Raman microscopy at the subcellular level: a study on early apoptosis in endothelial cells induced by Fas ligand and cycloheximide

The Analyst
Krzysztof CzamaraAgnieszka Kaczor

Abstract

High spatially resolved Raman microscopy was applied to study the early apoptosis in endothelial cells and chemical and structural changes induced by this process. Application of cluster analysis enabled separation of signals due to various subcellular organelles and compartments such as the nuclei, nucleoli, endoplasmic reticulum or cytoplasm and analysis of alterations locally at the subcellular level. Different stimuli, i.e. Fas ligand, a tumor necrosis factor, and cycloheximide, an inhibitor of eukaryotic protein biosynthesis, were applied to induce apoptotic mechanisms. Due to different mechanisms of action, the changes observed in subcellular structures were different for FasL and cycloheximide. Although in both cases a statistically significant decrease of the protein level was observed in all studied cellular structures, the increase of the nucleic acids content locally in apoptotic nuclei was considerably more pronounced upon FasL-induced apoptosis compared to the cycloheximide one. Additionally, apoptosis invokes also a decrease of the proteins with the α-helix protein structure selectively for FasL in the cytoplasm and endoplasmic reticulum.

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Citations

Aug 23, 2017·Applied Spectroscopy·Faris SinjabIoan Notingher
Oct 9, 2018·Journal of Biophotonics·Krzysztof CzamaraAgnieszka Kaczor
Jun 15, 2019·The Analyst·Imesha W De SilvaGuido F Verbeck
Jun 15, 2020·Biochimica Et Biophysica Acta. Molecular and Cell Biology of Lipids·Marta Z PaciaMalgorzata Baranska
Apr 7, 2021·Journal of Medicinal Chemistry·Adriana AdamczykMalgorzata Baranska

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Apoptosis

Apoptosis is a specific process that leads to programmed cell death through the activation of an evolutionary conserved intracellular pathway leading to pathognomic cellular changes distinct from cellular necrosis