Organotypic brain slice cultures to model neurodegenerative proteinopathies

AbstractOrganotypic slice cultures of brain or spinal cord have been a longstanding tool in neuroscience research but their utility for understanding Alzheimer’s disease (AD) and other neurodegenerative proteinopathies has only recently begun to be evaluated. Organotypic brain slice cultures (BSCs) represent a physiologically relevant three-dimensional model of the brain. BSCs support all the central nervous system (CNS) cell types and can be produced from brain areas involved in neurodegenerative disease. BSCs can be used to better understand the induction and significance of proteinopathies underlying the development and progression of AD and other neurodegenerative disorders, and in the future may serve as bridging technologies between cell culture and in vivo experiments for the development and evaluation of novel therapeutic targets and strategies. We review the initial development and general use of BSCs in neuroscience research and highlight the advantages of these cultures as an ex vivo model. Subsequently we focus on i) BSC-based modeling of AD and other neurodegenerative proteinopathies ii) use of BSCs to understand mechanisms underlying these diseases and iii) how BSCs can serve as tools to screen for suitable therapeutics prior to in vivo investigations. Finally, we will examine i) open questions regarding the use of such cultures and ii) how emerging technologies such as recombinant adeno-associated viruses (rAAV) may be combined with these models to advance translational research relevant to neurodegenerative disorders.

Download Full-text

Preparation of organotypic brain slice cultures for the study of Alzheimer’s disease

F1000Research ◽

10.12688/f1000research.14500.1 ◽

2018 ◽

Vol 7 ◽

pp. 592

Author(s):

Cara L. Croft ◽

Wendy Noble

Keyword(s):

Alzheimer’S Disease ◽

Alzheimer's Disease ◽

Brain Slice ◽

Synaptic Dysfunction ◽

Slice Culture ◽

Therapeutic Effects ◽

Culture Model ◽

Organotypic Brain Slice ◽

Brain Slice Cultures

Alzheimer's disease, the most common cause of dementia, is a progressive neurodegenerative disorder characterised by amyloid-beta deposits in extracellular plaques, intracellular neurofibrillary tangles of aggregated tau, synaptic dysfunction and neuronal death. There are no cures for AD and current medications only alleviate some disease symptoms. Transgenic rodent models to study Alzheimer’s mimic features of human disease such as age-dependent accumulation of abnormal beta-amyloid and tau, synaptic dysfunction, cognitive deficits and neurodegeneration. These models have proven vital for improving our understanding of the molecular mechanisms underlying AD and for identifying promising therapeutic approaches. However, modelling neurodegenerative disease in animals commonly involves aging animals until they develop harmful phenotypes, often coupled with invasive procedures. In vivo studies are also resource, labour, time and cost intensive. We have developed a novel organotypic brain slice culture model to study Alzheimer’ disease which brings the potential of substantially reducing the number of rodents used in dementia research from an estimated 20,000 per year. We obtain 36 brain slices from each mouse pup, considerably reducing the numbers of animals required to investigate multiple stages of disease. This tractable model also allows the opportunity to modulate multiple pathways in tissues from a single animal. We believe that this model will most benefit dementia researchers in the academic and drug discovery sectors. We validated the slice culture model against aged mice, showing that the molecular phenotype closely mimics that displayed in vivo, albeit in an accelerated timescale. We showed beneficial outcomes following treatment of slices with agents previously shown to have therapeutic effects in vivo, and we also identified new mechanisms of action of other compounds. Thus, organotypic brain slice cultures from transgenic mouse models expressing Alzheimer’s disease-related genes may provide a valid and sensitive replacement for in vivo studies that do not involve behavioural analysis.

