Neurotoxic Ca2+ Signaling Induced by Amyloid–β Oligomers in Aged Hippocampal Neurons In Vitro

Núñez, Lucı́a; Calvo-Rodríguez, María; Caballero, Erica; García-Durillo, Mónica; Alonso, Carlos

doi:10.1007/978-1-4939-7816-8_20

Cited by 5 publications

(3 citation statements)

References 15 publications

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“…Interestingly, this effect of Aβ42 oligomer application was not revealed at the early stages of primary neuronal culture development (2-10 DIV), but Ca 2+ activity was sharply increased beginning at DIV 14, at which point different age-related signs were observed in neurons. In this regard, the cytosolic and mitochondrial Ca 2+ responses induced by single Aβ42 oligomer administration increased dramatically over time (adult cultures DIV 21-28) ( Calvo-Rodríguez et al, 2016 ; Núñez et al, 2018 ). Our data indicated that chronic Aβ42 application increased the suppression of functional Ca 2+ activity in primary hippocampal cultures and may correlate with in vivo studies of Ca 2+ activity, indicating neural dysfunction in the brains of aging mice with modeled AD.…”

Section: Discussionmentioning

confidence: 99%

Brain-Derived Neurotrophic Factor (BDNF) Preserves the Functional Integrity of Neural Networks in the β-Amyloidopathy Model in vitro

Митрошина

Yarkov

Mishchenko

et al. 2020

Front. Cell Dev. Biol.

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Alzheimer’s disease (AD) is a widespread chronic neurodegenerative pathology characterized by synaptic dysfunction, partial neuronal death, cognitive decline and memory impairments. The major hallmarks of AD are extracellular senile amyloid plaques formed by various types of amyloid proteins (Aβ) and the formation and accumulation of intracellular neurofibrillary tangles. However, there is a lack of relevant experimental models for studying changes in neural network activity, the features of intercellular signaling or the effects of drugs on the functional activity of nervous cells during AD development. In this work, we examined two experimental models of amyloidopathy using primary hippocampal cultures. The first model involves the embryonic brains of 5xFAD mice; the second uses chronic application of amyloid beta 1-42 (Aβ1-42). The model based on primary hippocampal cells obtained from 5xFAD mice demonstrated changes in spontaneous network calcium activity characterized by a decrease in the number of cells exhibiting Ca 2+ activity, a decrease in the number of Ca 2+ oscillations and an increase in the duration of Ca 2+ events from day 21 of culture development in vitro . Chronic application of Aβ1-42 resulted in the rapid establishment of significant neurodegenerative changes in primary hippocampal cultures, leading to marked impairments in neural network calcium activity and increased cell death. Using this model and multielectrode arrays, we studied the influence of amyloidopathy on spontaneous bioelectrical neural network activity in primary hippocampal cultures. It was shown that chronic Aβ application decreased the number of network bursts and spikes in a burst. The spatial structure of neural networks was also disturbed that characterized by reduction in both the number of key network elements (hubs) and connections between network elements. Moreover, application of brain-derived neurotrophic factor (BDNF) recombinant protein and BDNF hyperexpression by an adeno-associated virus vector partially prevented these amyloidopathy-induced neurodegenerative phenomena. BDNF maintained cell viability and spontaneous bioelectrical and calcium network activity in primary hippocampal cultures.

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Section: Discussionmentioning

confidence: 99%

Brain-Derived Neurotrophic Factor (BDNF) Preserves the Functional Integrity of Neural Networks in the β-Amyloidopathy Model in vitro

Митрошина

Yarkov

Mishchenko

et al. 2020

Front. Cell Dev. Biol.

