Literature DB >> 22658956

Tissue inducible Lifeact expression allows visualization of actin dynamics in vivo and ex vivo.

Hannah Schachtner1, Ang Li1, David Stevenson1, Simon D J Calaminus1, Steve Thomas2, Steve P Watson2, Michael Sixt3, Roland Wedlich-Soldner4, Douglas Strathdee1, Laura M Machesky1.   

Abstract

We describe here the development and characterization of a conditionally inducible mouse model expressing Lifeact-GFP, a peptide that reports the dynamics of filamentous actin. We have used this model to study platelets, megakaryocytes and melanoblasts and we provide evidence that Lifeact-GFP is a useful reporter in these cell types ex vivo. In the case of platelets and megakaryocytes, these cells are not transfectable by traditional methods, so conditional activation of Lifeact allows the study of actin dynamics in these cells live. We studied melanoblasts in native skin explants from embryos, allowing the visualization of live actin dynamics during cytokinesis and migration. Our study revealed that melanoblasts lacking the small GTPase Rac1 show a delay in the formation of new pseudopodia following cytokinesis that accounts for the previously reported cytokinesis delay in these cells. Thus, through use of this mouse model, we were able to gain insights into the actin dynamics of cells that could only previously be studied using fixed specimens or following isolation from their native tissue environment.
Copyright © 2012. Published by Elsevier GmbH.

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Year:  2012        PMID: 22658956      PMCID: PMC3930012          DOI: 10.1016/j.ejcb.2012.04.002

Source DB:  PubMed          Journal:  Eur J Cell Biol        ISSN: 0171-9335            Impact factor:   4.492


  37 in total

1.  Microfilament dynamics during cell movement and chemotaxis monitored using a GFP-actin fusion protein.

Authors:  M Westphal; A Jungbluth; M Heidecker; B Mühlbauer; C Heizer; J M Schwartz; G Marriott; G Gerisch
Journal:  Curr Biol       Date:  1997-03-01       Impact factor: 10.834

2.  Identification of a novel, actin-rich structure, the actin nodule, in the early stages of platelet spreading.

Authors:  S D J Calaminus; S Thomas; O J T McCarty; L M Machesky; S P Watson
Journal:  J Thromb Haemost       Date:  2008-08-28       Impact factor: 5.824

3.  Targeting of human eNOS promoter to the Hprt locus of mice leads to tissue-restricted transgene expression.

Authors:  P V Guillot; L Liu; J A Kuivenhoven; J Guan; R D Rosenberg; W C Aird
Journal:  Physiol Genomics       Date:  2000-03-13       Impact factor: 3.107

4.  Spontaneous malignant transformation of melanocytes explanted from Wf/Wf mice with a Kit kinase-domain mutation.

Authors:  L Larue; N Dougherty; S Porter; B Mintz
Journal:  Proc Natl Acad Sci U S A       Date:  1992-08-15       Impact factor: 11.205

5.  Cofilin is a component of intranuclear and cytoplasmic actin rods induced in cultured cells.

Authors:  E Nishida; K Iida; N Yonezawa; S Koyasu; I Yahara; H Sakai
Journal:  Proc Natl Acad Sci U S A       Date:  1987-08       Impact factor: 11.205

6.  Analysis of human flap endonuclease 1 mutants reveals a mechanism to prevent triplet repeat expansion.

Authors:  Yuan Liu; Robert A Bambara
Journal:  J Biol Chem       Date:  2003-01-28       Impact factor: 5.157

7.  Fluorescence microscopic localization of actin in pollen tubes: comparison of actin antibody and phalloidin staining.

Authors:  X J Tang; S A Lancelle; P K Hepler
Journal:  Cell Motil Cytoskeleton       Date:  1989

8.  Actin dynamics in living mammalian cells.

Authors:  C Ballestrem; B Wehrle-Haller; B A Imhof
Journal:  J Cell Sci       Date:  1998-06       Impact factor: 5.285

9.  Actin filament severing by cofilin is more important for assembly than constriction of the cytokinetic contractile ring.

