype (M1) by lipopolysaccharide (LPS), or an anti-inflammatory phenotype (M2) by interleukin-4 (IL-4). Microglia viability was unimpaired on soft substrates, but we found various significant effects with a more than two-fold increase in microglia proliferation on soft substrate elasticities mimicking the brain (relative to PDMS controls). Furthermore, soft substrates promoted the expression of the activation marker vimentin in microglia. Moreover, the M2-marker CD206 was upregulated in parallel to an increase in the secretion of Insulin-Like Growth Factor-1 (IGF-1). The upregulation of CD206 was abolished by blockage of stretch-dependent chloride channels. Our data suggest that the cultivation of microglia on substrates of brain-like elasticity promotes a basic anti-inflammatory activation state via stretch-dependent chloride channels. The results highlight the significance of the omnipresent but mostly overlooked mechanobiological effects exerted on microglia and contribute to a better understanding of the complex spatial and temporal interactions between microglia, neural stem cells, and glia, in health and disease.

Dataset Information

Neutrophil morphology and migration are affected by substrate elasticity.

Publications

Neutrophil morphology and migration are affected by substrate elasticity.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets