The chemical substances of synthetic origin and foreign to the biological systems are known as xenobiotics, such as drugs, medicines, pesticides, fertilizers, cosmetics, preservatives, food additives, industrial chemicals, and other environmental pollutants generated either through anthropogenic or geological activities. The xenobiotics may adversely affect human life and result in various disease conditions. Apoptosis and autophagy are two well-regulated evolutionarily conserved self-destructive processes that maintain cellular homeostasis and cell fate. Apoptosis, also known as programmed cell death, is triggered by various stimuli, including xenobiotics. Cellular characteristics of apoptosis include chromatin condensation, cytoplasm shrinkage, and asymmetry in the plasma membrane. Autophagy involves massive accumulation of double-membrane vesicles (autophagosomes) containing damaged or waste cytoplasmic materials, which fuses with lysosomes and results in bulk degradation of the cytosolic components and thus is also considered type II cell death. Since both types of cell death mechanisms are expressed in several cell systems, including the immunological, neurological, hepatic, intestinal, cutaneous, and reproductive, it is appealing as a generic predictor of response. Depending on the stimulus and kind of cell, an apoptotic cell exhibits a wide range of distinctive morphological and biochemical characteristics. Flow and imaging cytometry allows for the quick examination of the vast majority of traditional apoptotic markers. Thus, cytometry emerged as the preferred tool for various cellular death investigations. The last two decades have seen the development of a wide range of cytometric techniques intended to detect apoptotic cells and investigate the causes behind this type of cell death. Flowcytometric analysis has been used for many years to detect the stages of apoptosis and autophagy. This chapter aims to provide curated information on applying flowcytometric methods to detect different stages of apoptosis and autophagy, including using pivotal assays, which can detect both early and late apoptosis. Further, the chapter also deals with a description of different dyes or stains, such as monodansylcadaverine, acridine orange, LysoTracker, and autophagy-specific markers, like anti-sequestosome and anti-LC3-II, which are pretty helpful in staining or targeting the autophagic contents or autophagic vacuoles for detection and quantification of autophagic activity in response to different xenobiotic compounds.

Environmental Stressors-Mediated Apoptosis & Autophagy Detected Through Flow Cytometry

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发表日期：

2025年

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Meenu Srivastava,
AB Pant

摘要

The chemical substances of synthetic origin and foreign to the biological systems are known as xenobiotics, such as drugs, medicines, pesticides, fertilizers, cosmetics, preservatives, food additives, industrial chemicals, and other environmental pollutants generated either through anthropogenic or geological activities. The xenobiotics may adversely affect human life and result in various disease conditions. Apoptosis and autophagy are two well-regulated evolutionarily conserved self-destructive processes that maintain cellular homeostasis and cell fate. Apoptosis, also known as programmed cell death, is triggered by various stimuli, including xenobiotics. Cellular characteristics of apoptosis include chromatin condensation, cytoplasm shrinkage, and asymmetry in the plasma membrane. Autophagy involves massive accumulation of double-membrane vesicles (autophagosomes) containing damaged or waste cytoplasmic materials, which fuses with lysosomes and results in bulk degradation of the cytosolic components and thus is also considered type II cell death. Since both types of cell death mechanisms are expressed in several cell systems, including the immunological, neurological, hepatic, intestinal, cutaneous, and reproductive, it is appealing as a generic predictor of response. Depending on the stimulus and kind of cell, an apoptotic cell exhibits a wide range of distinctive morphological and biochemical characteristics. Flow and imaging cytometry allows for the quick examination of the vast majority of traditional apoptotic markers. Thus, cytometry emerged as the preferred tool for various cellular death investigations. The last two decades have seen the development of a wide range of cytometric techniques intended to detect apoptotic cells and investigate the causes behind this type of cell death. Flowcytometric analysis has been used for many years to detect the stages of apoptosis and autophagy. This chapter aims to provide curated information on applying flowcytometric methods to detect different stages of apoptosis and autophagy, including using pivotal assays, which can detect both early and late apoptosis. Further, the chapter also deals with a description of different dyes or stains, such as monodansylcadaverine, acridine orange, LysoTracker, and autophagy-specific markers, like anti-sequestosome and anti-LC3-II, which are pretty helpful in staining or targeting the autophagic contents or autophagic vacuoles for detection and quantification of autophagic activity in response to different xenobiotic compounds.