Organotypic monolayer cultures of nervous tissue

Gähwiler, Beat H.

doi:10.1016/0165-0270(81)90003-0

Cited by 823 publications

(393 citation statements)

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“…Pictures also show that the architecture of the hippocampus was preserved even after 3 weeks in culture (Figure 2a), demonstrating that the interface culture method is optimal for slice maintenance, contrary to the roller tube technique, initially introduced by Gähwiler (1981), which can induce disorganization causing a monolayer aspect of tissues and the formation of holes.…”

Section: Discussionmentioning

confidence: 96%

“…This network has appropriate regional differentiation imitating the endogenous developmental changes in the hippocampus during the first few weeks after birth (Bahr, 1995; Muller, Buchs, & Stoppini, 1993). The culture of slices can be achieved through different procedures, such as the roller tube technique (Gähwiler, 1981) or the interface method, which maintains slices on a porous membrane at the interface between the culture medium and humidified air by using culture inserts. As such, the oxygenation of slices is optimized while providing adequate nutrition by capillarity (Stoppini, Buchs, & Muller, 1991).…”

Section: Introductionmentioning

confidence: 99%

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A new method allowing long‐term potentiation recordings in hippocampal organotypic slices

2017

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BackgroundHippocampal organotypic slices are used to improve the understanding of synaptic plasticity mechanisms because they allow longer term studies compared to acute slices. However, it is more delicate to keep cultures alive in the recording system outside in vitro conditions. Experiments from the organotypic cultures are common but the handling of slices is rarely described in the literature, even though tissue preservation is crucial. Instruments are sometimes required to extract the slices from the culture inserts but this approach is delicate and can lead to damage, given how strongly the slices are attached to the insert.MethodsA new configuration is proposed to secure the transfer of slices from the incubator to the recording chamber through an adaptor piece that can be designed for any model of chamber and/or insert. The adaptor is a Plexiglas ring in which a culture insert containing the slice can be easily introduced and stabilized. This system allows slices to be placed in the interface for electrophysiological investigations without having to detach them from the insert. That way, no damage is caused and the recording system can safely hold the slices, maintaining them close to culture conditions.ResultsIn addition to the description of the adaptation system, slices were characterized. Their viability was validated and microglial expression was observed. According to the experimental conditions, neuroprotective ramified microgliocytes are present. Dendritic spines studies were also performed to determine neuronal network maturity in culture. Moreover, SKF 83822 hydrobromide and three trains of 100 pulses at 100 Hz with a 10‐min inter‐train interval are suggested to induce long‐term potentiation and to record an increase of fEPSP amplitude and slope.ConclusionThis paper provides detailed information on the preparation and characterization of hippocampal organotypic slices, a new recording configuration more suitable for cultures, and a long‐term potentiation protocol combining SKF and trains.

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Section: Discussionmentioning

confidence: 96%

Section: Introductionmentioning

confidence: 99%

A new method allowing long‐term potentiation recordings in hippocampal organotypic slices

2017

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“…Avec cette technique, des tranches de différentes parties du système nerveux central de rongeurs nouveau-nés maintiennent leur organisation organotypique et permettent la visualisation des neurones. De plus, les cellules sont accessibles pour la micromanipulation et elles survivent plusieurs semaines, offrant alors la possibilité de les traiter avec différentes molécules [5][6][7]. Les explants peuvent aussi être spontanément fixés à une membrane poreuse et transparente afin d'être maintenus à l'interface air-liquide.…”

Section: Cultures De Tranches De Tissu Nerveuxunclassified

Modélisationin vitrodu système nerveux par génie tissulaire

2009

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“…The organotypic hippocampal culture technique allows for both the accessibility and long-term maintenance of an in vitro culture system, while maintaining a more physiological glial-neuronal relationship (Gahwiler 1981(Gahwiler & 1984Zimmer & Gahwiler 1984;Stoppini et al 1991). Several alternative techniques are currently available.…”

mentioning

confidence: 99%

“…Several alternative techniques are currently available. However, the ''thin slice culture'' technique relies on the use of comparatively thin tissue sections (Ͻ orΩ150 mm) as compared to those in other culture methods (250-500 mm) (Gahwiler 1981;Stoppini et al 1991;Song et al 1996;Parsley et al 1998). Thus, microscopic visualization and tissue oxygenation are enhanced.…”

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confidence: 99%

Impairment of the Ca²⁺‐Permeable AMPA/Kainate Receptors by Lead Exposure in Organotypic Rat Hippocampal Slice Cultures

Sui

Diyun²

2000

Pharmacology & Toxicology

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Previous studies have demonstrated that chronic lead exposure may impair neuronal process underlying synaptic plasticity via a direct interaction with N-methyl-D-aspartate (NMDA) receptors. The present study was carried out to investigate the effects of lead exposure on non-NMDA (a-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid, AMPA/ kainate) receptors of rat hippocampus. Ca 2π -permeable AMPA/kainate receptors in organotypic slice cultures were evaluated by using cobalt uptake, a histochemical method that identifies cells expressing Ca 2π -permeable non-NMDA receptors. Ten mM L-glutamate-induced cobalt accumulation was enriched in area CA1, area CA3 and in dentate gyrus, which was totally blocked by 100 mM DL-2-amino-5-phosphonovaleric acid (AP5) and 100 mM 6-cyano-7-nitroquinoxaline-2, 3-dione (CNQX). Three hundred mM NMDA-induced cobalt accumulation was in area CA1, area dentate gyrus and was blocked by AP5 or CNQX. One hundred mM AMPA had effects in area CA1, area CA3 and in dentate gyrus, which were blocked by CNQX, not by AP5. Furthermore, cobalt accumulations induced by NMDA and AMPA in the lead-exposed rats decreased significantly than those in the controls. The results indicate that AMPA receptors enriched in area CA1, area CA3, area dentate gyrus and kainate receptors enriched in area CA1, area dentate gyrus are impaired by lead exposure.

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Organotypic monolayer cultures of nervous tissue

Cited by 823 publications

References 20 publications

A new method allowing long‐term potentiation recordings in hippocampal organotypic slices

A new method allowing long‐term potentiation recordings in hippocampal organotypic slices

Modélisationin vitrodu système nerveux par génie tissulaire

Impairment of the Ca²⁺‐Permeable AMPA/Kainate Receptors by Lead Exposure in Organotypic Rat Hippocampal Slice Cultures

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Organotypic monolayer cultures of nervous tissue

Cited by 823 publications

References 20 publications

A new method allowing long‐term potentiation recordings in hippocampal organotypic slices

A new method allowing long‐term potentiation recordings in hippocampal organotypic slices

Modélisationin vitrodu système nerveux par génie tissulaire

Impairment of the Ca2+‐Permeable AMPA/Kainate Receptors by Lead Exposure in Organotypic Rat Hippocampal Slice Cultures

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Impairment of the Ca²⁺‐Permeable AMPA/Kainate Receptors by Lead Exposure in Organotypic Rat Hippocampal Slice Cultures