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1.
Light and electron microscopic analyses of the high deformability of adhesive toe pads in White's tree frog,Litoria caerulea
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White's tree frog (Litoria caerulea) has large, adhesive toe pads that are among the softest of all known biological structures. To explore the morphological basis for the physical properties of the toe pads, the internal microstructure of the toe pads in L. caerulea was examined using both light and transmission electron microscopy. Three design elements that are distinct from other areas of skin were observed. First, the keratinocytes comprising the adhesive surface of the toe pad all contained keratin filament bundles (tonofibrils) exhibiting structural anisotropy. Specifically, the curved conformation of the hierarchical (branching) tonofibrils was characterized by the formation of anastomoses consisting of tonofibrils beneath the adhesive cell surface and stem keratin filament bundles concentrated in the lower‐middle part of the dorsal‐side of adhesive cells. Second, the cytoplasm of keratinocytes in the most superficial cell layer contained glycoproteins (stained by periodic acid/Schiff reagent) that are considered to confer high viscoelasticity. Third, the dermis contained large lymph spaces interspersed with elastic fibers and collagen fibers, which were relatively sparsely distributed compared to the dorsal skin of the toe pads. The profiles of these structures were easily deformed by the slight application of pressure. These findings reaffirmed that the unique internal architecture of the toe pads in L. caerulea contributed to their remarkable softness and high deformability, which in turn increased the contact area and provided improved adaptability to the local topography of natural surfaces. J. Morphol. 277:1509–1516, 2016. © 2016 Wiley Periodicals, Inc. 相似文献
2.
Pablo Perez Goodwyn Andrei Peressadko Heinz Schwarz Victoria Kastner Stanislav Gorb 《Journal of comparative physiology. A, Neuroethology, sensory, neural, and behavioral physiology》2006,192(11):1233-1243
The morphology, ultrastrucure, effective elastic modulus, and adhesive properties of two different smooth-type attachment pads were studied in two orthopteran species. Tettigonia viridissima (Ensifera) and Locusta migratoria (Caelifera) have a similar structural organization of their attachment pads. They both possess a flexible exocuticle, where the cuticular fibrils are fused into relatively large rods oriented at an angle to the surface. The compliant material of the pad contributes to the contact formation with the substrate. However, the pad material structure was found to be different in these two species. L. migratoria pads bear a thick sub-superficial layer, as well as a higher density of rods. The indentation experiments showed a higher effective elastic modulus and a lower work of adhesion for L. migratoria pads. When the indentations were made at different depths, a higher effective elastic modulus was revealed at lower indentation depths in both species. This effect is explained by the higher stiffness of the superficial pad layer. The obtained results demonstrate a clear correlation between density of the fibres, thickness of the superficial layer, compliance of the pad, and its adhesive properties. Such material structures and properties may be dependent on the preferred environment of each species. 相似文献
3.
Structural correlates of increased adhesive efficiency with adult size in the toe pads of hylid tree frogs 总被引:2,自引:0,他引:2
Joanna M. Smith W. Jon. P. Barnes J. Roger Downie Graeme D. Ruxton 《Journal of comparative physiology. A, Neuroethology, sensory, neural, and behavioral physiology》2006,192(11):1193-1204
Tree frogs are able to climb smooth, vertical substrates using specialised toe pads which adhere via an area-based wet adhesive mechanism. Although the link between pads and arboreality in frogs is well-established, few studies have investigated the influence of morphology on adhesion. Trinidadian tree frogs from the genus Hyla are geometrically similar. There is a tendency towards comparatively reduced mass in larger species, but toe pad area increases as expected with isometry. As adhesion is area-dependent, forces are affected directly by the increase in mass relative to pad area, and there is a decrease in the ability of larger species to adhere to smooth rotation platforms. However, there is an increase in force per unit area that suggests larger species have more efficient toe pads. Toe pad structure is very similar though there are variations in the details of a number of features. Crucially, although differences in morphology appeared small they had demonstrable effects on adhesive efficiency of the pads. Epithelial cell area correlates positively with frog length and adhesive efficiency, related features of cell density and intercellular channel length correlate negatively. These findings are discussed in relation to the different forces involved in the tree frogs’ wet adhesive system. 相似文献
4.
