Differential effects of LifeAct-GFP and actin-GFP on cell mechanics assessed using micropipette aspiration

Journal of Biomechanics
Kristina SliogeryteMartin M Knight

Abstract

The actin cytoskeleton forms a dynamic structure involved in many fundamental cellular processes including the control of cell morphology, migration and biomechanics. Recently LifeAct-GFP (green fluorescent protein) has been proposed for visualising actin structure and dynamics in live cells as an alternative to actin-GFP which has been shown to affect cell mechanics. Here we compare the two approaches in terms of their effect on cellular mechanical behaviour. Human mesenchymal stem cells (hMSCs) were analysed using micropipette aspiration and the effective cellular equilibrium and instantaneous moduli calculated using the standard linear solid model. We show that LifeAct-GFP provides clearer visualisation of F-actin organisation and dynamics. Furthermore, LifeAct-GFP does not alter effective cellular mechanical properties whereas actin-GFP expression causes an increase in the cell modulus. Interestingly, LifeAct-GFP expression did produce a small (~10%) increase in the percentage of cells exhibiting aspiration-induced membrane bleb formation, whilst actin-GFP expression reduced blebbing. Further studies examined the influence of LifeAct-GFP in other cell types, namely chondrogenically differentiated hMSCs and murine chondrocyt...Continue Reading

References

Aug 1, 1994·Biophysical Journal·D V ZhelevR M Hochmuth
Feb 24, 2001·Journal of Orthopaedic Research : Official Publication of the Orthopaedic Research Society·W R TrickeyF Guilak
Jul 9, 2004·Trends in Biochemical Sciences·Paul A Janmey, David A Weitz
Feb 16, 2006·The International Journal of Developmental Biology·Donald E Ingber
Apr 7, 2007·ASAIO Journal : a Peer-reviewed Journal of the American Society for Artificial Internal Organs·Gregory YourekJeremy J Mao
Aug 10, 2007·Cell Motility and the Cytoskeleton·Brian M BurkelWilliam M Bement
Jun 10, 2008·Nature Methods·Julia RiedlRoland Wedlich-Soldner
Jun 11, 2008·The Journal of Cell Biology·Oliver T Fackler, Robert Grosse
Jul 23, 2008·BMC Cell Biology·Samuel C W TanKin Liao
May 20, 2009·Journal of Biomechanical Engineering·Gidon OfekKyriacos A Athanasiou
Dec 8, 2009·Science·Thomas D Pollard, John A Cooper
Feb 2, 2010·Biochemical and Biophysical Research Communications·Haiyang YuLay Poh Tan
Mar 3, 2010·Nature Methods·Julia RiedlRoland Wedlich-Söldner
Aug 18, 2010·Proceedings of the National Academy of Sciences of the United States of America·Jan BruguésPierre Sens
Dec 24, 2010·Physical Biology·Hanna EngelkeJoachim O Rädler
Jan 10, 2012·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Angus K T WannMartin M Knight
Jun 12, 2013·Current Biology : CB·Chi Wai LeeJames Q Zheng
Dec 19, 2013·Communicative & Integrative Biology·Kristina Haase, Andrew E Pelling
May 27, 2014·Nature Methods·Gražvydas LukinavičiusKai Johnsson
Sep 1, 2015·Bioarchitecture·Brittany J BelinR Dyche Mullins

❮ Previous
Next ❯

Citations

Sep 16, 2018·Nature Protocols·Yuri M EfremovArvind Raman
Mar 29, 2019·Proceedings of the Institution of Mechanical Engineers. Part H, Journal of Engineering in Medicine·Ashkan HeydarianBehnoosh Vasaghi-Gharamaleki
Jul 13, 2019·Integrative Biology : Quantitative Biosciences From Nano to Macro·Andreas MüllerTilo Pompe
Sep 3, 2020·International Journal of Molecular Sciences·Haibo HuangLining Sun
Mar 3, 2019·Scientific Reports·Luis R FloresNúria Gavara
Apr 10, 2019·Scientific Reports·Yuri M EfremovArvind Raman
Jan 14, 2017·Journal of Cell Science·Michael MelakRobert Grosse
Nov 21, 2020·PLoS Biology·Alexander BelyyStefan Raunser
Oct 21, 2020·Biotechnology Advances·Yansheng HaoMing Li

❮ Previous
Next ❯

Methods Mentioned

BETA
transfection
super-resolution microscopy
confocal microscopy

Software Mentioned

GraphPad Prism
Matlab
LifeAct

Related Concepts

Related Feeds

Biomechanics

Biomechanics examines the generation of internal forces within the body and investigates the effects and control of forces that act on or are produced on tissues. Here are the latest discoveries.

Cell Migration

Cell migration is involved in a variety of physiological and pathological processes such as embryonic development, cancer metastasis, blood vessel formation and remoulding, tissue regeneration, immune surveillance and inflammation. Here is the latest research.

Related Papers

Conference Proceedings : ... Annual International Conference of the IEEE Engineering in Medicine and Biology Society
Wenxiao PanKam Leong
© 2022 Meta ULC. All rights reserved