FISH analysis of aging-associated aneuploidy in neurons and nonneuronal brain cells

Grasiella A. Andriani, Cristina Montagna

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Aging is a ubiquitous complex process characterized by tissue degeneration and loss of cellular fitness. Genome instability (GIN) has long been implicated as a main causal factor in aging. The most severe form of genomic instability is whole chromosome instability (W-CIN), a state where dysfunction in chromosome segregation leads to whole chromosomes gains and losses. Aneuploidy is commonly linked to pathological states. It is a hallmark of spontaneous abortions and birth defects and it is observed virtually in every human tumor. There is mounting evidence that W-CIN increases with age, with the underlying hypothesis that some of the age-related loss of fitness phenotypes may be the result of W-CIN. Methodologically, the detection of stochastic W-CIN during the aging process poses unique challenges: aneuploid cells are scattered among diploid cells and, contrary to the cancer genome where aneuploidy is present in the background of massive ploidy changes, the number of aneuploid chromosome per cells is usually low (few per cell). Aging-associated aneuploidy is also largely stochastic or with limited clonal expansion. Therefore analysis at the single-cell level and the examination of a large number of cells is necessary. Here we describe a modification of the standard fluorescent in situ hybridization (FISH) protocol adapted for the detection of low-frequency mosaic aneuploidy in interphase cells isolated from the adult brain or within frozen tissue sections. This approach represents a straightforward method for the single-cell analysis of W-CIN in mammalian cells. It is based on the combination of four probes mapping to two different chromosomes and analysis of interphase cells, highly reducing false positives and enabling studying W-CIN also in post-mitotic tissues.

Original languageEnglish (US)
Title of host publicationNeuromethods
PublisherHumana Press Inc.
Pages271-298
Number of pages28
Volume131
DOIs
StatePublished - 2017

Publication series

NameNeuromethods
Volume131
ISSN (Print)0893-2336
ISSN (Electronic)1940-6045

Fingerprint

Chromosomal Instability
Aneuploidy
Chromosomes
Fluorescence In Situ Hybridization
Neurons
Brain
Aging of materials
Single-Cell Analysis
Genomic Instability
Interphase
Tissue
Chromosome Segregation
Ploidies
Frozen Sections
Spontaneous Abortion
Diploidy
Genes
Neoplasms
Cell Count
Genome

Keywords

  • Aging
  • Aneuploidy
  • Brain
  • FISH (Fluorescent in situ hybridization)
  • Fluorophores
  • Genomic instability (GIN)
  • Interphase FISH
  • NeuN+
  • NeuN−
  • Whole chromosome instability (W-CIN)

ASJC Scopus subject areas

  • Neuroscience(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Pharmacology, Toxicology and Pharmaceutics(all)
  • Psychiatry and Mental health

Cite this

Andriani, G. A., & Montagna, C. (2017). FISH analysis of aging-associated aneuploidy in neurons and nonneuronal brain cells. In Neuromethods (Vol. 131, pp. 271-298). (Neuromethods; Vol. 131). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-7280-7_14

FISH analysis of aging-associated aneuploidy in neurons and nonneuronal brain cells. / Andriani, Grasiella A.; Montagna, Cristina.

Neuromethods. Vol. 131 Humana Press Inc., 2017. p. 271-298 (Neuromethods; Vol. 131).

Research output: Chapter in Book/Report/Conference proceedingChapter

Andriani, GA & Montagna, C 2017, FISH analysis of aging-associated aneuploidy in neurons and nonneuronal brain cells. in Neuromethods. vol. 131, Neuromethods, vol. 131, Humana Press Inc., pp. 271-298. https://doi.org/10.1007/978-1-4939-7280-7_14
Andriani GA, Montagna C. FISH analysis of aging-associated aneuploidy in neurons and nonneuronal brain cells. In Neuromethods. Vol. 131. Humana Press Inc. 2017. p. 271-298. (Neuromethods). https://doi.org/10.1007/978-1-4939-7280-7_14
Andriani, Grasiella A. ; Montagna, Cristina. / FISH analysis of aging-associated aneuploidy in neurons and nonneuronal brain cells. Neuromethods. Vol. 131 Humana Press Inc., 2017. pp. 271-298 (Neuromethods).
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