Monitoring Cellular Proliferation and Apoptosis in Atherosclerosis Plaques and Intimal Thickenings

Research output: Chapter in Book/Report/Conference proceedingChapter in a book

Abstract

Immunohistochemistry for specific proteins characteristic of proliferative or apoptotic cells allows for monitoring of these cell behaviors in biological tissues samples, including atherosclerotic plaques and intimal thickenings. Proliferating cell nuclear antigen (PCNA) and Ki-67 are widely used markers of cell proliferation and cleaved caspase-3 is a well-established marker of apoptosis that can be detected in tissue samples using immunohistochemistry. This technique enables quantification of the abundance of these proteins and provides information on the distribution of these biomarkers in tissues. By combining with immunohistochemistry for specific cell type markers, it is also possible to determine which cell types are proliferating or undergoing apoptosis. Here, we detail protocols for immunohistochemistry of PCNA, Ki-67, and cleaved caspase-3 for evaluation of cellular proliferation and apoptosis in atherosclerotic plaques in vivo. In addition, we outline methods for the quantification and localization of cell proliferation using bromodeoxyuridine/5-bromo-2'-deoxyuridine (BrdU) and ethynyldeoxyuridine/5-ethynyl-2 ́-deoxyuridine(EdU) labeled tissue samples collected from animals exposed to BrdU or EdU.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
Pages507-519
Number of pages13
Volume2419
DOIs
Publication statusPublished - 1 Apr 2022

Publication series

NameMethods in Molecular Biology
Volume2419
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Bibliographical note

Funding Information:
Jason Johnson is funded by British Heart Foundation Senior Research Fellowship, FS/18/1/33234.

Publisher Copyright:
© 2022, The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.

Keywords

  • Animals
  • Apoptosis
  • Atherosclerosis
  • Bromodeoxyuridine/metabolism
  • Cell Division
  • Cell Proliferation
  • Ki-67 Antigen/metabolism
  • Plaque, Atherosclerotic
  • Proliferating Cell Nuclear Antigen/metabolism

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