Evaluation of cell damage in organotypic hippocampal slice culture from adult mouse: A potential model system to study neuroprotection

Su, Tao; Paradiso, Beatrice; Long, Yue-Sheng; Liao, Wei‐Ping; Simonato, Michele

doi:10.1016/j.brainres.2011.01.115

Cited by 42 publications

(43 citation statements)

References 38 publications

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“…We believe this process may negatively affect adult tissue, which is more sensitive to low temperatures, than tissue from young animals (1). We used PI staining to quantify the impact of sectioning temperature on section viability as previously performed by numerous groups on organotypic slice cultures (9,12,32,34,51). Overall, PI fluorescence of a section is linearly correlated with the number of dead cells within a particular section (34); thus, fluorescence intensity is an effective measure of section viability.…”

Section: Discussionmentioning

confidence: 99%

“…Brain sections were incubated with 10 mM propidium iodide (PI) for 10 min to assess cell death similar to that of previously published protocols (9,12,32,34,51). After this incubation, sections were rinsed twice with media, and PI fluorescence was imaged using ϫ10 objectives of a Nikon Ni-U upright fluorescence microscope (Nikon, Tokyo, Japan).…”

Section: Propidium Iodide Fluorescence Imagingmentioning

confidence: 99%

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Functional mitochondrial analysis in acute brain sections from adult rats reveals mitochondrial dysfunction in a rat model of migraine

Fried

Moffat

Seifert

et al. 2014

American Journal of Physiology-Cell Physiology

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Mitochondrial dysfunction has been implicated in many neurological disorders that only develop or are much more severe in adults, yet no methodology exists that allows for medium-throughput functional mitochondrial analysis of brain sections from adult animals. We developed a technique for quantifying mitochondrial respiration in acutely isolated adult rat brain sections with the Seahorse XF Analyzer. Evaluating a range of conditions made quantifying mitochondrial function from acutely derived adult brain sections from the cortex, cerebellum, and trigeminal nucleus caudalis possible. Optimization of this technique demonstrated that the ideal section size was 1 mm wide. We found that sectioning brains at physiological temperatures was necessary for consistent metabolic analysis of trigeminal nucleus caudalis sections. Oxygen consumption in these sections was highly coupled to ATP synthesis, had robust spare respiratory capacities, and had limited nonmitochondrial respiration, all indicative of healthy tissue. We demonstrate the effectiveness of this technique by identifying a decreased spare respiratory capacity in the trigeminal nucleus caudalis of a rat model of chronic migraine, a neurological disorder that has been associated with mitochondrial dysfunction. This technique allows for 24 acutely isolated sections from multiple brain regions of a single adult rat to be analyzed simultaneously with four sequential drug treatments, greatly advancing the ability to study mitochondrial physiology in adult neurological disorders.

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Section: Discussionmentioning

confidence: 99%

Section: Propidium Iodide Fluorescence Imagingmentioning

confidence: 99%

Functional mitochondrial analysis in acute brain sections from adult rats reveals mitochondrial dysfunction in a rat model of migraine

Fried

Moffat

Seifert

et al. 2014

American Journal of Physiology-Cell Physiology

View full text Add to dashboard Cite

show abstract

“…The electrical-activity maps evidenced that a large fraction of the overall slice area showed large spike amplitudes. Besides gradual decreases in slice network activity and single-unit activity partially caused by cell death (Stoppini et al, 1993), and besides temporal silence periods in neuron activity fluctuations, neuronal activity levels also were impacted by media changes or pH- and temperature variations (Su et al, 2011; Mewes et al, 2012). Any decreases in spike amplitude will entail difficulties in signal detection and assignment of the signals to the respective single-units.…”

Section: Discussionmentioning

confidence: 99%

Multiple Single-Unit Long-Term Tracking on Organotypic Hippocampal Slices Using High-Density Microelectrode Arrays

et al. 2016

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A novel system to cultivate and record from organotypic brain slices directly on high-density microelectrode arrays (HD-MEA) was developed. This system allows for continuous recording of electrical activity of specific individual neurons at high spatial resolution while monitoring at the same time, neuronal network activity. For the first time, the electrical activity patterns of single neurons and the corresponding neuronal network in an organotypic hippocampal slice culture were studied during several consecutive weeks at daily intervals. An unsupervised iterative spike-sorting algorithm, based on PCA and k-means clustering, was developed to assign the activities to the single units. Spike-triggered average extracellular waveforms of an action potential recorded across neighboring electrodes, termed “footprints” of single-units were generated and tracked over weeks. The developed system offers the potential to study chronic impacts of drugs or genetic modifications on individual neurons in slice preparations over extended times.

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“…Furthermore a new therapeutic approach can be tested in only a few weeks allowing a rapid screening. Recently also adolescent and adult rats have been used and even human post-mortem tissue slices have been kept alive for a maximum time of 1 week (Finley et al, 2004; Su et al, 2011; Fernandez-Bueno et al, 2012). …”

Section: Discussionmentioning

confidence: 99%

Modeling nigrostriatal degeneration in organotypic cultures, a new ex vivo model of Parkinson’s disease

et al. 2014

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Parkinson’s disease (PD) is the second most frequent neurodegenerative disorder afflicting 2% of the population older than 65 years worldwide. Recently, brain organotypic slices have been used to model neurodegenerative disorders, including PD. They conserve brain three-dimensional architecture, synaptic connectivity and its microenvironment. This model has allowed researchers a simple and rapid method to observe cellular interactions and mechanisms. In the present study, we developed an organotypic PD model from rat brains that includes all the areas involved in the nigrostriatal pathway in a single slice preparation, without using neurotoxins to induce the dopaminergic lesion. The mechanical transection of the nigrostriatal pathway obtained during slice preparation induced PD-like histopathology. Progressive nigrostriatal degeneration was monitored combining innovative approaches, such as diffusion tensor magnetic resonance imaging (DT-RMI) to follow fiber degeneration and mass spectrometry to quantify striatal dopamine content, together with bright-field and fluorescence microscopy imaging. A substantia nigra dopaminergic cell number decrease was observed by immunohistochemistry against rat tyrosine hydroxylase (TH) reaching 80% after 2 days in culture associated with a 30% decrease of striatal TH-positive fiber density, a 15% loss of striatal dopamine content quantified by mass spectrometry and a 70% reduction of nigrostriatal fiber fractional anisotropy quantified by DT-RMI. In addition, a significant decline of medium spiny neuron density was observed from days 7 to 16. These sagittal organotypic slices could be used to study the early stage of PD, namely dopaminergic degeneration, and the late stage of the pathology with dopaminergic and GABAergic neuron loss. This novel model might improve the understanding of PD and may represent a promising tool to refine the evaluation of new therapeutic approaches.

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Evaluation of cell damage in organotypic hippocampal slice culture from adult mouse: A potential model system to study neuroprotection

Cited by 42 publications

References 38 publications

Functional mitochondrial analysis in acute brain sections from adult rats reveals mitochondrial dysfunction in a rat model of migraine

Functional mitochondrial analysis in acute brain sections from adult rats reveals mitochondrial dysfunction in a rat model of migraine

Multiple Single-Unit Long-Term Tracking on Organotypic Hippocampal Slices Using High-Density Microelectrode Arrays

Modeling nigrostriatal degeneration in organotypic cultures, a new ex vivo model of Parkinson’s disease

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