Compartmentalization of intracellular membrane glycocomponents is revealed by fracture-label.

Torrisi, Maria Rosaria; Silva, P Pinto da

doi:10.1083/jcb.98.1.29

Cited by 28 publications

(14 citation statements)

References 33 publications

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“…The intracellular membranes of various secretory and nonsecretory cells can be divided in two compartments: One defined by the absence of WGA binding sites, and the other by their presence. The first compartment comprises nuclear envelopes, endoplasmic reticulum membranes, mitochondria, and peroxisomes; the second includes the membranes of lysosomes, secretory granules, and small vesicles of the Golgi areas (Pinto da Silva and Torrisi and Pinto da Silva, 1984). The WGA fracture-labeled intracellular membranes of lymphocytes confirm the existence of the two compartments.…”

Section: Labelingmentioning

confidence: 89%

See 1 more Smart Citation

Wheat germ agglutinin fracture‐label of lymphocyte plasma and intracellular membranes

Torrisi¹,

Pavan²,

Lotti³

et al. 1986

J. Elec. Microsc. Tech.

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Conventional lectin cytochemistry or immunocytochemistry can be associated with freeze-fracture in fracture-label techniques: The thin section fracture-label and the critical point drying fracture-label. In the first method, which is particularly useful for investigating intracellular membranes, cells or tissues are freeze-fractured, thawed, labeled, and embedded in resin. Wheat germ agglutinin labeling on freeze-fractured human lymphocytes confirms the existence of two compartments of intracellular membranes, based on the sialoglycocomponent content.

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Section: Labelingmentioning

confidence: 89%

“…The results confirm the compartmentalization of membrane sialoglycocomponents previously observed in various cell types (Pinto da Silva et al., 1981d;Torrisi and Pinto da Silva, 1984). This report demonstrates how the fracture-label technique is a powerful way to analyze membrane components.…”

Section: Introductionmentioning

confidence: 84%

Wheat germ agglutinin fracture‐label of lymphocyte plasma and intracellular membranes

Torrisi¹,

Pavan²,

Lotti³

et al. 1986

J. Elec. Microsc. Tech.

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“…The second useful advantage of fracture-label technique is its possible application to the analysis of intracellular membrane components, approaching problems related to their distribution, compartmentalization and transport (Torrisi and Pinto da Silva 1984;Torrisi et al 1989a). The method, in fact, exposes large areas of the intracellular membranes to the immunolabeling, providing full accessibility of the antigens; in the meantime, it preserves well the cellular ultrastructure and permits easy identification of the labeled organelles.…”

Section: Fracture-labelmentioning

confidence: 99%

Freeze-fracture immunogold labeling

Torrisi¹,

Mancini²

1996

Histochem Cell Biol

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Several approaches have been developed to combine immunogold cytochemistry and freeze-fracture techniques. These methods are highly heterogeneous regarding both the sequence of the procedural steps and the aspect of the resulting images. They imply immunolabeling either before or after freeze-fracture or even immunolabeling of platinum/carbon replicas of the freeze-fractured membranes, and have been used alternatively or in parallel to address different questions related to cell membrane structure, composition and dynamics or to intracellular membrane traffic. This review will briefly describe these methods and report most of their immunogold cytochemical applications, with the aim of facilitating selection of the most appropriate approach.

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“…In the Golgi apparatus-the organelle exclusively equipped with the enzymatic machinery for terminal glycosylation-weak labeling associated with forming secretory vesicles is apparent (C, arrows). This suggests that transfer of fully glycosylated products from Golgi to secretory granule membrane is rapid (Torrisi and Pinto da Silva, 1984). For further examples of the application of fracturelabel to intracellular membranes, see Aguas and Pinto da Silva (19851, Torrisi and Bonatti (1985), Torrisi et al (1987), and Coulombe et al (1988).…”

Section: Labeling Directly After Shadowingmentioning

confidence: 99%

Freeze‐fracture cytochemistry: Review of methods

Severs

1989

J. Elec. Microsc. Tech.

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"Freeze-fracture cytochemistry" encompasses a diversity of recently developed techniques in which freeze-fracture and cytochemistry are combined. Cytochemical labeling may, in principle, be integrated into one of three basic points in the standard freeze-fracture procedure; 1) before the specimen is frozen, 2) after it has been fractured, or 3) after it has been platinum shadowed and/or carbon coated. Visualization of the labeled cellular structures can be achieved by a variety of different methodologies. For example, the markers (usually colloidal gold particles) may be viewed embedded within a replica, or attached to it via fragments of membrane (or other cellular components). Sectioning is a central strategy in a number of techniques, either in combination with or in place of replication. The different combinations of methods that have been devised are not, for the most part, alternative ways of arriving at the same result; each provides quite distinct information about specific classes of membrane component or other structure in the cell. The purpose of this review is to present, within a single article, a systematic survey of the full range of techniques currently available in freeze-fracture cytochemistry. Emphasis is placed on explaining the principles underlying the methods and on illustrating their applications. With the success recently achieved, freeze-fracture cytochemistry has moved from the phase of experimental development to a position in which it may be expected increasingly to make significant contributions across a wide spectrum of problems in cell and membrane biology.

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Compartmentalization of intracellular membrane glycocomponents is revealed by fracture-label.

Cited by 28 publications

References 33 publications

Wheat germ agglutinin fracture‐label of lymphocyte plasma and intracellular membranes

Wheat germ agglutinin fracture‐label of lymphocyte plasma and intracellular membranes

Freeze-fracture immunogold labeling

Freeze‐fracture cytochemistry: Review of methods

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