Freeze-fracture immunogold labeling

Torrisi, Maria Rosaria; Mancini, Patrizia

doi:10.1007/bf02473199

Cited by 13 publications

(3 citation statements)

References 39 publications

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“…The resulting fracture faces do not contain the entire set of membrane proteins. Therefore, SDS-FRL, in some instances, may be limited by membrane protein partitioning, as described previously (Fujimoto 1995) and also pointed out by Torrisi and Mancini (1996). However, from other points of view, this preferential protein partitioning enables the precise topological analysis of integral membrane proteins by SDS-FRL using site-directed antibodies.…”

Section: Differential Phospholipid Analysis Of Protoplasmic and Exoplmentioning

confidence: 72%

SDS-digested freeze-fracture replica labeling electron microscopy to study the two-dimensional distribution of integral membrane proteins and phospholipids in biomembranes: practical procedure, interpretation and application

Fujimoto

1997

Histochemistry and Cell Biology

120

110

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Recently, we have developed a quick-freezing/freeze-fracture replica labeling technique, sodium dodecyl sulfate (SDS)-digested freeze-fracture replica labeling (SDS-FRL), to study the two-dimensional distribution of cytochemical labeling on the membrane surface and the relationship of this distribution to images of freeze-fracture replicas created by platinum shadowing. In SDS-FRL, unfixed, quick-frozen cells, after freeze-fracture and platinum/carbon shadowing, are treated with SDS. The detergent dissolves unfractured areas of the cell membranes, with the release of the cytoplasmic contents. The cytoplasmic and exoplasmic membrane surfaces can be then labeled cytochemically. Integral membrane proteins, revealed as intramembrane particles by freeze-fracture replication, which are indistinguishable on a purely morphological basis, can be selectively labeled by SDS-FRL with specific antibody. In addition, this approach can be applied to examine the transmembrane phospholipid distribution in various cell and intracellular membranes. In this review, we describe the practical procedure for SDS-FRL in detail, present its application to labeling of various membrane components, and briefly discuss the possibility of a combination of SDS-FRL with atomic force microscopy.

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Section: Differential Phospholipid Analysis Of Protoplasmic and Exoplmentioning

confidence: 72%

SDS-digested freeze-fracture replica labeling electron microscopy to study the two-dimensional distribution of integral membrane proteins and phospholipids in biomembranes: practical procedure, interpretation and application

Fujimoto

1997

Histochemistry and Cell Biology

120

110

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“…Since all these methods offer complementary information on protein localization, a detailed comprehensive assessment of receptors and transporter distribution at the cellular, subcellular and subsynaptic levels is usually achieved through a combination of techniques. Other promising high-resolution approaches that combine immunogold and freeze-fracture techniques (Torrisi and Mancini, 1996), have recently been used to study localization of receptor proteins (e. g. Hagiwara et al, 2005;Tanaka et al, 2005;Kulik et al, 2006).…”

Section: Postembedding Immunogoldmentioning

confidence: 99%

Glutamate and GABA receptors and transporters in the basal ganglia: What does their subsynaptic localization reveal about their function?

2006

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GABA and glutamate, the main transmitters in the basal ganglia, exert their effects through ionotropic and metabotropic receptors. The dynamic activation of these receptors in response to released neurotransmitter depends, among other factors, on their precise localization in relation to corresponding synapses. The use of high resolution quantitative electron microscope immunocytochemical techniques has provided in-depth description of the subcellular and subsynaptic localization of these receptors in the CNS. In this article, we review recent findings on the ultrastructural localization of GABA and glutamate receptors and transporters in the basal ganglia, at synaptic, extrasynaptic and presynaptic sites. The anatomical evidence supports numerous potential locations for receptor-neurotransmitter interactions, and raises important questions regarding mechanisms of activation and function of synaptic versus extrasynaptic receptors in the basal ganglia.

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“…3). The principle of the immunological detection of cellular constituents was extended by Bendayan (1981) Torrisi and Mancini 1996). An improvement for the two-dimensional analysis of cell surface receptor distribution was reported by Tolson et al (1981) who adapted the shadow cast replica technique in their studies on the distribution of the epidermal growth factor receptors.…”

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confidence: 98%

The silver anniversary of gold: 25 years of the colloidal gold marker system for immunocytochemistry and histochemistry

Roth

1996

Histochem Cell Biol

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Since 1971, when W.P. Faulk and G.M. Taylor published "An immunocolloid method for the electron microscope", colloidal gold has become a very widely used marker in microscopy. It has been used to detect a huge range of cellular and extracellular constituents by in situ hybridization, immunogold, lectin-gold, and enzyme-gold labeling. Besides its use in light microscopic immunogold and lectin-gold silver staining, colloidal gold remains the label of choice for transmission electron microscopy studying thin sections, freeze-etch, and surface replicas, as well as for scanning electron microscopy. The year 1996 is the 25th anniversary of the introduction of colloidal gold as a marker in immunoelectron microscopy and this overview outlines some of the major milestones in the development of the colloidal gold marker system.

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Freeze-fracture immunogold labeling

Cited by 13 publications

References 39 publications

SDS-digested freeze-fracture replica labeling electron microscopy to study the two-dimensional distribution of integral membrane proteins and phospholipids in biomembranes: practical procedure, interpretation and application

SDS-digested freeze-fracture replica labeling electron microscopy to study the two-dimensional distribution of integral membrane proteins and phospholipids in biomembranes: practical procedure, interpretation and application

Glutamate and GABA receptors and transporters in the basal ganglia: What does their subsynaptic localization reveal about their function?

The silver anniversary of gold: 25 years of the colloidal gold marker system for immunocytochemistry and histochemistry

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