title: The role of mechanical tension on lipid raft dependent PDGF-induced TRPC6 activation.
abstract: Canonical transient receptor potential channel 6 (TRPC6) can play an important role in governing how cells perceive the surrounding material environment and regulate Ca(2+) signaling. We have designed a TRPC6 reporter based on fluorescence resonance energy transfer (FRET) to visualize the TRPC6-mediated calcium entry and hence TRPC6 activity in live cells with high spatiotemporal resolutions. In mouse embryonic fibroblasts (MEFs), platelet-derived growth factor BB (PDGF) can activate the TRPC6 reporter, mediated by phospholipase C (PLC). This TRPC6 activation occurred mainly at lipid rafts regions of the plasma membrane because disruption of lipid raft/caveolae by methyl-β-cyclodextrin (MβCD) or the expression of dominant-negative caveolin-1 inhibited the TRPC6 activity. Culturing cells on soft materials or releasing the intracellular tension by ML-7 reduced this PDGF-induced activation of TRPC6 without affecting the PDGF-regulated Src or inositol 1,4,5-trisphosphate (IP3) receptor function, suggesting a specific role of mechanical tension in regulating TRPC6. We further showed that the release of intracellular tension had similar effect on the diffusion coefficients of TRPC6 and a raft marker, confirming a strong coupling between TRPC6 and lipid rafts. Therefore, our results suggest that the TRPC6 activation mainly occurs at lipid rafts, which is regulated by the mechanical cues of surrounding materials.
authors.affiliation: Department of Bioengineering & Beckman Institute for Advanced Science and Technology, University of Illinois, Urbana-Champaign, Urbana, IL 61801, United States; Department of Bioengineering & Institute of Engineering in Medicine, University of California, San Diego, La Jolla, CA 92093, United States.
authors.forename: Lei
authors.initials: L
authors.lastname: Lei
authors.affiliation: Department of Bioengineering & Beckman Institute for Advanced Science and Technology, University of Illinois, Urbana-Champaign, Urbana, IL 61801, United States; Department of Bioengineering & Institute of Engineering in Medicine, University of California, San Diego, La Jolla, CA 92093, United States.
authors.forename: Shaoying
authors.initials: S
authors.lastname: Lu
authors.affiliation: Department of Bioengineering & Beckman Institute for Advanced Science and Technology, University of Illinois, Urbana-Champaign, Urbana, IL 61801, United States.
authors.forename: Yi
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Department of Bioengineering & Beckman Institute for Advanced Science and Technology, University of Illinois, Urbana-Champaign, Urbana, IL 61801, United States.
authors.forename: Taejin
authors.initials: T
authors.lastname: Kim
authors.affiliation: Department of Pharmacology, College of Medicine, University of Illinois, Chicago, IL 60612, United States.
authors.forename: Dolly
authors.initials: D
authors.lastname: Mehta
authors.affiliation: Department of Bioengineering & Beckman Institute for Advanced Science and Technology, University of Illinois, Urbana-Champaign, Urbana, IL 61801, United States; Department of Bioengineering & Institute of Engineering in Medicine, University of California, San Diego, La Jolla, CA 92093, United States. Electronic address: yiw015@eng.ucsd.edu.
authors.forename: Yingxiao
authors.initials: Y
authors.lastname: Wang
mesh_terms: D000818:Animals; D000077214:Becaplermin; D021941:Caveolae / Q000187:drug effects / Q000378:metabolism; D004058:Diffusion; D004622:Embryo, Mammalian / Q000166:cytology; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D031541:Fluorescence Resonance Energy Transfer; D005677:G(M1) Ganglioside / Q000378:metabolism; D017930:Genes, Reporter; D057809:HEK293 Cells; D006801:Humans; D042541:Intracellular Space / Q000187:drug effects / Q000378:metabolism; D015640:Ion Channel Gating / Q000187:drug effects*; D021962:Membrane Microdomains / Q000378:metabolism*; D051379:Mice; D008954:Models, Biological; D020574:Proto-Oncogene Proteins c-sis / Q000494:pharmacology*; D051381:Rats; D015398:Signal Transduction / Q000187:drug effects; D013314:Stress, Mechanical; D050052:TRPC Cation Channels / Q000378:metabolism*; D000074287:TRPC6 Cation Channel; D010738:Type C Phospholipases / Q000378:metabolism
chemical_list: D020574:Proto-Oncogene Proteins c-sis; D050052:TRPC Cation Channels; D000074287:TRPC6 Cation Channel; C117032:TRPC6 protein, human; D000077214:Becaplermin; D005677:G(M1) Ganglioside; D010738:Type C Phospholipases
keywords: Canonical transient receptor potential 6 (TRPC6); Fluorescence resonance energy transfer (FRET); Intracellular tension; Lipid rafts; Mechanical microenvironment