W. Jon. P. Barnes Martina Baum Henrik Peisker Stanislav N. Gorb 《Journal of morphology》2013,274(12):1384-1396
Cryo‐scanning electron microscopy (cryo‐SEM) and atomic force microscopy (AFM) offer new avenues for the study of the morphology of tree frog adhesive toe pads. Using these techniques, we compare toe pad microstructure in two distantly related species of tree frog, Litoria caerulea, White (Hylidae) and Rhacophorus prominanus, Smith (Rhacophoridae), in which the toe pads are considered to be convergent. AFM demonstrates the extraordinary similarity of both surface microstructures (largely hexagonal epithelial cells surrounded by deep channels) and nanostructures (an array of nanopillars, ca. 350 nm in diameter, all with a small dimple at the apex). The cryo‐SEM studies examined the distribution of the fibrillar cytoskeleton within the different layers of the stratified toe pad epithelium, demonstrating that the cytoskeletal elements (keratin tonofilaments) that lie at an angle to the surface are relatively poorly developed in L. caerulea, clearly so in comparison to R. prominanus. Cryo‐SEM also enabled the visualization of the fluid layer that is critical to a toe pad's adhesive function. This was achieved by examination of the frozen fluid residues left behind after removal of a toe within the cryo‐SEM's experimental chamber. Such ‘toeprints’ demonstrated the presence of a wedge of fluid surrounding each toe pad, as well as fluid filling the channels that surround each epithelial cell. Cryo‐SEM was used to examine epithelial cell shape. In a sample of 582 cells, 59.5% were hexagonal, the remainder being mainly pentagonal (23.1%) or heptagonal (16.1%). The distribution of differently‐shaped cells was not random, but was not associated with either pad curvature or the distribution of mucous pores that provide fluid for the frogs' wet adhesion mechanism. Our main finding, the great similarity of toe pad structure in these two species, has important implications for biomimetics, for such convergent evolution suggests a good starting point for attempts to develop adhesives that will function in wet conditions. J. Morphol. 274:1384–1396, 2013. © 2013 Wiley Periodicals, Inc. 相似文献
5.
《Zoology (Jena, Germany)》2014,117(1):7-18
Despite having no obvious anatomical modifications to facilitate movement over land, numerous small fishes from divergent teleost lineages make brief, voluntary terrestrial forays to escape poor aquatic conditions or to pursue terrestrial prey. Once stranded, these fishes produce a coordinated and effective “tail-flip” jumping behavior, wherein lateral flexion of the axial body into a C-shape, followed by contralateral flexion of the body axis, propels the fish into a ballistic flight-path that covers a distance of multiple body lengths. We ask: how do anatomical structures that evolved in one habitat generate effective movement in a novel habitat? Within this context, we hypothesized that the mechanical properties of the axial skeleton play a critical role in producing effective overland movement, and that tail-flip jumping species demonstrate enhanced elastic energy storage through increased body flexural stiffness or increased body curvature, relative to non-jumping species. To test this hypothesis, we derived a model to predict elastic recoil work from the morphology of the vertebral (neural and hemal) spines. From ground reaction force (GRF) measurements and high-speed video, we calculated elastic recoil work, flexural stiffness, and apparent material stiffness of the body for Micropterus salmoides (a non-jumper) and Kryptolebias marmoratus (adept tail-flip jumper). The model predicted no difference between the two species in work stored by the vertebral spines, and GRF data showed that they produce the same magnitude of mass-specific elastic recoil work. Surprisingly, non-jumper M. salmoides has a stiffer body than tail-flip jumper K. marmoratus. Many tail-flip jumping species possess enlarged, fused hypural bones that support the caudal peduncle, which suggests that the localized structures, rather than the entire axial skeleton, may explain differences in terrestrial performance. 相似文献
6.
