title: Selective activation of the MEK-ERK pathway is regulated by mechanical stimuli in forming joints and promotes pericellular matrix formation.
abstract: It is well established that local modification of extracellular matrix (ECM) hyaluronan composition is vital in the regulation of cell behavior. Indeed, the formation of articulating chick joint cavities, which requires mechanical stimuli derived from skeletal movement, is dependent upon the accumulation of an ECM rich in hyaluronan (HA). However, the mechanisms responsible for such precise mechano-dependent regulation of cell behavior and the formation of a HA-rich ECM remain undefined. Here we show that extracellular-regulated kinase 1/2 (ERK1/2) is selectively activated in cells at sites of cavity formation and activity diminished by in ovo immobilization that induces cartilaginous fusion across presumptive joint interzones. In vitro analyses offer mechanistic support for the role of mechanical stimuli in promoting a MEK-dependent activation of ERK1/2. In addition, our direct regulation of ERK1/2 phosphorylation status via modulation of its up-stream "classical cascade" activator either pharmacologically or by transfection with dominant negative or constitutively active Mek confirms the essential role for ERK1/2 activation in the elaboration of HA-rich pericellular matrices. Together, our findings demonstrate that the MEK-ERK pathway, regulated by mechanical stimuli, controls HA-rich matrix assembly. The precision of ERK1/2 activation selectively distinguishing cells at the joint line suggests that it directly contributes to the loss of tissue cohesion essential for generating HA-rich cavities between joint elements during their development.
authors.affiliation: Department of Veterinary Basic Sciences, Royal Veterinary College, Royal College St., London, NW1 0TU, United Kingdom.
authors.forename: Edward R
authors.initials: ER
authors.lastname: Bastow
authors.forename: Katherine J
authors.initials: KJ
authors.lastname: Lamb
authors.forename: Jo C
authors.initials: JC
authors.lastname: Lewthwaite
authors.forename: Anne C
authors.initials: AC
authors.lastname: Osborne
authors.forename: Emma
authors.initials: E
authors.lastname: Kavanagh
authors.forename: Caroline P D
authors.initials: CP
authors.lastname: Wheeler-Jones
authors.forename: Andrew A
authors.initials: AA
authors.lastname: Pitsillides
mesh_terms: D000818:Animals; D002642:Chick Embryo; D003593:Cytoplasm / Q000201:enzymology; D004789:Enzyme Activation; D005109:Extracellular Matrix / Q000502:physiology*; D018960:Hyaluronan Receptors / Q000032:analysis; D006820:Hyaluronic Acid / Q000096:biosynthesis*; D007596:Joints / Q000196:embryology*; D020930:MAP Kinase Kinase Kinases / Q000502:physiology*; D019950:Mitogen-Activated Protein Kinase 1 / Q000502:physiology*; D048052:Mitogen-Activated Protein Kinase 3 / Q000502:physiology*; D014532:Uridine Diphosphate Glucose / Q000378:metabolism
chemical_list: D018960:Hyaluronan Receptors; D006820:Hyaluronic Acid; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D020930:MAP Kinase Kinase Kinases; D014532:Uridine Diphosphate Glucose