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PMID: 24397990 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, U.S. Gov't, Non-P.H.S.

The role of mechanical tension on lipid raft dependent PDGF-induced TRPC6 activation.

Biomaterials ·Vol. 35 ·No. 9 ·2014-03-00 ·Pages 2868-77

Lei L, Lu S, Wang Y, Kim T, Mehta D, Wang Y

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.

Keywords
Canonical transient receptor potential 6 (TRPC6) Fluorescence resonance energy transfer (FRET) Intracellular tension Lipid rafts Mechanical microenvironment
MeSH Terms
Animals Becaplermin Caveolae/drug effects,metabolism Diffusion Embryo, Mammalian/cytology Fibroblasts/cytology,drug effects,metabolism Fluorescence Resonance Energy Transfer G(M1) Ganglioside/metabolism Genes, Reporter HEK293 Cells Humans Intracellular Space/drug effects,metabolism Ion Channel Gating/drug effects Membrane Microdomains/metabolism Mice Models, Biological Proto-Oncogene Proteins c-sis/pharmacology Rats Signal Transduction/drug effects Stress, Mechanical TRPC Cation Channels/metabolism TRPC6 Cation Channel Type C Phospholipases/metabolism
Chemicals
Proto-Oncogene Proteins c-sis TRPC Cation Channels TRPC6 Cation Channel TRPC6 protein, human Becaplermin G(M1) Ganglioside Type C Phospholipases
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Lei Lei
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.
Lu Shaoying
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.
Wang Yi
Department of Bioengineering & Beckman Institute for Advanced Science and Technology, University of Illinois, Urbana-Champaign, Urbana, IL 61801, United States.
Kim Taejin
Department of Bioengineering & Beckman Institute for Advanced Science and Technology, University of Illinois, Urbana-Champaign, Urbana, IL 61801, United States.
Mehta Dolly
Department of Pharmacology, College of Medicine, University of Illinois, Chicago, IL 60612, United States.
Wang Yingxiao
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.
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Article Info
Journal
Biomaterials
Abbr.
Biomaterials
ISSN
1878-5905
DOI
10.1016/j.biomaterials.2013.12.030
Email
yiw015@eng.ucsd.edu
Published
2014-03-00
Epub
2014-00-04
Pages
2868-77
Language
English
Region
Netherlands
NLM ID
8100316
PMCID
PMC3925146
Subset
IM
Grants
NHLBI NIH HHS · HL098472 · United States
NHLBI NIH HHS · R01 HL071794 · United States
NHLBI NIH HHS · R01 HL084153 · United States
NHLBI NIH HHS · R01 HL098472 · United States
NHLBI NIH HHS · K02 HL109142 · United States
NHLBI NIH HHS · P01 HL060678 · United States
NIGMS NIH HHS · R21 GM106403 · United States
External Links
PubMed source DOI: 10.1016/j.biomaterials.2013.12.030 PMC Full Text View journal details
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