One step forward: contrasting the effects of Toe clipping and PIT tagging on frog survival and recapture probability
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Murilo Guimarães Décio T. Corrêa Sérgio S. Filho Thiago A. L. Oliveira Paul F. Doherty Jr Ricardo J. Sawaya 《Ecology and evolution》2014,4(8):1480-1490
Amphibians have been declining worldwide and the comprehension of the threats that they face could be improved by using mark–recapture models to estimate vital rates of natural populations. Recently, the consequences of marking amphibians have been under discussion and the effects of toe clipping on survival are debatable, although it is still the most common technique for individually identifying amphibians. The passive integrated transponder (PIT tag) is an alternative technique, but comparisons among marking techniques in free‐ranging populations are still lacking. We compared these two marking techniques using mark–recapture models to estimate apparent survival and recapture probability of a neotropical population of the blacksmith tree frog, Hypsiboas faber. We tested the effects of marking technique and number of toe pads removed while controlling for sex. Survival was similar among groups, although slightly decreased from individuals with one toe pad removed, to individuals with two and three toe pads removed, and finally to PIT‐tagged individuals. No sex differences were detected. Recapture probability slightly increased with the number of toe pads removed and was the lowest for PIT‐tagged individuals. Sex was an important predictor for recapture probability, with males being nearly five times more likely to be recaptured. Potential negative effects of both techniques may include reduced locomotion and high stress levels. We recommend the use of covariates in models to better understand the effects of marking techniques on frogs. Accounting for the effect of the technique on the results should be considered, because most techniques may reduce survival. Based on our results, but also on logistical and cost issues associated with PIT tagging, we suggest the use of toe clipping with anurans like the blacksmith tree frog. 相似文献
7.
Nonlinear, linear and failure properties of articular cartilage and meniscus in opposing contact surfaces are poorly known in tension. Relationships between the tensile properties of articular cartilage and meniscus in contact with each other within knee joints are also not known. In the present study, rectangular samples were prepared from the superficial lateral femoral condyle cartilage and lateral meniscus of bovine knee joints. Tensile tests were carried out with a loading rate of 5 mm/min until the tissue rupture. Nonlinear properties of the toe region, linear properties in larger strains, and failure properties of both tissues were analysed. The strain-dependent tensile modulus of the toe region, Young's modulus of the linear region, ultimate tensile stress and toughness were on average 98.2, 8.3, 4.0 and 1.9 times greater (p<0.05) for meniscus than for articular cartilage. In contrast, the toe region strain, yield strain and failure strain were on average 9.4, 3.1 and 2.3 times greater (p<0.05) for cartilage than for meniscus. There was a significant negative correlation between the strain-dependent tensile moduli of meniscus and articular cartilage samples within the same joints (r=−0.690, p=0.014). In conclusion, the meniscus possesses higher nonlinear and linear elastic stiffness and energy absorption capability before rupture than contacting articular cartilage, while cartilage has longer nonlinear region and can withstand greater strains before failure. These findings point out different load carrying demands that both articular cartilage and meniscus have to fulfil during normal physiological loading activities of knee joints. 相似文献
8.
《Annals of botany》1997,79(5):479-485
The purpose of this paper is to determine the extent to which the physical and mechanical properties of dry and green wood samples are correlated. Samples of green (fresh) sap- and heartwood differing in density (ρ) were removed from the trunk of a black locust (Robinia pseudoacaciaL.) tree 30 years old and measuring 15 m in height. These samples were mechanically tested to determine their Young's elastic modulus (E), proportional (elastic) limit (σp), and modulus of rupture (σR). The Young's elastic modulus of green wood samples increased in magnitude to a limit with increasing cross-sectional area of the sample tested. The values of all three mechanical parameters measured for sapwood samples were consistently lower than those measured for heartwood samples with equivalent cross-sectional areas.Ewas linearly and positively correlated with the σpand σRof heartwood tissue samples. All mechanical properties were highly correlated with the density of green heartwood. Likewise, these properties were highly correlated with the specific gravity of wood samples. Based on these results, it is concluded that either the density of fresh wood or the specific gravity of air-dried wood can be used to estimate the mechanical properties of black locust wood based on simple regression curves in the absence of extensive mechanical tests. 相似文献
9.
