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Three-dimensional and immune electron microscopic analysis of the secretory pathway in Saccharomyces cerevisiae | Histochemistry and Cell Biology
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Until now, the mechanisms of ER-to-Golgi and intra-Golgi transport remain obscure. This is especially evident for the Golgi of S. cerevisiae where different Golgi compartments are not organized in stacks. Here, using improved sample preparation protocols, we examined the 3D organization of pre-Golgi and Golgi compartments and found several new features of the structures functioning along the secretory pathway. In the cytoplasmic sheet ER, we found narrow pores that aggregated near the rims, and tubular networks tightly interconnected with sheets of several cytoplasmic ER cisternae. Within the Golgi compartments, we found disks with wide pores, disks with narrow pores, and disk-like networks with varicosities or nodules at the point of branching and thick membranes. Sometimes, these compartments contained 30 nm buds coated with a clathrin-like coat. The lumen of these Golgi compartments was more osmiophilic than the lumen of the ER. In contrast to ER elements, Golgi compartments were isolated and in the majority of cases not connected, although we observed some connections between Golgi compartments and also between Golgi disks with wide pores and the ER. Two types of free vesicles of 35–40 and 45–50 nm were found, the former being sometimes partially coated with a clathrin-like coat. Sec31, a COPII component, was found near narrow pores in the cytoplasmic sheets of the ER, over edge aggregates of narrow pores, and within the ER network. The cis-Golgi marker Rer1p was detected on disks or semi-spheres with wide pores, while the medial Golgi marker Gos1p was found on disks or semi-spheres with narrow pores. Gos1p was found to be enriched on 45–50 nm vesicles, while Rer1p was depleted. The 35–40 nm vesicles did not show either label. These findings are discussed from the point of view of mechanisms of transport.
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pubmed, article, google, scholar, cas, golgi, cell, mironov, biol, yeast, central, electron, vesicles, microscopy, beznoussenko, transport, cells, arrow, cerevisiae, compartments, luini, sections, saccharomyces, pores, structure, tomography, secretory, intragolgi, membrane, methods, polishchuk, traffic, fig, found, medial, access, endoplasmic, reticulum, sci, fusella, glick, panels, threedimensional, pathway, mol, giandomenico, apparatus, italy, scale, panel,
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month download article/chapter correlative video–light–electron microscopy related subjects double-step image acquisition inhibits intra–golgi transport time–resolved electron tomography microtubule–deprived living cells activated mono–adp–ribosylation high-pressure-freezing experiments full article pdf high–pressure frozen samples cis-golgi marker rer1p check access instant access intra–golgi transport privacy choices/manage cookies copi-coated bud 3d electron microscopy 3d protein localization antonella ragnini-wilson stimulated venus flytrap acknowledge italian firc mironov aa jr yeast endoplasmic reticulum er exit sites article histochemistry golgi structure correlates apical plasma membrane plasma membrane reshaping cellular electron tomography article beznoussenko intracellular transport theme scanning electron microscopy transmission electron microscopy routine electron microscopy cis-golgi contained intra-golgi traffic copi-dependent vesicle golgi maturation visualized medial golgi marker martínez-menárguez ja high-pressure freezing article log qb–snares gs27 cellulose capillary tubes ribosome–excluding scaffold day kj koster aj pug36-gos1 construct ifom-ieo campus
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- Hawes C (2012) The ER/Golgi interface–is there anything in-between?
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headline:Three-dimensional and immune electron microscopic analysis of the secretory pathway in Saccharomyces cerevisiae
description:Until now, the mechanisms of ER-to-Golgi and intra-Golgi transport remain obscure. This is especially evident for the Golgi of S. cerevisiae where different Golgi compartments are not organized in stacks. Here, using improved sample preparation protocols, we examined the 3D organization of pre-Golgi and Golgi compartments and found several new features of the structures functioning along the secretory pathway. In the cytoplasmic sheet ER, we found narrow pores that aggregated near the rims, and tubular networks tightly interconnected with sheets of several cytoplasmic ER cisternae. Within the Golgi compartments, we found disks with wide pores, disks with narrow pores, and disk-like networks with varicosities or nodules at the point of branching and thick membranes. Sometimes, these compartments contained 30 nm buds coated with a clathrin-like coat. The lumen of these Golgi compartments was more osmiophilic than the lumen of the ER. In contrast to ER elements, Golgi compartments were isolated and in the majority of cases not connected, although we observed some connections between Golgi compartments and also between Golgi disks with wide pores and the ER. Two types of free vesicles of 35–40 and 45–50 nm were found, the former being sometimes partially coated with a clathrin-like coat. Sec31, a COPII component, was found near narrow pores in the cytoplasmic sheets of the ER, over edge aggregates of narrow pores, and within the ER network. The cis-Golgi marker Rer1p was detected on disks or semi-spheres with wide pores, while the medial Golgi marker Gos1p was found on disks or semi-spheres with narrow pores. Gos1p was found to be enriched on 45–50 nm vesicles, while Rer1p was depleted. The 35–40 nm vesicles did not show either label. These findings are discussed from the point of view of mechanisms of transport.
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headline:Three-dimensional and immune electron microscopic analysis of the secretory pathway in Saccharomyces cerevisiae
description:Until now, the mechanisms of ER-to-Golgi and intra-Golgi transport remain obscure. This is especially evident for the Golgi of S. cerevisiae where different Golgi compartments are not organized in stacks. Here, using improved sample preparation protocols, we examined the 3D organization of pre-Golgi and Golgi compartments and found several new features of the structures functioning along the secretory pathway. In the cytoplasmic sheet ER, we found narrow pores that aggregated near the rims, and tubular networks tightly interconnected with sheets of several cytoplasmic ER cisternae. Within the Golgi compartments, we found disks with wide pores, disks with narrow pores, and disk-like networks with varicosities or nodules at the point of branching and thick membranes. Sometimes, these compartments contained 30 nm buds coated with a clathrin-like coat. The lumen of these Golgi compartments was more osmiophilic than the lumen of the ER. In contrast to ER elements, Golgi compartments were isolated and in the majority of cases not connected, although we observed some connections between Golgi compartments and also between Golgi disks with wide pores and the ER. Two types of free vesicles of 35–40 and 45–50 nm were found, the former being sometimes partially coated with a clathrin-like coat. Sec31, a COPII component, was found near narrow pores in the cytoplasmic sheets of the ER, over edge aggregates of narrow pores, and within the ER network. The cis-Golgi marker Rer1p was detected on disks or semi-spheres with wide pores, while the medial Golgi marker Gos1p was found on disks or semi-spheres with narrow pores. Gos1p was found to be enriched on 45–50 nm vesicles, while Rer1p was depleted. The 35–40 nm vesicles did not show either label. These findings are discussed from the point of view of mechanisms of transport.
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