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Dissection of the Drosophila Pupal Retina for Immunohistochemistry, Western Analysis, and RNA Isolation
Published on: March 15, 2019
Genome-wide analyses of steroid- and radiation-triggered programmed cell death in Drosophila
Cheng-Yu Lee1, Emily A Clough, Paula Yellon
1Center for Biosystems Research, University of Maryland Biotechnology Institute and Department of Medicine, University of Maryland, College Park, MD 20742, USA.
Abstract:
Apoptosis and autophagy are two forms of programmed cell death that play important roles in the removal of unneeded and abnormal cells during animal development. While these two forms of programmed cell death are morphologically distinct, recent studies indicate that apoptotic and autophagic cell death utilize some common regulatory mechanisms. To identify genes that are associated with apoptotic and autophagic cell death, we monitored changes in gene transcription by using microarrays representing nearly the entire Drosophila genome. Analyses of steroid-triggered autophagic cell death identified 932 gene transcripts that changed 5-fold or greater in RNA level. In contrast, radiation-activated apoptosis resulted in 34 gene transcripts that exhibited a similar magnitude of change. Analyses of these data enabled us to identify genes that are common and unique to steroid- and radiation-induced cell death. Mutants that prevent autophagic cell death exhibit altered levels of gene transcription, including genes encoding caspases, non-caspase proteases, and proteins that are similar to yeast autophagy proteins. This study also identifies numerous novel genes as candidate cell death regulators and suggests new links between apoptosis and autophagic cell death.
Insights
This study investigated programmed cell death in Drosophila, comparing apoptosis and autophagy. Researchers identified common and unique genes involved in both processes, revealing new regulatory links.
Area of Science:
- Cell Biology
- Genetics
- Developmental Biology
Background:
- Programmed cell death, including apoptosis and autophagy, is crucial for animal development.
- While morphologically distinct, apoptosis and autophagy share regulatory mechanisms.
- Understanding these shared pathways is key to identifying novel cell death regulators.
Purpose of the Study:
- To identify genes associated with apoptotic and autophagic cell death.
- To compare gene transcription changes in response to different cell death triggers.
- To uncover novel genes and regulatory links between apoptosis and autophagy.
Main Methods:
- Utilized microarrays to monitor genome-wide gene transcription changes in Drosophila.
- Analyzed gene expression profiles following steroid-induced autophagic cell death.
- Examined gene expression changes during radiation-activated apoptosis.
Main Results:
- Steroid-induced autophagy affected 932 gene transcripts (≥5-fold change).
- Radiation-induced apoptosis affected 34 gene transcripts (≥5-fold change).
- Identified common and unique genes for both cell death types, including caspase and autophagy-related genes.
Conclusions:
- Discovered numerous novel candidate genes regulating cell death.
- Highlighted shared and distinct genetic pathways in apoptosis and autophagy.
- Suggested new molecular links connecting apoptosis and autophagic cell death.
