Context-Dependent Toxicity of Amyloid-β Peptides on Mouse Cerebellar Cells

Abstract

Alzheimer's disease (AD) is the major cause of dementia in old people. AD pathology is characterized by amyloid-β (Aβ) deposits in several regions of the brain, and links have been hypothesized between Aβ toxicity and apoptosis. Cerebellar granule cells (CGCs) have been widely used as in vitro tools for molecular studies correlating apoptosis with AD, although the cerebellum is a relatively spared area of the brain in vivo. We have used mixed neuronal-glial cerebellar cultures (NGCCs) and organotypic cerebellar cultures (OCCs) obtained from postnatal mice to assess the toxic effect of the Aβ oligomer 1-40 (Aβ1-40) after propidium iodide uptake in vitro. Our results demonstrate that NGCCs, which are primarily composed of CGCs, are resistant to Aβ1-40 challenge (5-10 μM) when cultured in physiological (5 mM) extracellular KCl ([K+]e) concentrations, i.e., in a condition in which CGCs undergo full maturation. Conversely, when 10 μM Aβ1-40 is given to NGCCs cultured in elevated (25 mM) [K+]e (and thus maintained in an immature state), there is a statistically significant increase in cell death. Cell death is by apoptosis, as demonstrated by ultrastructural examination. OCCs are resistant to Aβ challenge in any of the conditions tested (variation of [K+]e, presence or absence of serum, or addition of the neprilysin blocker phosphoramidon). Altogether these observations lead us to conclude that cerebellar cells in an organotypic context may be less susceptible to damage by Aβ, raising the question whether isolated CGCs are a reliable assay in drug discovery studies of AD.

STX, a Novel Membrane Estrogen Receptor Ligand, Protects Against Amyloid-β Toxicity

“…It is notable that we obtained similar results in the MC65 cells exposed to full-length Aβ peptides (Aβ 1–40/42 ) and SH-SY5Y cells exposed to the shorter peptides (Aβ 25–35 ) that have been shown to recapitulate the cytotoxic effects of Aβ oligomers [42, 58, 59]. Likewise, the coherence of our observations in both neuroblastoma lines and cultured neurons suggests that the protective effects of STX are not limited to a particular set of in vitro conditions [64]. In addition, our results are consistent with previous reports that STX is neuroprotective in whole-animal models of ischemic stroke [9, 63], supporting the hypothesis that this compound might be beneficial in the context of a variety of neurodegenerative conditions.…”

Unraveling the link: exploring the causal relationship between diabetes, multiple sclerosis, migraine, and Alzheimer’s disease through Mendelian randomization

“…Alasia et al cultured cerebellar granule cells in vitro and confirmed that Aβ deposition is the direct cause of brain cell apoptosis. At the same time, Aβ deposition also produces senile plaques and induces cognitive impairment in the body (Alasia et al, 2012). Disturbances in glucose metabolism have been shown to impact mitochondrial homeostasis in the brain (Cabezas-Opazo et al, 2015).…”

Fructose-1,6-Bisphosphate Protects Hippocampal Rat Slices from NMDA Excitotoxicity

“…Organotypic slice cultures represent a valid experimental model, maintaining neuronal maturation and plasticity, useful to evaluate experimental insults and drug treatments that can be followed for various days in vitro [31]. Using this model, toxicity of the Aβ oligomer 1–40, characteristic of the Alzheimer’s disease, has been evaluated in mixed neuronal-glial cerebellar cultures and organotypic cerebellar cultures [32]. Organotypic hippocampal slice cultures (OHSC) specifically uses hippocampal slices, mainly obtained from neonatal rats, to study the biochemical, morphological and functional characteristics of both neurons and glial cells.…”