Download Full-text

A Bridge Between in vitro and in vivo Studies in Neuroscience: Organotypic Brain Slice Cultures

Nöro Psikiyatri Arşivi ◽

10.29399/npa.26139 ◽

2020 ◽

Author(s):

Merve Alaylioglu ◽

Erdinc Dursun ◽

Selma Yilmazer ◽

Duygu Gezen-Ak

Keyword(s):

Brain Slice ◽

In Vivo Studies ◽

Organotypic Brain Slice ◽

Brain Slice Cultures

Download Full-text

Preparation of organotypic brain slice cultures for the study of Alzheimer’s disease

F1000Research ◽

10.12688/f1000research.14500.2 ◽

2018 ◽

Vol 7 ◽

pp. 592 ◽

Cited By ~ 13

Author(s):

Cara L. Croft ◽

Wendy Noble

Keyword(s):

Alzheimer’S Disease ◽

Alzheimer's Disease ◽

Brain Slice ◽

Synaptic Dysfunction ◽

Slice Culture ◽

Therapeutic Effects ◽

Culture Model ◽

Organotypic Brain Slice ◽

Brain Slice Cultures

Alzheimer's disease, the most common cause of dementia, is a progressive neurodegenerative disorder characterised by amyloid-beta deposits in extracellular plaques, intracellular neurofibrillary tangles of aggregated tau, synaptic dysfunction and neuronal death. Transgenic rodent models to study Alzheimer’s mimic features of human disease such as age-dependent accumulation of abnormal beta-amyloid and tau, synaptic dysfunction, cognitive deficits and neurodegeneration. These models have proven vital for improving our understanding of the molecular mechanisms underlying AD and for identifying promising therapeutic approaches. However, modelling neurodegenerative disease in animals commonly involves aging animals until they develop harmful phenotypes, often coupled with invasive procedures. We have developed a novel organotypic brain slice culture model to study Alzheimer’s disease using 3xTg-AD mice which brings the potential of substantially reducing the number of rodents used in dementia research from an estimated 20,000 per year. Using a McIllwain tissue chopper, we obtain 36 x 350 micron slices from each P8-P9 mouse pup for culture between 2 weeks and 6 months on semi-permeable 0.4 micron pore membranes, considerably reducing the numbers of animals required to investigate multiple stages of disease. This tractable model also allows the opportunity to modulate multiple pathways in tissues from a single animal. We believe that this model will most benefit dementia researchers in the academic and drug discovery sectors. We validated the slice culture model against aged mice, showing that the molecular phenotype closely mimics that displayed in vivo, albeit in an accelerated timescale. We showed beneficial outcomes following treatment of slices with agents previously shown to have therapeutic effects in vivo, and we also identified new mechanisms of action of other compounds. Thus, organotypic brain slice cultures from transgenic mouse models expressing Alzheimer’s disease-related genes may provide a valid and sensitive replacement for in vivo studies that do not involve behavioural analysis.

Download Full-text

Oxidative Stress-Tolerant Stem Cells from Human Exfoliated Deciduous Teeth Decrease Hydrogen Peroxide-Induced Damage in Organotypic Brain Slice Cultures from Adult Mice

International Journal of Molecular Sciences ◽

10.3390/ijms20081858 ◽

2019 ◽

Vol 20 (8) ◽

pp. 1858 ◽

Cited By ~ 3

Author(s):

Li Xiao ◽

Chikako Saiki ◽

Hisashi Okamura

Keyword(s):

Oxidative Stress ◽

Hydrogen Peroxide ◽

Stem Cells ◽

Neurodegenerative Disorders ◽

Neurotrophic Factor ◽

Brain Slice ◽

Deciduous Teeth ◽

Organotypic Brain Slice ◽

Brain Slice Cultures ◽

Human Exfoliated Deciduous Teeth

Oxidative stress causes severe tissue injury of the central nervous system in ischemic brain damage (IBD), traumatic brain injury (TBI) and neurodegenerative disorders. In this study, we used hydrogen peroxide (H2O2) to induce oxidative stress in organotypic brain slice cultures (OBSCs), and investigated the protective effects of oxidative stress-tolerant (OST) stem cells harvested from human exfoliated deciduous teeth (SHED) which were co-cultivated with OBSCs. Using presto blue assay and immunostaining, we demonstrated that both normal SHED and OST-SHED could prevent H2O2-induced cell death, and increase the numbers of mature neuron and neuronal progenitors in the hippocampus of OBSCs. During co-cultivation, OST-SHED, but not normal SHED, exhibited neuronal cell morphology and expressed neuronal markers. Results from ELISA showed that both normal SHED and OST-SHED significantly decreased oxidative DNA damage in H2O2-treated OBSCs. SHED could also produce neurotrophic factor BDNF (brain derived neurotrophic factor) and promoted the production of IL-6 in OBSCs. Although OST-SHED had lower cell viability, the neuronal protection of OST-SHED was significantly superior to that of normal SHED. Our findings suggest that SHED, especially OST-SHED, could prevent oxidative stress induced brain damage. OST-SHED can be explored as a new therapeutic tool for IBD, TBI and neurodegenerative disorders.