W. Jon. P. Barnes Christine Oines Joanna M. Smith 《Journal of comparative physiology. A, Neuroethology, sensory, neural, and behavioral physiology》2006,192(11):1179-1191
This allometric study of adhesion in 15 Trinidadian tree frog species investigates how relationships between length, area and mass limit the ability of adult frog species of different sizes to adhere to inclined and overhanging surfaces. Our experiments show that hylid frogs possess an area-based wet adhesive system in which larger species are lighter than expected from isometry and adhere better than expected from their toe pad area. However, in spite of these adaptations, larger species adhere less well than smaller species. In addition to these adhesive forces, tree frogs also generate significant shear forces that scale with mass, suggesting that they are frictional forces. Toe pads detach by peeling and frogs have strategies to prevent peeling from taking place while they are adhering to surfaces, including orienting themselves head-up on slopes. The scaling of tree frog adhesion is also used to distinguish between different models for adhesion, including classic formulae for capillarity and Stefan adhesion. These classic equations grossly overestimate the adhesive forces that tree frogs produce. More promising are peeling models, designed to predict the pull-off forces of adhesive tape. However, more work is required before we can qualitatively and quantitatively describe the adhesive mechanism of tree frogs. 相似文献
10.
Michael V. Turturro Megan C. Christenson Jeffery C. Larson Daniel A. Young Eric M. Brey Georgia Papavasiliou 《PloS one》2013,8(3)
The spatial presentation of immobilized extracellular matrix (ECM) cues and matrix mechanical properties play an important role in directed and guided cell behavior and neovascularization. The goal of this work was to explore whether gradients of elastic modulus, immobilized matrix metalloproteinase (MMP)-sensitivity, and YRGDS cell adhesion ligands are capable of directing 3D vascular sprout formation in tissue engineered scaffolds. PEGDA hydrogels were engineered with mechanical and biofunctional gradients using perfusion-based frontal photopolymerization (PBFP). Bulk photopolymerized hydrogels with uniform mechanical properties, degradation, and immobilized biofunctionality served as controls. Gradient hydrogels exhibited an 80.4% decrease in elastic modulus and a 56.2% decrease in immobilized YRGDS. PBFP hydrogels also demonstrated gradients in hydrogel degradation with degradation times ranging from 10–12 hours in the more crosslinked regions to 4–6 hours in less crosslinked regions. An in vitro model of neovascularization, composed of co-culture aggregates of endothelial and smooth muscle cells, was used to evaluate the effect of these gradients on vascular sprout formation. Aggregate invasion in gradient hydrogels occurred bi-directionally with sprout alignment observed in the direction parallel to the gradient while control hydrogels with homogeneous properties resulted in uniform invasion. In PBFP gradient hydrogels, aggregate sprout length was found to be twice as long in the direction parallel to the gradient as compared to the perpendicular direction after three weeks in culture. This directionality was found to be more prominent in gradient regions of increased stiffness, crosslinked MMP-sensitive peptide presentation, and immobilized YRGDS concentration. 相似文献
11.
We describe the morphology of toe pads in the Himalayan tree frog Philautus annandalii. These are expanded tips of digits and show modifications of their ventral epidermis for adhesion. The outer cells of toe pad epidermis (TPE) bear surface microstructures (0.7 × 0.2 μm), which are keratinized. Their cytoplasm contains no organelles, but pleomorphic nuclei and mucous granules (0.4–0.5 μm) that glue the keratin filaments. In the intermediate cell layer of TPE, similar keratinized microstructures as in the outer cells are present, so that when the outer layer is shed, it is ready with features for adhesion. These cells contain more keratin than the outer cells. The basal cell layer contains thin keratin bundles and usual cell organelles. The dermis contains mucous‐secreting glands, whose ducts open in the outer epidermal cell layer in channels. The dorsal epidermal cells lack surface microstructures and keratin bundles. Ultrastructural features suggest that toe pads utilize the surface microstructures for adhesion aided by mucus, in which the intermediate cell layer seems to bear the shear stress generated during locomotion. Further, TPE can expand and fit into an increased contact area of the substrate. The long, surface microstructures may also help in mechanical interlocking with rough surfaces on plants. 相似文献
12.
