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The Journal of Membrane Biology

, Volume 252, Issue 4–5, pp 307–315 | Cite as

Mechanical Stretch Redefines Membrane Gαq–Calcium Signaling Complexes

  • Androniqi Qifti
  • Osama Garwain
  • Suzanne ScarlataEmail author
Article
Part of the following topical collections:
  1. Membrane and Receptor Dynamics

Abstract

Muscle cells are routinely subjected to mechanical stretch but the impact of stretch on the organization of membrane domains is unknown. In this study, we characterize the effect of stretch on GPCR–Gαq protein signaling. Activation of this pathway leads to an increase in intracellular calcium. In muscle cells, GPCR–Gαq signals are enhanced when these proteins are localized in caveolae membrane domains whose curved structure can flatten with stretch. When we statically stretch rat aortic smooth muscle A10 cells by 1–5%, cellular calcium appears unperturbed as indicated by a calcium indicator. However, when we activate the bradykinin type 2 receptor (B2R)/Gαq pathway, we observe a loss in calcium that appears to be mediated through perturbations in calcium-activated stretch receptors. In contrast, if we apply oscillating stretch, calcium levels are enhanced. We tested whether the observed changes in B2R–Gαq calcium signals were caused by stretch-induced disruption of caveolae using a combination of silencing RNA technology and growth conditions. We find that stretch changes the ability of monoclonal caveolin antibodies to bind caveolae indicating a change in configuration of the domains. This change is seen by the inability of cells to survive stretch cycles when the level of caveolae is significantly reduced. Our studies show that the effect of calcium signals by mechanical stretch is mediated by the type of stretch and the amount of caveolae.

Keywords

Calcium signaling G proteins Mechanical stretch Caveolae 

Notes

Acknowledgements

The authors would like to thank Dr. Kristin Billiar (Dept. of Biomedical Engineering, Worcester Polytechnic Institute) for use of his stretch apparatus as well as Zachary Goldblatt for his assistance with the device.

Funding

The authors are grateful for funding from NIH-GM116187 and support from the Richard T. Whitcomb funds.

Compliance with Ethical Standards

Conflict of Interest

The authors declare no conflict of interest.

Ethical Approval

This article does not contain any studies with human participants or animals performed by any of the authors.

References

  1. 1.
    Taggart MJ (2001) Smooth muscle excitation-contraction coupling: a role for caveolae and caveolins? News Physiol Sci 16:61–65PubMedGoogle Scholar
  2. 2.
    Williams TM, Lisanti MP (2004) The caveolin proteins. Genome Biol 5:214CrossRefGoogle Scholar
  3. 3.
    Parton RG, Simons K (2007) The multiple faces of caveolae. Nat Rev Mol Cell Biol 8:185–194CrossRefGoogle Scholar
  4. 4.
    Parton RG, Hanzal-Bayer M, Hancock JF (2006) Biogenesis of caveolae: a structural model for caveolin-induced domain formation. J Cell Sci 119:787–796CrossRefGoogle Scholar
  5. 5.
    Gratton J-P, Bernatchez P, Sessa WC (2004) Caveolae and caveolins in the cardiovascular system. Circ Res 94:1408–1417CrossRefGoogle Scholar
  6. 6.
    Marx J (2001) Caveolae: a once-elusive structure gets some respect. Science 294:1862–1865CrossRefGoogle Scholar
  7. 7.
    Sinha B, Koster D, Ruez R, Gonnord P, Bastiani M et al (2011) Cells respond to mechanical stress by rapid disassembly of caveolae. Cell 144:402–413CrossRefGoogle Scholar
  8. 8.
    Harvey RD, Calaghan SC (2012) Caveolae create local signalling domains through their distinct protein content, lipid profile and morphology. J Mol Cell Cardiol 52:366–375CrossRefGoogle Scholar
  9. 9.
    Schlegel A, Volonte D, Engelman JA, Galbiata F, Mehta P et al (1998) Crowded little caves: structure and function of caveolae. Cell Signal 10:457–463CrossRefGoogle Scholar
  10. 10.
    Anderson RG (1998) The caveolae membrane system. AnnuRevBiochem 67:199–225Google Scholar
  11. 11.
    Sengupta P, Philip F, Scarlata S (2008) Caveolin-1 alters Ca2+ signal duration through specific interaction with the G{alpha}q family of G proteins. J Cell Sci 121:1363–1372CrossRefGoogle Scholar
  12. 12.
    Ji G, Barsotti RJ, Feldman ME, Kotlikoff MI (2002) Stretch-induced calcium release in smooth muscle. J Gen Physiol 119:533–544CrossRefGoogle Scholar
  13. 13.
    Echarri A, Del Pozo MA (2015) Caveolae—mechanosensitive membrane invaginations linked to actin filaments. J Cell Sci 128:2747–2758CrossRefGoogle Scholar
  14. 14.
    Yang L, Scarlata S (2017) Super-resolution visualization of caveola deformation in response to osmotic stress. J Biol Chem 292:3779–3788CrossRefGoogle Scholar
  15. 15.
    Woodman SE, Park DS, Cohen AW, Cheung MW, Chandra M et al (2002) Caveolin-3 knock-out mice develop a progressive cardiomyopathy and show hyperactivation of the p42/44 MAPK cascade. J Biol Chem 277:38988–38997CrossRefGoogle Scholar
  16. 16.
    Dorn Ii GW, Brown JH (1999) Gq signaling in cardiac adaptation and maladaptation. Trends Cardiovasc Med 9:26–34CrossRefGoogle Scholar
  17. 17.
    Cirka H, Monterosso M, Diamantides N, Favreau J, Wen Q et al (2016) Active traction force response to long-term cyclic stretch is dependent on cell pre-stress. Biophys J 110:1845–1857CrossRefGoogle Scholar
  18. 18.
    Guo Y, Yang L, Haught K, Scarlata S (2015) Osmotic stress reduces Ca2+ signals through deformation of caveolae. J Biol Chem 290:16698–16707CrossRefGoogle Scholar
  19. 19.
    Volonte D, Galbiati F, Lisanti MP (1999) Visualization of caveolin-1, a caveolar marker protein, in living cells using green fluorescent protein (GFP) chimeras. The subcellular distribution of caveolin-1 is modulated by cell-cell contact. FEBS Lett 445:431–439CrossRefGoogle Scholar
  20. 20.
    Tang Z, Scherer PE, Okamoto T, Song K, Chu C et al (1996) Molecular cloning of caveolin-3, a novel member of the caveolin gene family expressed predominantly in muscle. J Biol Chem 271:2255–2261CrossRefGoogle Scholar
  21. 21.
    Gilbert G, Ducret T, Savineau JP, Marthan R, Quignard JF (2016) Caveolae are involved in mechanotransduction during pulmonary hypertension. Am J Physiol Lung Cell Mol Physiol 310:L1078–L1087CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Dept. of Chemistry and BiochemistryWorcester Polytechnic InstituteWorcesterUSA

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