A Study of Extracellular Space in Central Nervous Tissue by Freeze-Substitution

Harreveld, A. Van; Crowell, Jane; Malhotra, S. K.

doi:10.1083/jcb.25.1.117

Cited by 390 publications

(163 citation statements)

References 32 publications

(64 reference statements)

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“…Early studies by Van Harreveld and co-workers (Vanharreveld and Crowell, 1964;Vanharreveld et al, 1965) using a cold metal mirror freezing device demonstrated that brain tissue needed to be fast frozen in 30 sec after excision from a mouse and before the onset of cardiac arrest (an ~8 minute times scale). After this time, morphological deterioration was observed and it was reported that the tissue resembled chemically fixed specimens.…”

Section: Discussionmentioning

confidence: 99%

The combination of chemical fixation procedures with high pressure freezing and freeze substitution preserves highly labile tissue ultrastructure for electron tomography applications

Sosinsky

Crum

Jones

et al. 2008

Journal of Structural Biology

113

114

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The emergence of electron tomography as a tool for three dimensional structure determination of cells and tissues has brought its own challenges for the preparation of thick sections. High pressure freezing in combination with freeze substitution provides the best method for obtaining the largest volume of well-preserved tissue. However, for deeply embedded, heterogeneous, labile tissues needing careful dissection, such as brain, the damage due to anoxia and excision before cryofixation is significant. We previously demonstrated that chemical fixation prior to high pressure freezing preserves fragile tissues and produces superior tomographic reconstructions compared to equivalent tissue preserved by chemical fixation alone. Here, we provide further characterization of the technique, comparing the ultrastructure of Flock House Virus infected DL1 insect cells that were 1) high pressure frozen without fixation, 2) high pressure frozen following fixation, and 3) conventionally prepared with aldehyde fixatives. Aldehyde fixation prior to freezing produces ultrastructural preservation superior to that obtained through chemical fixation alone that is close to that obtained when cells are fast frozen without fixation. We demonstrate using a variety of nervous system tissues, including neurons that were injected with a fluorescent dye and then photooxidized, that this technique provides excellent preservation compared to chemical fixation alone and can be extended to selectively stained material where cryofixation is impractical.

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Section: Discussionmentioning

confidence: 99%

The combination of chemical fixation procedures with high pressure freezing and freeze substitution preserves highly labile tissue ultrastructure for electron tomography applications

Sosinsky

Crum

Jones

et al. 2008

Journal of Structural Biology

113

114

View full text Add to dashboard Cite

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“…*P , 0.05; ***P , 0.001. extracellular space consists of thin layers of interstitial medium separating neurons and glial cells, so that the synaptic cleft and its surroundings are mostly sealed from the chemical standpoint. The width of the thin sheets composing the interstitial extracellular space has been measured in the range of a few tens of nanometers (10-40 nm) (26)(27)(28). In choosing our model geometry, we aimed to capture the hindrance of diffusion in the perisynaptic extracellular space (29) and the relative compartmentalization of the extracellular space (30), but it is impossible to provide a stereotypic description of the interstitial space given that it exhibits great variability (26).…”

Section: Electrodiffusion Computational Modelingmentioning

confidence: 99%

“…We modeled electrodiffusion in the extracellular space according to Poisson-Nernst-Planck equations (24) and based the geometry of the synaptic surroundings on the robust observation that free extracellular space consists of a connected network of thin layers running between neurons and glial cells (26,27) (Fig. 7A, B).…”

Section: Vol 29 October 2015mentioning

confidence: 99%

Allosteric modulation of metabotropic glutamate receptors by chloride ions

et al. 2015

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Metabotropic glutamate receptors (mGluRs) play key roles in the modulation of many synapses. Chloride (Cl 2 ) is known to directly bind and regulate the function of different actors of neuronal activity, and several studies have pointed to the possible modulation of mGluRs by Cl 2 . Herein, we demonstrate that Cl 2 behaves as a positive allosteric modulator of mGluRs. For example, whereas glutamate potency was 3.08 6 0.33 mM on metabotropic glutamate (mGlu) 4 receptors in high-Cl 2 buffer, signaling activity was almost abolished in low Cl 2 in cell-based assays.

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“…Wide dissemination in extracellular spaces is an efficient and rapid mechanism of virus dissemination within the CNS but is more important in rapid progression of fixed strain of the rabies virus [51].…”

Section: Viral Spread To the Cnsmentioning

confidence: 99%

Perspectives in Diagnosis and Treatment of Rabies Viral Encephalitis: Insights from Pathogenesis

et al. 2016

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Rabies viral encephalitis, though one of the oldest recognized infectious disease of humans, remains an incurable, fatal encephalomyelitis, despite advances in understanding of its pathobiology. Advances in science have led us on the trail of the virus in the host, but the sanctuaries in which the virus remains hidden for its survival are unknown. Insights into host-pathogen interactions have facilitated evolving immunologic therapeutic strategies, though we are far from a cure. Most of the present-day knowledge has evolved from in vitro studies using fixed (attenuated) laboratory strains that may not be applicable in the clinical setting. Much remains to be unraveled about this elusive virus. This review attempts to re-examine the current advances in understanding of the pathobiology of the rabies virus that modulate the diagnosis, treatment, and prevention of this fatal disease.

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A Study of Extracellular Space in Central Nervous Tissue by Freeze-Substitution

Abstract: It was attempted to preserve the water distribution in central nervous tissue by rapid freezing followed by substitution fixation at low temperature.

Cited by 390 publications

References 32 publications

The combination of chemical fixation procedures with high pressure freezing and freeze substitution preserves highly labile tissue ultrastructure for electron tomography applications

The combination of chemical fixation procedures with high pressure freezing and freeze substitution preserves highly labile tissue ultrastructure for electron tomography applications

Allosteric modulation of metabotropic glutamate receptors by chloride ions

Perspectives in Diagnosis and Treatment of Rabies Viral Encephalitis: Insights from Pathogenesis

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