Download Full-text

Neuronal activity modulates alpha-synuclein aggregation and spreading in organotypic brain slice cultures and in vivo

Acta Neuropathologica ◽

10.1007/s00401-020-02227-6 ◽

2020 ◽

Vol 140 (6) ◽

pp. 831-849

Author(s):

Qihui Wu ◽

Muhammad A. Shaikh ◽

Emily S. Meymand ◽

Bin Zhang ◽

Kelvin C. Luk ◽

...

Keyword(s):

Neuronal Activity ◽

Brain Slice ◽

Alpha Synuclein ◽

Organotypic Brain Slice ◽

Brain Slice Cultures ◽

Alpha Synuclein Aggregation

Download Full-text

Organotypic Brain Slices of ADULT Transgenic Mice: A Tool to Study Alzheimer’s Disease

Current Alzheimer Research ◽

10.2174/1567205016666181212153138 ◽

2019 ◽

Vol 16 (2) ◽

pp. 172-181

Author(s):

Christian Humpel

Keyword(s):

Alzheimer’S Disease ◽

Alzheimer's Disease ◽

Transgenic Mice ◽

Brain Slice ◽

Ex Vivo ◽

Brain Slices ◽

Adult Mice ◽

Organotypic Brain Slice ◽

Brain Slice Cultures ◽

Organotypic Brain Slices

Transgenic mice have been extensively used to study the Alzheimer pathology. In order to reduce, refine and replace (3Rs) the number of animals, ex vivo cultures are used and optimized. Organotypic brain slices are the most potent ex vivo slice culture models, keeping the 3-dimensional structure of the brain and being closest to the in vivo situation. Organotypic brain slice cultures have been used for many decades but were mainly prepared from postnatal (day 8-10) old rats or mice. More recent work (including our lab) now aims to culture organotypic brain slices from adult mice including transgenic mice. Especially in Alzheimer´s disease research, brain slices from adult transgenic mice will be useful to study beta-amyloid plaques, tau pathology and glial activation. This review will summarize the studies using organotypic brain slice cultures from adult mice to mimic Alzheimer's disease and will highlight advantages and also pitfalls using this technique.

Download Full-text

Organotypic Brain Slice Culture Microglia Exhibit Molecular Similarity to Acutely-Isolated Adult Microglia and Provide a Platform to Study Neuroinflammation

Frontiers in Cellular Neuroscience ◽

10.3389/fncel.2020.592005 ◽

2020 ◽

Vol 14 ◽

Author(s):

Alex R. D. Delbridge ◽

Dann Huh ◽

Margot Brickelmaier ◽

Jeremy C. Burns ◽

Chris Roberts ◽

...

Keyword(s):

