Organotypic slice cultures: a technique has come of age

Gähwiler, Beat H.; Capogna, Marco; Debanne, Dominique; McKinney, R. Anne; Thompson, Scott M.

doi:10.1016/s0166-2236(97)01122-3

Cited by 809 publications

(638 citation statements)

References 92 publications

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“…Experimental in vitro models are often used to study biochemical and morphological changes related to degeneration processes because they allow a more easy and precise control of the extracellular environment compared to in vivo models. Organotypic hippocampal slice cultures offer several advantages in terms of preservation of 3-dimensional structure (Gahwiler, 1984;Stoppini et al, 1991) where the basic neuron-neuron and neuron-glia connections are maintained with normal electrophysiological properties (Blaabjerg et al, 2003;Frotscher et al, 1990;Gahwiler, 1988;Gahwiler et al, 1997;Zimmer and Gahwiler, 1984). For our experiments, mouse hippocampal slice cultures of postnatal brain tissue were established and grown on semiporous membranes, using the method of Stoppini et al (1991) as modified by Noraberg et al (1999).…”

Section: Discussionmentioning

confidence: 99%

Comparison of neuroprotective effects of erythropoietin (EPO) and carbamylerythropoietin (CEPO) against ischemia-like oxygen–glucose deprivation (OGD) and NMDA excitotoxicity in mouse hippocampal slice cultures

Montero

Poulsen

Noraberg

et al. 2007

Experimental Neurology

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In addition to its well-known hematopoietic effects, erythropoietin (EPO) also has neuroprotective properties. However, hematopoietic side effects are unwanted for neuroprotection, underlining the need for EPO-like compounds with selective neuroprotective actions. One such compound, devoid of hematopoietic bioactivity, is the chemically modified, EPO-derivative carbamylerythropoietin (CEPO). For comparison of the neuroprotective effects of CEPO and EPO, we subjected organotypic hippocampal slice cultures to oxygen-glucose deprivation (OGD) or N-methyl-D-aspartate (NMDA) excitotoxicity. Hippocampal slice cultures were pretreated for 24 h with 100 IU/ml EPO (= 26 nM) or 26 nM CEPO before OGD or NMDA lesioning. Exposure to EPO and CEPO continued during OGD and for the next 24 h until histology, as well as during the 24 h exposure to NMDA. Neuronal cell death was quantified by cellular uptake of propidium iodide (PI), recorded before the start of OGD and NMDA exposure and 24 h after. In cultures exposed to OGD or NMDA, CEPO reduced PI uptake by 49 ± 3 or 35 ± 8%, respectively, compared to lesion-only controls. EPO reduced PI uptake by 33 ± 5 and 15 ± 8%, respectively, in the OGD and NMDA exposed cultures. To elucidate a possible mechanism involved in EPO and CEPO neuroprotection against OGD, the integrity of α-II-spectrin cytoskeletal protein was studied. Both EPO and CEPO significantly reduced formation of spectrin cleavage products in the OGD model. We conclude that CEPO is at least as efficient neuroprotectant as EPO when excitotoxicity is modeled in mouse hippocampal slice cultures.

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Section: Discussionmentioning

confidence: 99%

Comparison of neuroprotective effects of erythropoietin (EPO) and carbamylerythropoietin (CEPO) against ischemia-like oxygen–glucose deprivation (OGD) and NMDA excitotoxicity in mouse hippocampal slice cultures

Montero

Poulsen

Noraberg

et al. 2007

Experimental Neurology

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“…Several methods can be used to prepare slice cultures (Gähwiler et al, 1997). Simple culture dishes will sustain slice cultures for a few days.…”

Section: Brain Slices In Culturementioning

confidence: 99%

Models of drug-induced epileptiform synchronization in vitro

Avoli

Jefferys

2016

Journal of Neuroscience Methods

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Models of epileptiform activity in vitro have many advantages for recording and experimental manipulation. Neural tissues can be maintained in vitro for hours, and in neuronal or organotypic slice cultures for several weeks. A variety of drugs and other agents increase activity in these in vitro conditions, in many cases resulting in epileptiform activity, thus providing a direct model of symptomatic seizures. We review these preparations and the experimental manipulations used to induce epileptiform activity. The most common of drugs used are GABA A receptor antagonists and potassium channel blockers (notably 4-aminopyridine). Muscarinic agents also can induce epileptiform synchronization in vitro, and include potassium channel inhibition amongst their cellular actions. Manipulations of extracellular ions are reviewed in another paper in this special issue, as are ex vivo slices prepared from chronically epileptic animals and from people with epilepsy. More complex slices including extensive networks and/or several connected brain structures can provide insights into the dynamics of long range connections during epileptic activity. Visualization of slices also provides opportunities for identification of living neurons and for optical recording/stimulation and manipulation. Overall, the analysis of the epileptiform activity induced in brain tissue in vitro has played a major role in advancing our understanding of the cellular and network mechanisms of epileptiform synchronization, and it is expected to continue to do so in future.

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“…The in vitro brain slice model may be lower throughput than the cell culture model, however, it is far more physiologically accurate. Within each slice, cytoarchitecture is maintained and thus many of the cell-to-cell interactions and neuronal networks remain intact (Gahwiler et al, 1997;Noraberg et al, 2005;Lossi et al, 2009). Hence, this model is well suited for physiological experiments to assess mechanism of action of drugs as well as to study neurophysiological changes that occur with stroke.…”

Section: E-mail Address: Tsaleh@upeica (Tm Saleh)mentioning

confidence: 99%

A novel method for inducing focal ischemia in vitro

Richard

Saleh

Bahh³

et al. 2010

Journal of Neuroscience Methods

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Current in vitro models of stroke involve applying oxygen-glucose deprived (OGD) media over an entire brain slice or plate of cultured neurons. Thus, these models fail to mimic the focal nature of stroke as observed clinically and with in vivo rodent models of stroke. Our aim was to develop a novel in vitro brain slice model of stroke that would mimic focal ischemia and thus allow for the investigation of events occurring in the penumbra. This was accomplished by focally applying OGD medium to a small portion of a brain slice while bathing the remainder of the slice with normal oxygenated media. This technique produced a focal infarct on the brain slice that increased as a function of time. Electrophysiological recordings made within the flow of the OGD solution ("core") revealed that neurons rapidly depolarized (anoxic depolarization; AD) in a manner similar to that observed in other stroke models. Edaravone, a known neuroprotectant, significantly delayed this onset of AD. Electrophysiological recordings made outside the flow of the OGD solution ("penumbra") revealed that neurons within this region progressively depolarized throughout the 75 min of OGD application. Edaravone attenuated this depolarization and doubled neuronal survival. Finally, synaptic transmission in the penumbra was abolished within 50 min of focal OGD application. These results suggest that this in vitro model mimics events that occur during focal ischemia in vivo and can be used to determine the efficacy of therapeutics that target neuronal survival in the core and/or penumbra.

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Organotypic slice cultures: a technique has come of age

Cited by 809 publications

References 92 publications

Comparison of neuroprotective effects of erythropoietin (EPO) and carbamylerythropoietin (CEPO) against ischemia-like oxygen–glucose deprivation (OGD) and NMDA excitotoxicity in mouse hippocampal slice cultures

Comparison of neuroprotective effects of erythropoietin (EPO) and carbamylerythropoietin (CEPO) against ischemia-like oxygen–glucose deprivation (OGD) and NMDA excitotoxicity in mouse hippocampal slice cultures

Models of drug-induced epileptiform synchronization in vitro

A novel method for inducing focal ischemia in vitro

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