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J Anat
2017 Dec 01;2316:823-834. doi: 10.1111/joa.12685.
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Expression and functional proteomic analyses of osteocytes from Xenopus laevis tested under mechanical stress conditions: preliminary observations on an appropriate new animal model.
Bertacchini J
,
Benincasa M
,
Checchi M
,
Cavani F
,
Smargiassi A
,
Ferretti M
,
Palumbo C
.
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Hitherto, the role of the osteocyte as transducer of mechanical stimuli into biological signals is far from settled. In this study, we used an appropriate model represented by the cortex of Xenopus laevis long bonediaphysis lacking (unlike the mammalian one) of vascular structures and containing only osteocytes inside the bone matrix. These structural features allow any change of protein profile that might be observed upon different experimental conditions, such as bone adaptation to stress/mechanical loading, to be ascribed specifically to osteocytes. The study was conducted by combining ultrastructural observations and two-dimensional electrophoresis for proteomic analysis. The osteocyte population was extracted from long bones of lower limbs of amphibian skeletons after different protocols (free and forced swimming). The experiments were performed on 210 frogs subdivided into five trials, each including free swimming frogs (controls) and frogs submitted to forced swimming (stressed). The stressed groups were obliged to swim (on movable spheres covering the bottom of a pool on a vibrating plate) continuously for 8 h, and killed 24 h later along with the control groups. Long bones free of soft tissues (periosteum, endosteum and bone marrow), as well as muscles of posterior limbs, were processed and analyzed for proteins differentially expressed or phosphorylated between the two sample groups. The comparative analysis showed that protein phosphorylation profiles differ between control and stressed groups. In particular, we found in long bones of stressed samples that both Erk1/2 and Akt are hyperphosphorylated; moreover, the different phosphorylation of putative Akt substrates (recognized by specific Akt phosphosubstrates-antibody) in stressed vs. control samples clearly demonstrated that Akt signaling is boosted by forced swimming (leading to an increase of mechanical stress) of amphibian long bones. In parallel, we found in posteriorlimb muscles that the expression of heat shock protein HSP27 and HSP70 stress markers increased upon the forced swimming condition. Because the cortexes of frog long bones are characterized by the presence of only osteocytes, all our results establish the suitability of the X. laevis animal model to study the bone response to stress conditions mediated by this cell type and pave the way for further analysis of the signaling pathways involved in these signal transduction mechanisms.
Almeida,
Wnt proteins prevent apoptosis of both uncommitted osteoblast progenitors and differentiated osteoblasts by beta-catenin-dependent and -independent signaling cascades involving Src/ERK and phosphatidylinositol 3-kinase/AKT.
2005, Pubmed
Almeida,
Wnt proteins prevent apoptosis of both uncommitted osteoblast progenitors and differentiated osteoblasts by beta-catenin-dependent and -independent signaling cascades involving Src/ERK and phosphatidylinositol 3-kinase/AKT.
2005,
Pubmed
Basso,
Effect of simulated weightlessness on osteoprogenitor cell number and proliferation in young and adult rats.
2005,
Pubmed
Bellido,
Osteocyte-driven bone remodeling.
2014,
Pubmed
Bertacchini,
Feedbacks and adaptive capabilities of the PI3K/Akt/mTOR axis in acute myeloid leukemia revealed by pathway selective inhibition and phosphoproteome analysis.
2014,
Pubmed
Bertacchini,
The protein kinase Akt/PKB regulates both prelamin A degradation and Lmna gene expression.
2013,
Pubmed
Bonewald,
Mechanosensation and Transduction in Osteocytes.
2006,
Pubmed
Bonewald,
The amazing osteocyte.
2011,
Pubmed
Bonewald,
Osteocytes as dynamic multifunctional cells.
2007,
Pubmed
Buie,
Reduced bone mass accrual in swim-trained prepubertal mice.
2010,
Pubmed
Cao,
Comparative morphology of the osteocyte lacunocanalicular system in various vertebrates.