Gene Expression ◽

Brain Slice ◽

Initial Period ◽

Slice Culture ◽

Organotypic Brain Slice ◽

Brain Slice Cultures ◽

The Impact ◽

Over Time

Microglia are central nervous system (CNS) resident immune cells that have been implicated in neuroinflammatory pathogenesis of a variety of neurological conditions. Their manifold context-dependent contributions to neuroinflammation are only beginning to be elucidated, which can be attributed in part to the challenges of studying microglia in vivo and the lack of tractable in vitro systems to study microglia function. Organotypic brain slice cultures offer a tissue-relevant context that enables the study of CNS resident cells and the analysis of brain slice microglial phenotypes has provided important insights, in particular into neuroprotective functions. Here we use RNA sequencing, direct digital quantification of gene expression with nCounter® technology and targeted analysis of individual microglial signature genes, to characterize brain slice microglia relative to acutely-isolated counterparts and 2-dimensional (2D) primary microglia cultures, a widely used in vitro surrogate. Analysis using single cell and population-based methods found brain slice microglia exhibited better preservation of canonical microglia markers and overall gene expression with stronger fidelity to acutely-isolated adult microglia, relative to in vitro cells. We characterized the dynamic phenotypic changes of brain slice microglia over time, after plating in culture. Mechanical damage associated with slice preparation prompted an initial period of inflammation, which resolved over time. Based on flow cytometry and gene expression profiling we identified the 2-week timepoint as optimal for investigation of microglia responses to exogenously-applied stimuli as exemplified by treatment-induced neuroinflammatory changes observed in microglia following LPS, TNF and GM-CSF addition to the culture medium. Altogether these findings indicate that brain slice cultures provide an experimental system superior to in vitro culture of microglia as a surrogate to investigate microglia functions, and the impact of soluble factors and cellular context on their physiology.

Download Full-text

Ex vivo therapy of malignant melanomas transplanted into organotypic brain slice cultures using inhibitors of histone deacetylases

Acta Neuropathologica ◽

10.1007/s00401-006-0082-8 ◽

2006 ◽

Vol 112 (2) ◽

pp. 205-215 ◽

Cited By ~ 11

Author(s):

Annett Hölsken ◽

Ilker Y. Eyüpoglu ◽

Mike Lueders ◽

Christian Tränkle ◽

Detlef Dieckmann ◽

...

Keyword(s):

Histone Deacetylases ◽

Brain Slice ◽

Ex Vivo ◽

Malignant Melanomas ◽

Organotypic Brain Slice ◽

Brain Slice Cultures

Download Full-text

Neuroprotective Effects of the FGF21 Analogue LY2405319

Journal of Alzheimer s Disease ◽

10.3233/jad-200837 ◽

2021 ◽

pp. 1-13

Author(s):

Claire Rühlmann ◽

David Dannehl ◽

Marcus Brodtrück ◽

Andrew C. Adams ◽

Jan Stenzel ◽

...

Keyword(s):

Glial Cells ◽

Neuroprotective Effect ◽

Amyloid Β ◽

Fibroblast Growth Factor 21 ◽

Neuroprotective Effects ◽

Organotypic Brain Slice ◽

Brain Slice Cultures ◽

Abca1 Mrna

Background: To date, there are no effective treatments for Alzheimer’s disease (AD). Thus, a significant need for research of therapies remains. Objective: One promising pharmacological target is the hormone fibroblast growth factor 21 (FGF21), which is thought to be neuroprotective. A clinical candidate for medical use could be the FGF21 analogue LY2405319 (LY), which has a specificity and potency comparable to FGF21. Methods: The present study investigated the potential neuroprotective effect of LY via PPARγ/apoE/abca1 pathway which is known to degrade amyloid-β (Aβ) plaques by using primary glial cells and hippocampal organotypic brain slice cultures (OBSCs) from 30- and 50-week-old transgenic APPswe/PS1dE9 (tg) mice. By LY treatment of 52-week-old tg mice with advanced Aβ deposition, we further aimed to elaborate the effect of LY on AD pathology in vivo. Results: LY application to primary glial cells caused an upregulation of pparγ, apoE, and abca1 mRNA expression and significantly decreased number and area of Aβ plaques in OBSCs. LY treatment in tg mice increased cerebral [18F] FDG uptake and N-acetylaspartate/creatine ratio indicating enhanced neuronal activity and integrity. Although LY did not reduce the number of Aβ plaques in tg mice, the number of iba1-positive cells was significantly decreased indicating reduced microgliosis. Conclusion: These data identified LY in vitro as an activator of Aβ degrading genes leading to cerebral Aβ load amelioration in early and late AD pathology. Although Aβ plaque reduction by LY failed in vivo, LY may be used as therapeutic agent to treat AD-related neuroinflammation and impaired neuronal integrity.

Download Full-text