Formaldehyde fixation is detrimental to actin cables in glucose-depleted S. cerevisiae cells
Authors:Pavla Vasicova1,#, Mark Rinnerthaler2, Danusa Haskova1, Lenka Novakova1, Ivana Malcova1, Michael Breitenbach2, Jiri Hasek1
doi: 10.15698/mic2016.05.499
Volume 3, pp. 206 to 214, published 12/04/2016.
1 Laboratory of Cell Reproduction, Institute of Microbiology of the CAS, v.v.i., Prague, Czech Republic.
2 Department of Cell Biology, Division of Genetics, University of Salzburg, Salzburg, Austria.
# present address: Institute of Molecular Genetics of the CAS, v.v.i.
Keywords:
Abp140-GFP, Abp1-RFP, Actin cables, Actin patches, yeast.
Corresponding Author(s):
Conflict of interest statement:
The authors declare no conflict of interest.
Please cite this article as:
Pavla Vasicova, Mark Rinnerthaler, Danusa Haskova, Lenka Novakova, Ivana Malcova, Michael Breitenbach, Jiri Hasek (2016). Formaldehyde fixation is detrimental to actin cables in glucose-depleted S. cerevisiae cells. Microbial Cell 3(5): 206-214. doi: 10.15698/mic2016.05.499
© 2016 Vasicova et al. This is an open-access article released under the terms of the Creative Commons Attribution (CC BY) license, which allows the unrestricted use, distribution, and reproduction in any medium, provided the original author and source are acknowledged.
Abstract:
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.