Automated Immunofluorescence Staining for Analysis of DNA Damage and Apoptosis in Brain Sections

Gabriela De la Cruz; Nana Nikolaishvili-Feinberg; Timothy R Gershon

doi:10.1007/978-1-0716-2752-5_6

Automated Immunofluorescence Staining for Analysis of DNA Damage and Apoptosis in Brain Sections

Methods Mol Biol. 2023:2583:55-61. doi: 10.1007/978-1-0716-2752-5_6.

Authors

Gabriela De la Cruz^{1

2}, Nana Nikolaishvili-Feinberg^{1

2}, Timothy R Gershon^{3

4

5}

Affiliations

¹ Lineberger Comprehensive Cancer Center, The University of North Carolina at Chapel Hill, Chapel Hill, NC, USA.
² Pathology Services Core, The University of North Carolina at Chapel Hill, Chapel Hill, NC, USA.
³ Lineberger Comprehensive Cancer Center, The University of North Carolina at Chapel Hill, Chapel Hill, NC, USA. gershont@neurology.unc.edu.
⁴ Department of Neurology, The University of North Carolina at Chapel Hill, Chapel Hill, NC, USA. gershont@neurology.unc.edu.
⁵ UNC Neuroscience Center, The University of North Carolina at Chapel Hill, Chapel Hill, NC, USA. gershont@neurology.unc.edu.

PMID: 36418725
DOI: 10.1007/978-1-0716-2752-5_6

Abstract

Neural progenitors show a strong tendency to undergo apoptosis in response to DNA damage, and both impaired DNA repair and increased neural progenitor apoptosis are associated with microcephaly. Here we present an immunohistochemistry-based method for assessing DNA damage and apoptosis in the neonatal mouse brain. These methods are suitable for determining in specific experimental conditions the fractions of cells with DNA double-strand breaks, the fractions of cells undergoing apoptosis, or both. While DNA damage in neural progenitors can trigger apoptosis, inappropriate apoptosis may also result from other processes. Simultaneous analysis of DNA damage and apoptosis in mouse models of microcephaly can determine how genetic instability and cell death contribute to the observed phenotype.

Keywords: Apoptosis; DNA damage; Microcephaly; Neural progenitors.

MeSH terms

Animals
Apoptosis
Brain
DNA Damage
Fluorescent Antibody Technique
Mice
Microcephaly*
Staining and Labeling