Ruben Mercadé-Prieto Colin R. Thomas Zhibing Zhang 《European biophysics journal : EBJ》2013,42(8):613-620
The elastic modulus of the Baker’s yeast (Saccharomyces cerevisiae) cell wall reported in studies using atomic force microscopy (AFM) is two orders of magnitude lower than that obtained using whole cell compression by micromanipulation. Using finite element modelling, it is shown that Hertz-Sneddon analysis cannot be applied to AFM indentation data for single layer core–shell structures. In addition, the Reissner solution for shallow homogeneous spheres is not appropriate for thick walls such as those of yeast cells. In order to explain yeast compression measurements at different length scales, a double layer wall model is presented considering a soft external layer composed of mannoproteins, and a stiff inner layer of β-glucan fibres and chitin. Under this model, previous AFM studies using sharp indenters provide reasonable estimates of the external layer elastic modulus, while micromanipulation provides the total stiffness of the cell wall. Data from both measurements are combined to estimate the mechanical properties of the inner stiff layer. 相似文献
13.
We describe a high-throughput method capable of quantifying the elastic modulus and drying stress of ex vivo samples of human stratum corneum. Spatially resolved drying deformations in circular tissue samples are measured, azimuthally averaged and fitted with a profile based on a linear elastic model. Our approach enables the comparison of the physical effects of different cleansers. We find that cleansing can cause dramatic changes to the mechanical properties of stratum corneum. In some cases, cleansing can lead to an order of magnitude increase in elastic modulus and drying stress. We expect that these mechanical properties have a direct impact on cracking and chapping skin as well as the milder sensation of perceived tightness often experienced after washing. Mechanical drying studies are also combined with drop wetting studies and pyranine staining experiments. This combination of techniques allows one to establish a multidimensional profile of stratum corneum including stiffness, susceptibility to drying, hydrophilicity and barrier functionality. 相似文献
14.
Neil D. Reeves Constantinos N. Maganaris Nicola Maffulli Joern Rittweger 《Journal of biomechanics》2009,42(7):797-803
Minimising post-operative donor site morbidity is an important consideration when selecting a graft for surgical reconstruction of the torn anterior cruciate ligament (ACL). One of the most common procedures, the bone-patellar tendon-bone (BPTB) graft involves removal of the central third from the tendon. However, it is unknown whether the mechanical properties of the donor site (patellar tendon) recover. The present study investigated the mechanical properties of the human patellar tendon in 12 males (mean±S.D. age: 37±14 years) who had undergone surgical reconstruction of the ACL using a BPTB graft between 1 and 10 years before the study (operated knee; OP). The uninjured contralateral knee served as a control (CTRL). Patellar tendon mechanical properties were assessed in vivo combining dynamometry with ultrasound imaging. Patellar tendon stiffness was calculated from the gradient of the tendon's force–elongation curve. Tendon stiffness was normalised to the tendon's dimensions to obtain the tendon's Young's modulus. Cross-sectional area (CSA) of OP patellar tendons was larger by 21% than CTRL tendons (P<0.01). Patellar tendon stiffness was not significantly different between OP and CTRL tendons, but the Young's modulus was lower by 24% in OP tendons (P<0.01). A compensatory enlargement of the patellar tendon CSA, presumably due to scar tissue formation, enabled a recovery of tendon stiffness in the OP tendons. The newly formed tendon tissue had inferior properties as indicated by the reduced tendon Young's modulus, but it increased to a level that enabled recovery of tendon stiffness. 相似文献
15.