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“…Cell treatments were performed in serum-free RPMI medium for 24 h. The stimuli used were: 100 ng/mL LPS (lipopolysaccharide from E. coli 0111:B4, Sigma-Aldrich, St. Louis, MO, USA); 25 μM paraquat (PQ; Sigma-Aldrich, St. Louis, MO, USA); 20 ng/mL IL-4 + 50 ng/mL IL-13 (PeproTech, Hamburg, Germany); and 1 μM Aβ oligomers (Bachem, Bubendorf, Switzerland). Aβ oligomers were prepared as described by Nuñez et al [ 39 ]. After treatments, cells were collected to analyze membranes and intracellular proteins, while extracellular media were cleaned from detached cells and debris (as indicated below) and stored at −80 °C until processed.…”

Section: Methodsmentioning

confidence: 99%

Evolutionary Origin of Insulin-Degrading Enzyme and Its Subcellular Localization and Secretion Mechanism: A Study in Microglial Cells

Corraliza-Gomez

Lillo

Cózar‐Castellano

et al. 2022

Cells

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The insulin-degrading enzyme (IDE) is a zinc-dependent metalloendopeptidase that belongs to the M16A metalloprotease family. IDE is markedly expressed in the brain, where it is particularly relevant due to its in vitro amyloid beta (Aβ)-degrading activity. The subcellular localization of IDE, a paramount aspect to understand how this enzyme can perform its proteolytic functions in vivo, remains highly controversial. In this work, we addressed IDE subcellular localization from an evolutionary perspective. Phylogenetic analyses based on protein sequence and gene and protein structure were performed. An in silico analysis of IDE signal peptide suggests an evolutionary shift in IDE exportation at the prokaryote/eukaryote divide. Subcellular localization experiments in microglia revealed that IDE is mostly cytosolic. Furthermore, IDE associates to membranes by their cytoplasmatic side and further partitions between raft and non-raft domains. When stimulated, microglia change into a secretory active state, produces numerous multivesicular bodies and IDE associates with their membranes. The subsequent inward budding of such membranes internalizes IDE in intraluminal vesicles, which later allows IDE to be exported outside the cells in small extracellular vesicles. We further demonstrate that such an IDE exportation mechanism is regulated by stimuli relevant for microglia in physiological conditions and upon aging and neurodegeneration.

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“…However, preparation of Aβ oligomers constitutes a complex method, and the difficulty to obtain reproducible oligomer preparations between different batches is the main reason for the existence of different protocols (Caballero et al, 2016). Based on the methods described by Núñez and coworkers (Núñez et al, 2018), we designed a new protocol to prepare Aβ oligomers, which is summarized in The tubes were spun in a SpeedVac centrifuge (800g, 10 minutes at RT, less than 25ºC to avoid peptide denaturalization) to remove the HFIP and to get the Aβ peptide as a film. The peptide film was then resuspended in 14.6 μl of DMSO to get Aβ 5 mM.…”

Section: Amyloid β 1-42 (Aβ1-42) Oligomers Preparationmentioning

confidence: 99%

The insulin-degrading enzyme: from molecular evolution and subcellular localization to new roles in microglial physiology

Gómez¹

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under our supervision, within the doctoral programme 1 DOCTORADO EN INVESTIGACIÓN BIOMÉDICA, under the modality of INTERNATIONAL DOCTOR MENTION, after taking into account the following considerations 2 : The candidate has fulfilled all the requirements and the level of training necessary to aspire to the Doctor's Degree with International Mention by the University of Valladolid, by performing an original research project. She has significantly contributed to the design of the scientific questions to be solved, framed at the interface of several disciplines, has designed, executed and analysed the experiments, and has performed data presentations in different formats (writing published work and presenting at scientific meetings), contributing to the dissemination of her results to the scientific community.

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Neurotoxic Ca2+ Signaling Induced by Amyloid–β Oligomers in Aged Hippocampal Neurons In Vitro

Cited by 5 publications

References 15 publications

Brain-Derived Neurotrophic Factor (BDNF) Preserves the Functional Integrity of Neural Networks in the β-Amyloidopathy Model in vitro

Brain-Derived Neurotrophic Factor (BDNF) Preserves the Functional Integrity of Neural Networks in the β-Amyloidopathy Model in vitro

Evolutionary Origin of Insulin-Degrading Enzyme and Its Subcellular Localization and Secretion Mechanism: A Study in Microglial Cells

The insulin-degrading enzyme: from molecular evolution and subcellular localization to new roles in microglial physiology

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