Formaldehyde fixation is detrimental to actin cables in glucose-depleted S. cerevisiae cells
Actin filaments form cortical patches and emanating cables in fermenting cells of Saccharomyces cerevisiae. This pattern has been shown to be depolarized in glucose-depleted cells after formaldehyde fixation and staining with rhodamine-tagged phalloidin. Loss of actin cables in mother cells was rema...
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Shared Science Publishers OG
2016-04-01
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| Series: | Microbial Cell |
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| Online Access: | http://microbialcell.com/researcharticles/formaldehyde-fixation-is-detrimental-to-actin-cables-in-glucose-depleted-s-cerevisiae-cells/ |
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| author | Pavla Vasicova Mark Rinnerthaler Danusa Haskova Lenka Novakova Ivana Malcova Michael Breitenbach Jiri Hasek |
| author_facet | Pavla Vasicova Mark Rinnerthaler Danusa Haskova Lenka Novakova Ivana Malcova Michael Breitenbach Jiri Hasek |
| author_sort | Pavla Vasicova |
| collection | DOAJ |
| description | Actin filaments form cortical patches and emanating cables in fermenting cells of Saccharomyces cerevisiae. This pattern has been shown to be depolarized in glucose-depleted cells after formaldehyde fixation and staining with rhodamine-tagged phalloidin. Loss of actin cables in mother cells was remarkable. Here we extend our knowledge on actin in live glucose-depleted cells co-expressing the marker of actin patches (Abp1-RFP) with the marker of actin cables (Abp140-GFP). Glucose depletion resulted in appearance of actin patches also in mother cells. However, even after 80 min of glucose deprivation these cells showed a clear network of actin cables labeled with Abp140-GFP in contrast to previously published data. In live cells with a mitochondrial dysfunction (rho0 cells), glucose depletion resulted in almost immediate appearance of Abp140-GFP foci partially overlapping with Abp1-RFP patches in mother cells. Residual actin cables were clustered in patch-associated bundles. A similar overlapping “patchy” pattern of both actin markers was observed upon treatment of glucose-deprived rho+ cells with FCCP (the inhibitor of oxidative phosphorylation) and upon treatment with formaldehyde. While the formaldehyde-targeted process stays unknown, our results indicate that published data on yeast actin cytoskeleton obtained from glucose-depleted cells after fixation should be considered with caution. |
| format | Article |
| id | doaj-art-debee9fd75584d55a0bb7ecdcc1c4c51 |
| institution | OA Journals |
| issn | 2311-2638 |
| language | English |
| publishDate | 2016-04-01 |
| publisher | Shared Science Publishers OG |
| record_format | Article |
| series | Microbial Cell |
| spelling | doaj-art-debee9fd75584d55a0bb7ecdcc1c4c512025-08-20T02:04:34ZengShared Science Publishers OGMicrobial Cell2311-26382016-04-013520621410.15698/mic2016.05.499Formaldehyde fixation is detrimental to actin cables in glucose-depleted S. cerevisiae cellsPavla Vasicova0Mark Rinnerthaler1Danusa Haskova2Lenka Novakova3Ivana Malcova4Michael Breitenbach5Jiri Hasek6Laboratory of Cell Reproduction, Institute of Microbiology of the CAS, v.v.i., Prague, Czech Republic.Department of Cell Biology, Division of Genetics, University of Salzburg, Salzburg, Austria.Laboratory of Cell Reproduction, Institute of Microbiology of the CAS, v.v.i., Prague, Czech Republic.Laboratory of Cell Reproduction, Institute of Microbiology of the CAS, v.v.i., Prague, Czech Republic.Laboratory of Cell Reproduction, Institute of Microbiology of the CAS, v.v.i., Prague, Czech Republic.Department of Cell Biology, Division of Genetics, University of Salzburg, Salzburg, Austria.Laboratory of Cell Reproduction, Institute of Microbiology of the CAS, v.v.i., Prague, Czech Republic.Actin filaments form cortical patches and emanating cables in fermenting cells of Saccharomyces cerevisiae. This pattern has been shown to be depolarized in glucose-depleted cells after formaldehyde fixation and staining with rhodamine-tagged phalloidin. Loss of actin cables in mother cells was remarkable. Here we extend our knowledge on actin in live glucose-depleted cells co-expressing the marker of actin patches (Abp1-RFP) with the marker of actin cables (Abp140-GFP). Glucose depletion resulted in appearance of actin patches also in mother cells. However, even after 80 min of glucose deprivation these cells showed a clear network of actin cables labeled with Abp140-GFP in contrast to previously published data. In live cells with a mitochondrial dysfunction (rho0 cells), glucose depletion resulted in almost immediate appearance of Abp140-GFP foci partially overlapping with Abp1-RFP patches in mother cells. Residual actin cables were clustered in patch-associated bundles. A similar overlapping “patchy” pattern of both actin markers was observed upon treatment of glucose-deprived rho+ cells with FCCP (the inhibitor of oxidative phosphorylation) and upon treatment with formaldehyde. While the formaldehyde-targeted process stays unknown, our results indicate that published data on yeast actin cytoskeleton obtained from glucose-depleted cells after fixation should be considered with caution.http://microbialcell.com/researcharticles/formaldehyde-fixation-is-detrimental-to-actin-cables-in-glucose-depleted-s-cerevisiae-cells/Abp140-GFPAbp1-RFPActin cables Actin patchesyeast |
| spellingShingle | Pavla Vasicova Mark Rinnerthaler Danusa Haskova Lenka Novakova Ivana Malcova Michael Breitenbach Jiri Hasek Formaldehyde fixation is detrimental to actin cables in glucose-depleted S. cerevisiae cells Microbial Cell Abp140-GFP Abp1-RFP Actin cables Actin patches yeast |
| title | Formaldehyde fixation is detrimental to actin cables in glucose-depleted S. cerevisiae cells |
| title_full | Formaldehyde fixation is detrimental to actin cables in glucose-depleted S. cerevisiae cells |
| title_fullStr | Formaldehyde fixation is detrimental to actin cables in glucose-depleted S. cerevisiae cells |
| title_full_unstemmed | Formaldehyde fixation is detrimental to actin cables in glucose-depleted S. cerevisiae cells |
| title_short | Formaldehyde fixation is detrimental to actin cables in glucose-depleted S. cerevisiae cells |
| title_sort | formaldehyde fixation is detrimental to actin cables in glucose depleted s cerevisiae cells |
| topic | Abp140-GFP Abp1-RFP Actin cables Actin patches yeast |
| url | http://microbialcell.com/researcharticles/formaldehyde-fixation-is-detrimental-to-actin-cables-in-glucose-depleted-s-cerevisiae-cells/ |
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