ArticleThe journal of histochemistry and cytochemistry : official journal of the Histochemistry Society2009
Ultrastructural analysis of nanogold-labeled endocytic compartments of yeast Saccharomyces cerevisiae using a cryosectioning procedure.
Article in The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society, 2009. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 10 papers.
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10 citing papers in PubMed, 15 citations in OpenAlex.
- The GTPase activating protein Gyp7 regulates Rab7/Ypt7 activity on late endosomes.The Journal of cell biology · 2024Article
- The HOPS tethering complex is required to maintain signaling endosome identity and TORC1 activity.The Journal of cell biology · 2022Article
- Retromer-driven membrane tubulation separates endosomal recycling from Rab7/Ypt7-dependent fusion.Molecular biology of the cell · 2017Article
- Spatiotemporal dynamics of membrane remodeling and fusion proteins during endocytic transport.Molecular biology of the cell · 2015Article
- Nanogold labeling of the yeast endosomal system for ultrastructural analyses.Journal of visualized experiments : JoVE · 2014Article
- Identification of gold nanoparticle-resistant mutants of Saccharomyces cerevisiae suggests a role for respiratory metabolism in mediating toxicity.Applied and environmental microbiology · 2013Article
- A dual role for K63-linked ubiquitin chains in multivesicular body biogenesis and cargo sorting.Molecular biology of the cell · 2012Article
- The role of cell wall revealed by the visualization of Saccharomyces cerevisiae transformation.Current microbiology · 2011Article
- Phosphorylation of a membrane curvature-sensing motif switches function of the HOPS subunit Vps41 in membrane tethering.The Journal of cell biology · 2010Article
- The CORVET subunit Vps8 cooperates with the Rab5 homolog Vps21 to induce clustering of late endosomal compartments.Molecular biology of the cell · 2009Article
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2 authors at 1 institution in 1 country.
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Abstract
Yeast Saccharomyces cerevisiae has been a valuable model organism for the study of the endosomal system of eukaryotic cells. Morphological analyses, however, have been limited because of the lack of specific protein markers and of procedures that lead to a satisfactory ultrastructural resolution. We have recently developed an immunoelectron microscopy (IEM) protocol adapted from the Tokuyasu method to prepare cryosections from mildly fixed yeast. This novel approach allows excellent cell preservation and a unique resolution of the yeast morphology. Here, we present a protocol that combines this procedure with the specific labeling of the various endosomal compartments with positively charged Nanogold. In particular, we show that this new protocol generates excellent results when applied for the examination of early and late endosomes, and of mutants with an endosomal trafficking defect. Importantly, this method is compatible with immunogold labeling of protein markers, and it is consequently appropriate for localization studies of both resident and cargo proteins. This new IEM protocol will be a valuable tool for the large community of scientists using yeast as a model system to investigate the membrane transport and the biogenesis of the endosomal system.
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