Effects of Cerebrospinal Fluids from Alzheimer and Non-Alzheimer Patients on Neurons-Astrocytes-Microglia Co-Culture

Int J Mol Sci. 2024 Feb 21;25(5):2510. doi: 10.3390/ijms25052510.

Abstract

Alzheimer's disease (AD) is the most common form of dementia, characterized by the accumulation of β-amyloid plaques, tau tangles, neuroinflammation, and synaptic/neuronal loss, the latter being the strongest correlating factor with memory and cognitive impairment. Through an in vitro study on a neurons-astrocytes-microglia (NAM) co-culture system, we analyzed the effects of cerebrospinal fluid (CSF) samples from AD and non-AD patients (other neurodegenerative pathologies). Treatment with CSF from AD patients showed a loss of neurofilaments and spheroids, suggesting the presence of elements including CX3CL1 (soluble form), destabilizing the neurofilaments, cellular adhesion processes, and intercellular contacts. The NAM co-cultures were analyzed in immunofluorescence assays for several markers related to AD, such as through zymography, where the expression of proteolytic enzymes was quantified both in cell extracts and the co-cultures' conditioned medium (CM). Through qRT-PCR assays, several genes involved in the formation of β-amyloid plaque, in phosphorylation of tau, and in inflammation pathways and MMP expression were investigated.

Keywords: Alzheimer; CX3CL1; neuroinflammation; tau; β-amyloid.

MeSH terms

  • Alzheimer Disease* / metabolism
  • Amyloid beta-Peptides / metabolism
  • Astrocytes / metabolism
  • Coculture Techniques
  • Humans
  • Microglia / metabolism
  • Neurons / metabolism
  • tau Proteins / metabolism

Substances

  • tau Proteins
  • Amyloid beta-Peptides

Grants and funding

This research was funded by the University of Palermo (www.UniPa.it (accessed on 1 June 2010)), fund FFR 2022 Ghersi and fund R4D15-P8MSRI07_MARGINE (Ghersi).