A red fluorescent lifeact marker to study actin morphology in podocytes
Abstract F-actin is a major component of the cellular cytoskeleton, responsible for maintaining cell shape, enabling movement and facilitating intracellular transport. In the kidney, glomerular podocytes are highly dependent on their actin cytoskeleton shaping their unique foot processes. Hereditary...
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Nature Portfolio
2025-04-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-025-96822-w |
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| author | Eva Wiesner Julia Binz-Lotter Simon E. Tröder David Unnersjö-Jess Nelli Rutkowski Branko Zevnik Bernhard Schermer Thomas Benzing Roland Wedlich-Söldner Matthias J. Hackl |
| author_facet | Eva Wiesner Julia Binz-Lotter Simon E. Tröder David Unnersjö-Jess Nelli Rutkowski Branko Zevnik Bernhard Schermer Thomas Benzing Roland Wedlich-Söldner Matthias J. Hackl |
| author_sort | Eva Wiesner |
| collection | DOAJ |
| description | Abstract F-actin is a major component of the cellular cytoskeleton, responsible for maintaining cell shape, enabling movement and facilitating intracellular transport. In the kidney, glomerular podocytes are highly dependent on their actin cytoskeleton shaping their unique foot processes. Hereditary mutations in actin-binding proteins cause focal segmental glomerulosclerosis, while other organs remain largely unaffected. So far, actin visualization in podocytes has been limited to electron microscopy or indirect immunofluorescent labeling of actin-binding proteins. However, the short F-actin-binding peptide Lifeact enables researchers to study actin dynamics in vitro and in vivo with minimal interference with actin metabolism. Here we introduce a new mouse model with conditional expression of a Lifeact.mScarlet-I fusion protein providing red labeling of actin. Cre recombinase-mediated activity allows cell-specific and mosaic expression in podocytes, enabling selective labeling of individual cells to contrast with non-expressing neighboring cells. Transgenic mice are born healthy and young animals display no kidney-related phenotype. By intravital imaging and super-resolution microscopy, we show subcellular localization of actin to the foot processes in a resolution previously only obtainable by electron microscopy. Our novel mouse line provides the opportunity to study the actin cytoskeleton in podocytes and other cell types by intravital imaging and other conventional light microscopy techniques. |
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| id | doaj-art-28f14d76426748d48c2ef9db583d913e |
| institution | OA Journals |
| issn | 2045-2322 |
| language | English |
| publishDate | 2025-04-01 |
| publisher | Nature Portfolio |
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| spelling | doaj-art-28f14d76426748d48c2ef9db583d913e2025-08-20T02:12:01ZengNature PortfolioScientific Reports2045-23222025-04-0115111310.1038/s41598-025-96822-wA red fluorescent lifeact marker to study actin morphology in podocytesEva Wiesner0Julia Binz-Lotter1Simon E. Tröder2David Unnersjö-Jess3Nelli Rutkowski4Branko Zevnik5Bernhard Schermer6Thomas Benzing7Roland Wedlich-Söldner8Matthias J. Hackl9Department II of Internal Medicine and Center for Molecular Medicine Cologne, Faculty of Medicine and University Hospital Cologne, University of CologneDepartment II of Internal Medicine and Center for Molecular Medicine Cologne, Faculty of Medicine and University Hospital Cologne, University of CologneCluster of Excellence Cellular Stress Responses in Aging-Associated Diseases (CECAD), University of CologneDepartment II of Internal Medicine and Center for Molecular Medicine Cologne, Faculty of Medicine and University Hospital Cologne, University of CologneDepartment II of Internal Medicine and Center for Molecular Medicine Cologne, Faculty of Medicine and University Hospital Cologne, University of CologneCluster of Excellence Cellular Stress Responses in Aging-Associated Diseases (CECAD), University of CologneDepartment II of Internal Medicine and Center for Molecular Medicine Cologne, Faculty of Medicine and University Hospital Cologne, University of CologneDepartment II of Internal Medicine and Center for Molecular Medicine Cologne, Faculty of Medicine and University Hospital Cologne, University of CologneInstitute of Cell Dynamics and Imaging and Cells in Motion Interfaculty Centre, University of MünsterDepartment II of Internal Medicine and Center for Molecular Medicine Cologne, Faculty of Medicine and University Hospital Cologne, University of CologneAbstract F-actin is a major component of the cellular cytoskeleton, responsible for maintaining cell shape, enabling movement and facilitating intracellular transport. In the kidney, glomerular podocytes are highly dependent on their actin cytoskeleton shaping their unique foot processes. Hereditary mutations in actin-binding proteins cause focal segmental glomerulosclerosis, while other organs remain largely unaffected. So far, actin visualization in podocytes has been limited to electron microscopy or indirect immunofluorescent labeling of actin-binding proteins. However, the short F-actin-binding peptide Lifeact enables researchers to study actin dynamics in vitro and in vivo with minimal interference with actin metabolism. Here we introduce a new mouse model with conditional expression of a Lifeact.mScarlet-I fusion protein providing red labeling of actin. Cre recombinase-mediated activity allows cell-specific and mosaic expression in podocytes, enabling selective labeling of individual cells to contrast with non-expressing neighboring cells. Transgenic mice are born healthy and young animals display no kidney-related phenotype. By intravital imaging and super-resolution microscopy, we show subcellular localization of actin to the foot processes in a resolution previously only obtainable by electron microscopy. Our novel mouse line provides the opportunity to study the actin cytoskeleton in podocytes and other cell types by intravital imaging and other conventional light microscopy techniques.https://doi.org/10.1038/s41598-025-96822-w |
| spellingShingle | Eva Wiesner Julia Binz-Lotter Simon E. Tröder David Unnersjö-Jess Nelli Rutkowski Branko Zevnik Bernhard Schermer Thomas Benzing Roland Wedlich-Söldner Matthias J. Hackl A red fluorescent lifeact marker to study actin morphology in podocytes Scientific Reports |
| title | A red fluorescent lifeact marker to study actin morphology in podocytes |
| title_full | A red fluorescent lifeact marker to study actin morphology in podocytes |
| title_fullStr | A red fluorescent lifeact marker to study actin morphology in podocytes |
| title_full_unstemmed | A red fluorescent lifeact marker to study actin morphology in podocytes |
| title_short | A red fluorescent lifeact marker to study actin morphology in podocytes |
| title_sort | red fluorescent lifeact marker to study actin morphology in podocytes |
| url | https://doi.org/10.1038/s41598-025-96822-w |
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