Free-floating adult human brain-derived slice cultures as a model to study the neuronal impact of Alzheimer's disease-associated Aβ oligomers.

Autor: Mendes ND; Dept. Biochemistry and Immunology, Ribeirao Preto Medical School, University of Sao Paulo, SP, Brazil; Dept. Pathology and Forensic Medicine, Ribeirao Preto Medical School, University of Sao Paulo, SP, Brazil., Fernandes A; Dept. Biochemistry and Immunology, Ribeirao Preto Medical School, University of Sao Paulo, SP, Brazil; Dept. Physiology, Ribeirão Preto Medical School, University of São Paulo, SP, Brazil., Almeida GM; Dept. Biochemistry and Immunology, Ribeirao Preto Medical School, University of Sao Paulo, SP, Brazil., Santos LE; Institute of Medical Biochemistry, Federal University of Rio de Janeiro, RJ, Brazil., Selles MC; Institute of Medical Biochemistry, Federal University of Rio de Janeiro, RJ, Brazil., Lyra E Silva NM; Institute of Medical Biochemistry, Federal University of Rio de Janeiro, RJ, Brazil., Machado CM; Department of Anatomy, Institute of Biosciences, São Paulo State University, SP, Brazil., Horta-Júnior JAC; Department of Anatomy, Institute of Biosciences, São Paulo State University, SP, Brazil., Louzada PR; Institute of Biomedical Sciences, Federal University of Rio de Janeiro, RJ, Brazil., De Felice FG; Institute of Medical Biochemistry, Federal University of Rio de Janeiro, RJ, Brazil; Centre for Neuroscience Studies, Department of Biomedical and Molecular Sciences, Queen's University, Kingston, ON, Canada., Alves-Leon S; Hospital Universitário Clementino Fraga Filho, Federal University of Rio de Janeiro, RJ, Brazil., Marcondes J; Hospital Universitário Clementino Fraga Filho, Federal University of Rio de Janeiro, RJ, Brazil., Assirati JA Jr; Ribeirão Preto Medical School Clinical Hospital, University of São Paulo, SP, Brazil., Matias CM; Ribeirão Preto Medical School Clinical Hospital, University of São Paulo, SP, Brazil., Klein WL; Department of Neurobiology, Northwestern University, IL, USA., Garcia-Cairasco N; Dept. Physiology, Ribeirão Preto Medical School, University of São Paulo, SP, Brazil., Ferreira ST; Institute of Medical Biochemistry, Federal University of Rio de Janeiro, RJ, Brazil; Institute of Biophysics Carlos Chagas Filho, Federal University of Rio de Janeiro, RJ, Brazil., Neder L; Dept. Pathology and Forensic Medicine, Ribeirao Preto Medical School, University of Sao Paulo, SP, Brazil; Barretos Cancer Hospital, Barretos, SP, Brazil., Sebollela A; Dept. Biochemistry and Immunology, Ribeirao Preto Medical School, University of Sao Paulo, SP, Brazil. Electronic address: sebollela@fmrp.usp.br.
Jazyk: angličtina
Zdroj: Journal of neuroscience methods [J Neurosci Methods] 2018 Sep 01; Vol. 307, pp. 203-209. Date of Electronic Publication: 2018 May 31.
DOI: 10.1016/j.jneumeth.2018.05.021
Abstrakt: Background: Slice cultures have been prepared from several organs. With respect to the brain, advantages of slice cultures over dissociated cell cultures include maintenance of the cytoarchitecture and neuronal connectivity. Slice cultures from adult human brain have been reported and constitute a promising method to study neurological diseases. Despite this potential, few studies have characterized in detail cell survival and function along time in short-term, free-floating cultures.
New Method: We used tissue from adult human brain cortex from patients undergoing temporal lobectomy to prepare 200 μm-thick slices. Along the period in culture, we evaluated neuronal survival, histological modifications, and neurotransmitter release. The toxicity of Alzheimer's-associated Aβ oligomers (AβOs) to cultured slices was also analyzed.
Results: Neurons in human brain slices remain viable and neurochemically active for at least four days in vitro, which allowed detection of binding of AβOs. We further found that slices exposed to AβOs presented elevated levels of hyperphosphorylated Tau, a hallmark of Alzheimer's disease.
Comparison With Existing Method(s): Although slice cultures from adult human brain have been previously prepared, this is the first report to analyze cell viability and neuronal activity in short-term free-floating cultures as a function of days in vitro.
Conclusions: Once surgical tissue is available, the current protocol is easy to perform and produces functional slices from adult human brain. These slice cultures may represent a preferred model for translational studies of neurodegenerative disorders when long term culturing in not required, as in investigations on AβO neurotoxicity.
(Copyright © 2018 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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