Tissue inducible Lifeact expression allows visualization of actin dynamics in vivo and ex vivo
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 u...
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| Published in: | European journal of cell biology Vol. 91; no. 11-12; pp. 923 - 929 |
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| Main Authors: | , , , , , , , , , |
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| Language: | English |
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01-11-2012
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| 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. |
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| AbstractList | 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. |
| Author | Wedlich-Soldner, Roland Schachtner, Hannah Watson, Steve P. Stevenson, David Strathdee, Douglas Machesky, Laura M. Li, Ang Sixt, Michael Calaminus, Simon D.J. Thomas, Steven G. |
| AuthorAffiliation | 2 Centre for Cardiovascular Sciences, Institute of Biomedical Research, College of Medical and Dental Sciences, University of Birmingham, Birmingham, B15 2TT 3 Institute of Science and Technology, Am Campus 1, A-3400 Klosterneuberg, Austria 4 Cellular Dynamics and Cell Patterning, Max-Planck Institute of Biochemistry, Am, Klopferspitz 18, 82152 Martinsried, Germany 1 The Beatson Institute for Cancer Research, Garscube Estate, Switchback Road, Bearsden, Glasgow, G61 1BD |
| AuthorAffiliation_xml | – name: 3 Institute of Science and Technology, Am Campus 1, A-3400 Klosterneuberg, Austria – name: 4 Cellular Dynamics and Cell Patterning, Max-Planck Institute of Biochemistry, Am, Klopferspitz 18, 82152 Martinsried, Germany – name: 2 Centre for Cardiovascular Sciences, Institute of Biomedical Research, College of Medical and Dental Sciences, University of Birmingham, Birmingham, B15 2TT – name: 1 The Beatson Institute for Cancer Research, Garscube Estate, Switchback Road, Bearsden, Glasgow, G61 1BD |
| Author_xml | – sequence: 1 givenname: Hannah surname: Schachtner fullname: Schachtner, Hannah organization: The Beatson Institute for Cancer Research, Garscube Estate, Switchback Road, Bearsden, Glasgow, G61 1BD, United Kingdom – sequence: 2 givenname: Ang surname: Li fullname: Li, Ang organization: The Beatson Institute for Cancer Research, Garscube Estate, Switchback Road, Bearsden, Glasgow, G61 1BD, United Kingdom – sequence: 3 givenname: David surname: Stevenson fullname: Stevenson, David organization: The Beatson Institute for Cancer Research, Garscube Estate, Switchback Road, Bearsden, Glasgow, G61 1BD, United Kingdom – sequence: 4 givenname: Simon D.J. surname: Calaminus fullname: Calaminus, Simon D.J. organization: The Beatson Institute for Cancer Research, Garscube Estate, Switchback Road, Bearsden, Glasgow, G61 1BD, United Kingdom – sequence: 5 givenname: Steven G. surname: Thomas fullname: Thomas, Steven G. organization: Centre for Cardiovascular Sciences, Institute of Biomedical Research, College of Medical and Dental Sciences, University of Birmingham, B15 2TT, United Kingdom – sequence: 6 givenname: Steve P. surname: Watson fullname: Watson, Steve P. organization: Centre for Cardiovascular Sciences, Institute of Biomedical Research, College of Medical and Dental Sciences, University of Birmingham, B15 2TT, United Kingdom – sequence: 7 givenname: Michael surname: Sixt fullname: Sixt, Michael organization: Institute of Science and Technology, Am Campus 1, A-3400 Klosterneuberg, Austria – sequence: 8 givenname: Roland surname: Wedlich-Soldner fullname: Wedlich-Soldner, Roland organization: Cellular Dynamics and Cell Patterning, Max-Planck Institute of Biochemistry, Am Klopferspitz 18, 82152 Martinsried, Germany – sequence: 9 givenname: Douglas surname: Strathdee fullname: Strathdee, Douglas organization: The Beatson Institute for Cancer Research, Garscube Estate, Switchback Road, Bearsden, Glasgow, G61 1BD, United Kingdom – sequence: 10 givenname: Laura M. surname: Machesky fullname: Machesky, Laura M. email: l.machesky@beatson.gla.ac.uk organization: The Beatson Institute for Cancer Research, Garscube Estate, Switchback Road, Bearsden, Glasgow, G61 1BD, United Kingdom |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/22658956$$D View this record in MEDLINE/PubMed |
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| Keywords | Cytoskeleton Motility Cytokinesis Actin dynamics Imaging Cell migration |
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| SubjectTerms | Actin Cytoskeleton - ultrastructure Actin dynamics Actins - genetics Actins - metabolism Animals Cell Line Cell migration Cell Movement Cytokinesis Cytoskeleton Gene Expression Regulation Genes, Reporter Green Fluorescent Proteins - genetics Imaging Melanocytes - metabolism Melanocytes - ultrastructure Mice Mice, Inbred C57BL Mice, Transgenic Microscopy, Fluorescence Motility Neural Stem Cells - metabolism Neural Stem Cells - ultrastructure Neuropeptides - genetics Neuropeptides - metabolism Organ Specificity Peptides - genetics Pseudopodia - metabolism rac GTP-Binding Proteins - genetics rac GTP-Binding Proteins - metabolism rac1 GTP-Binding Protein Recombinant Proteins - genetics Time-Lapse Imaging Transcription, Genetic |
| Title | Tissue inducible Lifeact expression allows visualization of actin dynamics in vivo and ex vivo |
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