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Published on: August 29, 2018
Texture analysis of poly-adenylated mRNA staining following global brain ischemia and reperfusion
Jeffrey J Szymanski1, Jill T Jamison, Donald J DeGracia
1Department of Physiology, Wayne State University School of Medicine, Detroit, MI 48201, USA.
Abstract:
Texture analysis provides a means to quantify complex changes in microscope images. We previously showed that cytoplasmic poly-adenylated mRNAs form mRNA granules in post-ischemic neurons and that these granules correlated with protein synthesis inhibition and hence cell death. Here we utilized the texture analysis software MaZda to quantify mRNA granules in photomicrographs of the pyramidal cell layer of rat hippocampal region CA3 around 1h of reperfusion after 10min of normothermic global cerebral ischemia. At 1h reperfusion, we observed variations in the texture of mRNA granules amongst samples that were readily quantified by texture analysis. Individual sample variation was consistent with the interpretation that animal-to-animal variations in mRNA granules reflected the time-course of mRNA granule formation. We also used texture analysis to quantify the effect of cycloheximide, given either before or after brain ischemia, on mRNA granules. If administered before ischemia, cycloheximide inhibited mRNA granule formation, but if administered after ischemia did not prevent mRNA granulation, indicating mRNA granule formation is dependent on dissociation of polysomes. We conclude that texture analysis is an effective means for quantifying the complex morphological changes induced in neurons by brain ischemia and reperfusion.
Insights
Texture analysis quantifies mRNA granules in neurons after brain ischemia. This method reveals variations in granule formation, aiding understanding of neuronal response to ischemia and reperfusion.
Area of Science:
- Neuroscience
- Cell Biology
- Biophysics
Background:
- Cytoplasmic poly-adenylated mRNAs form granules in post-ischemic neurons.
- These mRNA granules correlate with protein synthesis inhibition and neuronal cell death.
- Quantifying these complex morphological changes is crucial for understanding neuronal injury.
Purpose of the Study:
- To utilize texture analysis software (MaZda) for quantifying mRNA granules in rat hippocampal neurons post-ischemia.
- To investigate variations in mRNA granule formation and their correlation with the time-course of neuronal response.
- To assess the impact of cycloheximide on mRNA granule formation before and after ischemia.
Main Methods:
- Texture analysis of photomicrographs from the pyramidal cell layer of rat hippocampal region CA3.
- Quantification of mRNA granules at 1h reperfusion after 10min global cerebral ischemia.
- Application of cycloheximide before and after ischemic events.
Main Results:
- Texture analysis effectively quantified variations in mRNA granule texture among samples.
- Animal-to-animal variations in mRNA granules correlated with the time-course of granule formation.
- Cycloheximide administered before ischemia inhibited granule formation, while post-ischemia administration did not, suggesting dependence on polysome dissociation.
Conclusions:
- Texture analysis is an effective tool for quantifying complex morphological changes in neurons induced by brain ischemia and reperfusion.
- mRNA granule formation is dependent on the dissociation of polysomes.
- Understanding these mechanisms can inform therapeutic strategies for ischemic brain injury.