Unique features of diatoms are their intricate cell covers (frustules) made out of hydrated, amorphous silica. The frustule defines and maintains cell shape and protects cells against grazers and pathogens, yet it must allow for cell expansion during growth and division. Other siliceous structures have also evolved in some chain-forming species as means for holding neighboring cells together. Characterization and quantification of mechanical properties of these structures are crucial for the understanding of the relationship between form and function in diatoms, but thus far only a handful of studies have addressed this issue. We conducted micro-indentation experiments, using atomic force microscopy (AFM), to examine local variations in elastic (Young''s) moduli of cells and linking structures in the marine, chain-forming diatom Lithodesmium undulatum. Using a fluorescent tracer that is incorporated into new cell wall components we tested the hypothesis that new siliceous structures differ in elastic modulus from their older counterparts. Results show that the local elastic modulus is a highly dynamic property. Elastic modulus of stained regions was significantly lower than that of unstained regions, suggesting that newly formed cell wall components are generally softer than the ones inherited from the parent cells. This study provides the first evidence of differentiation in local elastic properties in the course of the cell cycle. Hardening of newly formed regions may involve incorporation of additional, possibly organic, material but further studies are needed to elucidate the processes that regulate mechanical properties of the frustule during the cell cycle. 相似文献
16.
Variations in the density and stiffness (Young's elastic modulus)of fresh wood samples drawn from different parts of the threemain trunks of a 32-year-old black locust tree,Robinia pseudoacacia(measuring 19.8 m at its highest point), were studied to determinewhether tree ontogeny can achieve a constant safety factor againstmechanical failure. Based on the properties of isolated woodsamples, the fresh density of sapwood decreased along radialtransects from bark to pith, while that of progressively olderheartwood samples increased, on average, towards the centreof each of the three trunks. Along the same radial transects,the Young's elastic modulus of sap- and heartwood increased.In terms of longitudinal changes in wood properties, mean woodmoduli (averages of sap- and heartwood samples) increased, onaverage, towards the base of each of the three trunks of thetree. However, the mean fresh densities of wood samples increasedtowards the top and the bottom of each trunk and were lowestroughly near trunk mid-length. The mean density-specific stiffness(the quotient of Young's modulus and fresh density) of woodwas thus lower toward the top and the bottom of the trunks andhighest near trunk mid-length. Mean values of fresh wood density-specificstiffness were used to estimate the critical buckling heightsfor sections of the trunks differing in diameter and age. Theseestimates indicated that ontogenetic variation in the physicalproperties and relative amounts of sap- and heartwood in trunkscould maintain a constant factor of safety (approximately equalto 2) as a sapling grows in height and girth into a mature tree.This expectation was supported by data from 16 black locusttrees differing in height and diameter at breast height (DBH). Wood; elastic properties; tree height; biomechanics 相似文献
17.
Micromechanics of smooth adhesive organs in stick insects: pads are mechanically anisotropic and softer towards the adhesive surface 总被引:1,自引:0,他引:1
Ingo Scholz Werner Baumgartner Walter Federle 《Journal of comparative physiology. A, Neuroethology, sensory, neural, and behavioral physiology》2008,194(4):373-384
Animals have evolved adhesive structures on their legs to cling to the substrate during locomotion. Here we characterise the
ultrastructure and mechanical properties of adhesive pads in Carausius morosus (Phasmatodea) using atomic force microscopy (AFM) as well as transmission and scanning electron microscopy (TEM, SEM). The
smooth adhesive arolium has a soft cuticle consisting of principal rods, which branch into finer fibres near the surface.
Indentation experiments showed that the pad material consists of distinct layers with different mechanical properties. The
100–300 nm thick outermost layer consisting of the cuticulin envelope and the epicuticle is extremely soft and resilient (mean
effective Young’s modulus 12 kPa), while the subjacent procuticle is a much stiffer material (mean effective Young’s modulus
625 kPa). AFM contact mode imaging revealed that the cuticle is mechanically anisotropic, which can be explained by its fibrillar
inner structure. We propose that the described layered structure of smooth adhesive pads, consisting of materials decreasing
in stiffness towards the outer surface, represents a superior design to conform and adhere to substrates with roughnesses
at different length scales. This design principle could be easily implemented in technical adhesives, and thus has a potential
to inspire biomimetic applications. 相似文献
18.