2011,
Pubmed
,
Xenbase
Cenni,
Lamin A Ser404 is a nuclear target of Akt phosphorylation in C2C12 cells.
2008,
Pubmed
Cowin,
Mechanosensation and fluid transport in living bone.
2002,
Pubmed
Delgado-Calle,
Control of Bone Anabolism in Response to Mechanical Loading and PTH by Distinct Mechanisms Downstream of the PTH Receptor.
2017,
Pubmed
Duncan,
Mechanotransduction and the functional response of bone to mechanical strain.
1995,
Pubmed
Frost,
Bone "mass" and the "mechanostat": a proposal.
1987,
Pubmed
Garman,
Small oscillatory accelerations, independent of matrix deformations, increase osteoblast activity and enhance bone morphology.
2007,
Pubmed
Gennari,
Appropriate models for novel osteoporosis drug discovery and future perspectives.
2015,
Pubmed
Going,
Exercise and Bone Macro-architecture: Is Childhood a Window of Opportunity for Osteoporosis Prevention?
2010,
Pubmed
Govey,
Integrative transcriptomic and proteomic analysis of osteocytic cells exposed to fluid flow reveals novel mechano-sensitive signaling pathways.
2014,
Pubmed
Guadalupe-Grau,
Exercise and bone mass in adults.
2009,
Pubmed
Hong,
Morphological and proteomic analysis of early stage of osteoblast differentiation in osteoblastic progenitor cells.
2010,
Pubmed
Hughes-Fulford,
Reduction of anabolic signals and alteration of osteoblast nuclear morphology in microgravity.
2006,
Pubmed
Klein-Nulend,
Mechanosensation and transduction in osteocytes.
2013,
Pubmed
Kular,
An overview of the regulation of bone remodelling at the cellular level.
2012,
Pubmed
Kumei,
Microgravity signal ensnarls cell adhesion, cytoskeleton, and matrix proteins of rat osteoblasts: osteopontin, CD44, osteonectin, and alpha-tubulin.
2006,
Pubmed
Lammi,
Proteomic analysis of cartilage- and bone-associated samples.
2006,
Pubmed
Lara-Castillo,
In vivo mechanical loading rapidly activates β-catenin signaling in osteocytes through a prostaglandin mediated mechanism.
2015,
Pubmed
Lee,
Proteomics approaches for the studies of bone metabolism.
2014,
Pubmed
Li,
Proteomics based detection of differentially expressed proteins in human osteoblasts subjected to mechanical stress.
2013,
Pubmed
Li,
An integrated proteomics analysis of bone tissues in response to mechanical stimulation.
2011,
Pubmed
Lozupone,
Intermittent compressive load stimulates osteogenesis and improves osteocyte viability in bones cultured "in vitro".
1996,
Pubmed
Maraldi,
Phosphoinositidase C isozymes in SaOS-2 cells: immunocytochemical detection in nuclear and cytoplasmic compartments.
1993,
Pubmed
Marotti,
The osteocyte as a wiring transmission system.
2000,
Pubmed
Marotti,
A quantitative evaluation of osteoblast-osteocyte relationships on growing endosteal surface of rabbit tibiae.
1992,
Pubmed
Marotti,
The structure of bone tissues and the cellular control of their deposition.
1996,
Pubmed
Marotti,
The mechanism of transduction of mechanical strains into biological signals at the bone cellular level.
2007,
Pubmed
Maycas,
PTHrP-Derived Peptides Restore Bone Mass and Strength in Diabetic Mice: Additive Effect of Mechanical Loading.
2017,
Pubmed
McNamara,
Attachment of osteocyte cell processes to the bone matrix.
2009,
Pubmed
Nakahama,
Cellular communications in bone homeostasis and repair.
2010,
Pubmed
Nakashima,
Evidence for osteocyte regulation of bone homeostasis through RANKL expression.
2011,
Pubmed
Palazzini,
Stromal cell structure and relationships in perimedullary spaces of chick embryo shaft bones.
1998,
Pubmed
Palumbo,
Morphological study of intercellular junctions during osteocyte differentiation.
