Consistent and reproducible staining of glia by a modified Golgi–Cox method

Gull, Sidra; Ingrisch, Ina; Tausch, Svetlana; Witte, Otto W.; Schmidt, S.

doi:10.1016/j.jneumeth.2015.08.029

Cited by 15 publications

(11 citation statements)

References 49 publications

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“…To examine the morphology and swelling of astrocytes, a modified Golgi-Cox staining was used (Gull et al, 2015). Some brain slices were stained with FD Rapid GolgiStainTM Kit (FD Neurotechnologies, Inc., USA).…”

Section: Histologymentioning

confidence: 99%

Malignant astrocyte swelling and impaired glutamate clearance drive the expansion of injurious spreading depolarization foci

Menyhárt

Frank

Farkas

et al. 2020

Preprint

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Spreading depolarizations (SD) indicate infarct maturation and predict worse clinical outcome in ischemic stroke. We demonstrate here in rodents that brain edema formation upon ischemic stroke impairs astroglial glutamate clearance and increases the tissue area invaded by SD. The cytotoxic glutamate accumulation predisposes an extensive bulk of tissue for a yet undescribed simultaneous depolarization (SiD). We confirm in rat brain slices under hypo-osmotic stress that SiD is the pathological expansion of prior SD foci, is associated with astrocyte swelling and triggers oncotic neuron death. The blockade of astrocytic aquaporin-4 channels and Na+/K+/Cl- co-transporters, or volume-regulated anion channels mitigated slice edema, glutamate accumulation and SiD occurrence. Reversal of slice edema by hyperosmotic treatment counteracted glutamate accumulation and prevented SiD. In contrast, paralysis of astrocyte metabolism or inhibition of astrocyte glutamate uptake reproduced the SiD phenotype. We discuss our results in the light of evidence for SiD in the human cortex. Our results emphasize the need of preventive osmotherapy in ischemic stroke.

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Section: Histologymentioning

confidence: 99%

Malignant astrocyte swelling and impaired glutamate clearance drive the expansion of injurious spreading depolarization foci

Menyhárt

Frank

Farkas

et al. 2020

Preprint

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“…We also found that the tissue-protectant (cryoprotectant) solution (de Olmos et al, 1978 ; Watson et al, 1986 ) is very helpful to preserve tissue quality, reduce the staining background, and improve the attachment of sections to gelatin-coated slides. Minor additional steps can be added to our protocol to amend it for younger mice (Koyama and Tohyama, 2012 ) or to increase the staining for glia cells more than neurons (Ranjan and Mallick, 2012 ; Gull et al, 2015 ). Using our protocol, we found that initial brief fixation of brain samples with 4% PFA for 1 h followed by dd-H 2 O washing improves the staining for younger mice (Figure 6 ).…”

Section: Resultsmentioning

confidence: 99%

“…Of these, the Golgi-Cox method is considered to be most reliable in demonstrating dendritic arborization with a low background (Buell, 1982 ; Koyama, 2013 ). Many modifications of this method have been conducted and most aimed to increase its reliability (Angulo et al, 1996 ; Gibb and Kolb, 1998 ; Koyama and Tohyama, 2012 ), to reduce the required time (Ranjan and Mallick, 2010 ; Levine et al, 2013 ; Patro et al, 2013 ), and to increase the selectivity of neuronal vs. glial staining or vice versa (Ranjan and Mallick, 2012 ; Gull et al, 2015 ). Also, commercial kits have been developed for relatively fast constant Golgi staining.…”

Section: Introductionmentioning

confidence: 99%

Golgi-Cox Staining Step by Step

2016

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Golgi staining remains a key method to study neuronal morphology in vivo. Since most protocols delineating modifications of the original staining method lack details on critical steps, establishing this method in a laboratory can be time-consuming and frustrating. Here, we describe the Golgi-Cox staining in such detail that should turn the staining into an easily feasible method for all scientists working in the neuroscience field.

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“…For astrocytic staining, the manufacturer’s protocol was modified as described by Gull et al (2015). Briefly, isoflurane anesthetized animals were transcardially perfused with 4% formaldehyde in 0.1 M phosphate buffer (pH 7.4), brains were post-fixed for 4 days in the same fixative additionally containing 8% glutaraldehyde, submerged in a 1:1 mixture of solution A and B for 3 days at 26°C, transferred into solution C for 7 days at 4°C, sectioned on a cryostat (Leica CM3050 S, Germany) into 150μm coronal slices, mounted on Superfrost slides (Thermo Scientific, USA), stained in solutions D and E for 10 minutes, covered with Entellan (Merk, Darmstadt, Germany) and coded before quantitative analysis.…”

Section: Methodsmentioning

confidence: 99%

Experience-dependent structural plasticity in the adult brain: How the learning brain grows

Schmidt

Gull

Herrmann

et al. 2020

Preprint

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Volumetric magnetic resonance imaging studies have shown that intense learning can be associated with grey matter volume increases in the adult brain. The underlying mechanisms are poorly understood. Here we used monocular deprivation in rats to analyze the mechanisms underlying use-dependent grey matter increases. Optometry for quantification of visual acuity was combined with volumetric magnetic resonance imaging and microscopic techniques in longitudinal and cross-sectional studies. We found an increased spatial vision of the open eye which was associated with a transient increase in the volumes of the contralateral visual and lateral entorhinal cortex. In these brain areas dendrites of neurons elongated, and there was a strong increase in the number of spines, the targets of synapses, which was followed by spine maturation and partial pruning. Astrocytes displayed a transient pronounced swelling and underwent a reorganization of their processes. The use-dependent increase in grey matter corresponded predominantly to the swelling of the astrocytes. Experience-dependent increase in brain grey matter volume indicates a gain of structure plasticity with both synaptic and astrocyte remodeling.HighlightsPerception learning causes a transient increase in brain grey matter volume detectable by MRI.This learning results in pronounced changes of neuronal dendrites and an increase in the number of dendritic spines.Structural neuronal plasticity is associated with a reorganization and transient swelling of astrocytes.Brain volume and astrocyte volume return to baseline post-learning, with a persistent increase in the number of mature spines.

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Consistent and reproducible staining of glia by a modified Golgi–Cox method

Cited by 15 publications

References 49 publications

Malignant astrocyte swelling and impaired glutamate clearance drive the expansion of injurious spreading depolarization foci

Malignant astrocyte swelling and impaired glutamate clearance drive the expansion of injurious spreading depolarization foci

Golgi-Cox Staining Step by Step

Experience-dependent structural plasticity in the adult brain: How the learning brain grows

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