Ridim D. Mote Jyoti Yadav Surya Bansi Singh Mahak Tiwari Shinde Laxmikant V Shivprasad Patil Deepa Subramanyam 《The Journal of biological chemistry》2020,295(49):16888
Mouse embryonic stem cells (mESCs) display unique mechanical properties, including low cellular stiffness in contrast to differentiated cells, which are stiffer. We have previously shown that mESCs lacking the clathrin heavy chain (Cltc), an essential component for clathrin-mediated endocytosis (CME), display a loss of pluripotency and an enhanced expression of differentiation markers. However, it is not known whether physical properties such as cellular stiffness also change upon loss of Cltc, similar to what is seen in differentiated cells, and if so, how these altered properties specifically impact pluripotency. Using atomic force microscopy (AFM), we demonstrate that mESCs lacking Cltc display higher Young''s modulus, indicative of greater cellular stiffness, compared with WT mESCs. The increase in stiffness was accompanied by the presence of actin stress fibers and accumulation of the inactive, phosphorylated, actin-binding protein cofilin. Treatment of Cltc knockdown mESCs with actin polymerization inhibitors resulted in a decrease in the Young''s modulus to values similar to those obtained with WT mESCs. However, a rescue in the expression profile of pluripotency factors was not obtained. Additionally, whereas WT mouse embryonic fibroblasts could be reprogrammed to a state of pluripotency, this was inhibited in the absence of Cltc. This indicates that the presence of active CME is essential for the pluripotency of embryonic stem cells. Additionally, whereas physical properties may serve as a simple readout of the cellular state, they may not always faithfully recapitulate the underlying molecular fate. 相似文献
19.
Suei S Seyan R Noguera P Manzi J Plastino J Kreplak L 《Journal of molecular biology》2011,413(3):573-583
The comet motility assay, inspired by Listeria locomotion, has been used extensively as an in vitro model to study the structural and motile properties of the actin cytoskeleton. However, there are no quantitative measurements of the mechanical properties of these actin comets. In this work, we use nanoindentation based on atomic force microscopy to measure the elastic modulus of actin comets grown on 1-μm-diameter beads in an Arp2/3 (actin-related proteins 2 and 3)-complex-dependent fashion in the absence and in the presence of VASP (vasodilator-stimulated phosphoprotein). Recruitment of VASP to the bead surface had no effect on the initial velocity or morphology of the comets. Instead, we observed an improved contact of the comets with the beads and an increased elastic modulus of the comets. The VASP-mediated increase in elastic modulus was dependent on both concentration and ionic strength. In conclusion, we propose that VASP plays a mechanical role in Arp2/3-complex-dependent motility by amplifying the elastic modulus of the thus assembled actin network and, consequently, by strengthening its cohesion for persistent protrusion. 相似文献
20.
While the determination of mechanical properties of a hard scaffold is relatively straightforward, the mechanical testing of a soft tissue scaffold poses significant challenges due in part to its fragility. Here, we report a new approach for characterizing the stiffness and elastic modulus of a soft scaffold through atomic force microscopy (AFM) nanoindentation. Using collagen-chitosan hydrogel scaffolds as model soft tissue scaffolds, we demonstrated the feasibility of using AFM nanoindentation to determine a force curve of a soft tissue scaffold. A mathematical model was developed to ascertain the stiffness and elastic modulus of a scaffold from its force curve obtained under different conditions. The elastic modulus of a collagen-chitosan (80%/20%, v/v) scaffold is found to be 3.69 kPa. The scaffold becomes stiffer if it contains more chitosan. The elastic modulus of a scaffold composed of 70% collagen and 30% chitosan is about 11.6 kPa. Furthermore, the stiffness of the scaffold is found to be altered significantly by extracellular matrix deposited from cells that are grown inside the scaffold. The elastic modulus of collagen-chitosan scaffolds increased from 10.5 kPa on day 3 to 63.4 kPa on day 10 when human foreskin fibroblast cells grew inside the scaffolds. Data acquired from these measurements will offer new insights into understanding cell fate regulation induced by physiochemical cues of tissue scaffolds. 相似文献