title: Formaldehyde fixation is detrimental to actin cables in glucose-depletedS. cerevisiaecells.
abstract: Actin filaments form cortical patches and emanating cables in fermenting cells ofSaccharomyces 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 (rho0cells), 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.
authors.affiliation: Laboratory of Cell Reproduction, Institute of Microbiology of the CAS, v.v.i., Prague, Czech Republic.
authors.forename: Pavla
authors.initials: P
authors.lastname: Vasicova
authors.affiliation: Department of Cell Biology, Division of Genetics, University of Salzburg, Salzburg, Austria.
authors.forename: Mark
authors.initials: M
authors.lastname: Rinnerthaler
authors.affiliation: Laboratory of Cell Reproduction, Institute of Microbiology of the CAS, v.v.i., Prague, Czech Republic.
authors.forename: Danusa
authors.initials: D
authors.lastname: Haskova
authors.affiliation: Laboratory of Cell Reproduction, Institute of Microbiology of the CAS, v.v.i., Prague, Czech Republic.
authors.forename: Lenka
authors.initials: L
authors.lastname: Novakova
authors.affiliation: Laboratory of Cell Reproduction, Institute of Microbiology of the CAS, v.v.i., Prague, Czech Republic.
authors.forename: Ivana
authors.initials: I
authors.lastname: Malcova
authors.affiliation: Department of Cell Biology, Division of Genetics, University of Salzburg, Salzburg, Austria.
authors.forename: Michael
authors.initials: M
authors.lastname: Breitenbach
authors.affiliation: Laboratory of Cell Reproduction, Institute of Microbiology of the CAS, v.v.i., Prague, Czech Republic.
authors.forename: Jiri
authors.initials: J
authors.lastname: Hasek
keywords: Abp1-RFP; Abp140-GFP; Actin cables; Actin patches; yeast