Markers for neuronal degeneration in organotypic slice cultures

Noraberg, Jens; Kristensen, Bjarne Winther; Zimmer, Jens

doi:10.1016/s1385-299x(98)00050-6

Cited by 267 publications

(215 citation statements)

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“…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). In the presently used protocol, EPO and CEPO were applied for 24 h prior to the lesion, during OGD and 24 h thereafter or during the 24 h of NMDA exposure.…”

Section: Discussionmentioning

confidence: 99%

“…PI was added to the medium 24 h before OGD of control and experimental cultures to yield a 2 μM concentration. Recordings of cellular PI uptake by fluorescence microphotographs were performed immediately before the hypoxic or the excitotoxic insult and 24 h after using a tetramethyl rhodamine isothiocyanate filter (510-560/ 590 nm) and a Sensys KAF 1400 G2 (Photometrics, Tucson, AZ) digital camera (Noraberg et al, 1999). Correspondingly, timed recordings were performed from the respective controls.…”

Section: Determination Of the Et 50 Value For Ogd In Mouse Hippocampamentioning

confidence: 99%

“…One series was cell stained by toluidine blue to evaluate general morphology of the hippocampal slice cultures. The other series was immunostained for the microtubule-associated protein 2 (MAP2) to visualize neuronal cytoskeletal integrity (Noraberg et al, 1999). Changes in MAP2 staining were recorded by digital photographs of the whole hippocampus and CA1.…”

Section: Oxygen-glucose Deprivation (Ogd) Experimental Protocolmentioning

confidence: 99%

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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%

Section: Determination Of the Et 50 Value For Ogd In Mouse Hippocampamentioning

confidence: 99%

Section: Oxygen-glucose Deprivation (Ogd) Experimental Protocolmentioning

confidence: 99%

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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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“…no. 21103-049) with Hippocampal slice cultures were prepared as previously 25 mM D-glucose, 1 mM L-glutamine (Sigma, Vallensbaek described [20,30]. In brief, 5-7-day-old rat pups of Wistar Strand, Denmark, cat.…”

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confidence: 99%

“…The Timm sulphide-silver method, previously developed protocol known to be suitable for which histochemically detects and by its staining precipidetecting regional and quantitative differences [19,20,38] tate visualizes zinc associated in particular with gluta- (Fig. 3).…”

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Biocompatibility of silicon-based arrays of electrodes coupled to organotypic hippocampal brain slice cultures

et al. 2001

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In this study we examined the passive biocompatibility of a three-dimensional microelectrode array (MEA), designed to be coupled to organotypic brain slice cultures for multisite recording of electrophysiological signals. Hippocampal (and corticostriatal) brain slices from 1-week-old (and newborn) rats were grown for 4-8 weeks on the perforated silicon chips with silicon nitride surfaces and 40 mm sized holes and compared with corresponding tissue slices grown on conventional semiporous membranes. In terms of preservation of the basic cellular and connective organization, as visualized by Nissl staining, Timm sulphide silver-staining, microtubule-associated protein 2 (MAP2) and glial fibrillary acidic protein (GFAP) immunostaining, the slice cultures grown on chips did not differ from conventionally grown slice cultures. Neither were there any signs of astrogliosis or neurodegeneration around the upper recording part of the 47-mm-high platinum-tip electrodes. Slice cultures grown on a separate set of chips with platinum instead of silicon nitride surfaces also displayed normal MAP2 and GFAP immunostaining. The width of the GFAP-rich zone (glia limitans) at the bottom surface of the slice cultures was the same (|20 mm) in cultures grown on chips with silicon nitride and platinum surfaces and on conventional insert membranes. The slice cultures grown on chips maintained a normal, subfield differentiated susceptibility to the glutamate receptor agonist N-methyl-D-aspartate (NMDA) and the neurotoxin trimethyltin (TMT), as demonstrated by the cellular uptake of propidium iodide (PI), which was used as a reproducible and quantifiable marker for neuronal degeneration. We conclude that organotypic brain slice cultures can grow on silicon-based three-dimensional microelectrode arrays and develop normally with display of normal subfield differentiated susceptibilities to known excito-and neurotoxins. From this it is anticipated that the set-up, designed for recording of electrophysiological parameters, can be used for long-term studies of defined neuronal networks and provide valuable information on both normal, neurotoxicological and neuropathological conditions. Theme: Disorders of the nervous systemTopic: Neurotoxicity

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Physical Form Bright orange-yellow powder Solubility Freely soluble in water; soluble in ethanol, cellosolve; insoluble in xylene Melting Point 300 C Absorption (l max ) 425 nm Synthesis Synthetic methods 1-25 ,91 dermal toxicity; 92 genotoxicity; 93-96 hyperactive behavior in children; 97 mutagenicity; 98-106 neurotoxicity; 107 pseudoallergic reactions; 108 reproductive toxicity 107 Ruoff, M. K.; Vittum, P. W. The formation of azomethine dyes from 4-anisylideneand 4,4 0 -anisylidenebispyrazolones. J. Am. Chem. Soc. 1950, 72, 4947-4949. 21. Freeman, K. A.; Jones, J. H.; Graichen, C. Coal-tar colors. VIII. FD&C yellow No. 5. J. Assoc. Off.

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Markers for neuronal degeneration in organotypic slice cultures

Cited by 267 publications

References 39 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

Biocompatibility of silicon-based arrays of electrodes coupled to organotypic hippocampal brain slice cultures

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