1990,
Pubmed
Palumbo,
Osteocyte-osteoclast morphological relationships and the putative role of osteocytes in bone remodeling.
2001,
Pubmed
Pastorelli,
Proteome analysis for the identification of in vivo estrogen-regulated proteins in bone.
2005,
Pubmed
Poole,
Sclerostin is a delayed secreted product of osteocytes that inhibits bone formation.
2005,
Pubmed
Robling,
Mechanical stimulation of bone in vivo reduces osteocyte expression of Sost/sclerostin.
2008,
Pubmed
Rubinacci,
Bone as an ion exchange system: evidence for a link between mechanotransduction and metabolic needs.
2002,
Pubmed
Rubinacci,
Osteocyte-bone lining cell system at the origin of steady ionic current in damaged amphibian bone.
1998,
Pubmed
,
Xenbase
Rucci,
Characterization of the osteoblast-like cell phenotype under microgravity conditions in the NASA-approved Rotating Wall Vessel bioreactor (RWV).
2002,
Pubmed
Rucci,
Modeled microgravity stimulates osteoclastogenesis and bone resorption by increasing osteoblast RANKL/OPG ratio.
2007,
Pubmed
Ruggeri,
Electron microscopic visualization of proteoglycans with Alcian Blue.
1975,
Pubmed
Schaffler,
Osteocytes: master orchestrators of bone.
2014,
Pubmed
Shahnazari,
Early response of bone marrow osteoprogenitors to skeletal unloading and sclerostin antibody.
2012,
Pubmed
Simkin,
The effect of swimming activity on bone architecture in growing rats.
1989,
Pubmed
Snyder,
The effects of exercise mode, swimming vs. running, upon bone growth in the rapidly growing female rat.
1992,
Pubmed
Sutherland,
Sclerostin promotes the apoptosis of human osteoblastic cells: a novel regulation of bone formation.
2004,
Pubmed
Swissa-Sivan,
The effect of swimming on bone modeling and composition in young adult rats.
1990,
Pubmed
Swissa-Sivan,
Effect of swimming on bone growth and development in young rats.
1989,
Pubmed
Tamma,
Microgravity during spaceflight directly affects in vitro osteoclastogenesis and bone resorption.
2009,
Pubmed
Thi,
Mechanosensory responses of osteocytes to physiological forces occur along processes and not cell body and require αVβ3 integrin.
2013,
Pubmed
Thompson,
Perlecan/Hspg2 deficiency alters the pericellular space of the lacunocanalicular system surrounding osteocytic processes in cortical bone.
2011,
Pubmed
Thompson,
Mechanical regulation of signaling pathways in bone.
2012,
Pubmed
Thompson,
Exercise-induced HSP27, HSP70 and MAPK responses in human skeletal muscle.
2003,
Pubmed
Turner,
Functional determinants of bone structure: beyond Wolff's law of bone transformation.
1992,
Pubmed
Turner,
Homeostatic control of bone structure: an application of feedback theory.
1991,
Pubmed
Wang,
Perlecan-containing pericellular matrix regulates solute transport and mechanosensing within the osteocyte lacunar-canalicular system.
2014,
Pubmed
Wang,
Strain amplification and integrin based signaling in osteocytes.
2008,
Pubmed
Wang,
Evidence for reduced cancellous bone mass in the spontaneously hypertensive rat.
1993,
Pubmed
Warner,
Adaptations in cortical and trabecular bone in response to mechanical loading with and without weight bearing.
2006,
Pubmed
Wijeratne,
Single molecule force measurements of perlecan/HSPG2: A key component of the osteocyte pericellular matrix.
2016,
Pubmed
Xu,
Mechanical stress-induced heat shock protein 70 expression in vascular smooth muscle cells is regulated by Rac and Ras small G proteins but not mitogen-activated protein kinases.
2000,
Pubmed
Zhang,
Label-free quantitative proteome analysis of skeletal tissues under mechanical load.
2009,
Pubmed