Authors:  Qian Chen; Thomas D Pollard
Journal:  J Cell Biol       Date:  2011-10-24       Impact factor: 10.539

10.  Application of Lifeact reveals F-actin dynamics in Arabidopsis thaliana and the liverwort, Marchantia polymorpha.

Authors:  Atsuko Era; Motoki Tominaga; Kazuo Ebine; Chie Awai; Chieko Saito; Kimitsune Ishizaki; Katsuyuki T Yamato; Takayuki Kohchi; Akihiko Nakano; Takashi Ueda
Journal:  Plant Cell Physiol       Date:  2009-04-15       Impact factor: 4.927

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  16 in total

Review 1.  Munc18c: a controversial regulator of peripheral insulin action.

Authors:  Latha Ramalingam; Stephanie M Yoder; Eunjin Oh; Debbie C Thurmond
Journal:  Trends Endocrinol Metab       Date:  2014-07-12       Impact factor: 12.015

Review 2.  Getting a good view: in vitro imaging of platelets under flow.

Authors:  Oluwamayokun Oshinowo; Tamara Lambert; Yumiko Sakurai; Renee Copeland; Caroline E Hansen; Wilbur A Lam; David R Myers
Journal:  Platelets       Date:  2020-02-28       Impact factor: 3.862

3.  Intravital Imaging of Bone Marrow Niches.

Authors:  Myriam L R Haltalli; Cristina Lo Celso
Journal:  Methods Mol Biol       Date:  2021

4.  Intravital Microscopy for Hematopoietic Studies.

Authors:  Myriam L R Haltalli; Cristina Lo Celso
Journal:  Methods Mol Biol       Date:  2023

5.  Ex vivo culture of mouse embryonic skin and live-imaging of melanoblast migration.

Authors:  Richard L Mort; Margaret Keighren; Leonard Hay; Ian J Jackson
Journal:  J Vis Exp       Date:  2014-05-19       Impact factor: 1.355

6.  Mouse Tafazzin Is Required for Male Germ Cell Meiosis and Spermatogenesis.

Authors:  Laurence C Cadalbert; Farah Naz Ghaffar; David Stevenson; Sheila Bryson; Frédéric M Vaz; Eyal Gottlieb; Douglas Strathdee
Journal:  PLoS One       Date:  2015-06-26       Impact factor: 3.240

7.  Accessible Synthetic Probes for Staining Actin inside Platelets and Megakaryocytes by Employing Lifeact Peptide.

Authors:  Lucia Cardo; Steve G Thomas; Alexandra Mazharian; Zoe Pikramenou; Joshua Z Rappoport; Michael J Hannon; Stephen P Watson
Journal:  Chembiochem       Date:  2015-06-15       Impact factor: 3.164

8.  Platelet actin nodules are podosome-like structures dependent on Wiskott-Aldrich syndrome protein and ARP2/3 complex.

Authors:  Natalie S Poulter; Alice Y Pollitt; Amy Davies; Dessislava Malinova; Gerard B Nash; Mike J Hannon; Zoe Pikramenou; Joshua Z Rappoport; John H Hartwig; Dylan M Owen; Adrian J Thrasher; Stephen P Watson; Steven G Thomas
Journal:  Nat Commun       Date:  2015-06-01       Impact factor: 14.919

9.  In vivo imaging of Nematostella vectensis embryogenesis and late development using fluorescent probes.

Authors:  Timothy Q DuBuc; Ada A Dattoli; Leslie S Babonis; Miguel Salinas-Saavedra; Eric Röttinger; Mark Q Martindale; Marten Postma
Journal:  BMC Cell Biol       Date:  2014-11-30       Impact factor: 4.241

10.  Pdlim7 Regulates Arf6-Dependent Actin Dynamics and Is Required for Platelet-Mediated Thrombosis in Mice.

Authors:  Alexander E Urban; Erin O Quick; Kaylie P Miller; Jennifer Krcmery; Hans-Georg Simon
Journal:  PLoS One       Date:  2016-10-28       Impact factor: 3.240

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