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Analysis of Translation in the Developing Mouse Brain using Polysome Profiling
Published on: May 22, 2021
Characterization of polysomes purified from human postmortem brain tissues.
N S Långström1, A Eriksson, B G Winblad
1Department of Psychiatry and Fishberg Center for Neurobiology, Mount Sinai School of Medicine, New York, N.Y., U.S.A.; Department of Geriatric Medicine, Karolinska Institute, Huddinge University Hospital, S-14186 Huddinge, Sweden.
Neurochemistry International
|May 28, 2010
Summary
Researchers successfully isolated intact, translationally active cytosolic polyribosomes (polysomes) from human postmortem brain tissue. This method enables studies on translational regulation of gene expression in the human brain under various conditions.
Area of Science:
- Neuroscience
- Molecular Biology
- Genetics
Background:
- Translational regulation of gene expression is crucial in the human brain.
- Investigating this process requires robust methods for isolating active cytosolic polyribosomes (polysomes).
- Human postmortem brain tissue presents challenges due to variability in age, postmortem interval, and agonal state.
Purpose of the Study:
- To evaluate the suitability of human postmortem brain tissue for studying translational regulation.
- To establish a reproducible method for purifying cytosolic polyribosomes (polysomes) from human brain.
- To compare human polysomes with those from rat brains.
Main Methods:
- Cytosolic polyribosomes (polysomes) were purified from human postmortem frontal cortex and cerebellum.
- Purified polysomes were analyzed for yield, size, and translational activity.
- Human polysomes were compared to polysomes isolated from rat brains.
Main Results:
- Similar yields of polysomes were obtained from human frontal cortex, irrespective of age, postmortem interval, or agonal state.
- Human polysomes contained fewer ribosomes per mRNA (up to 6) compared to rat polysomes (15-20).
- Human polysomes exhibited lower translational efficiency than rat polysomes, but elongation rates were similar.
Conclusions:
- Intact and translationally active polysomes can be reproducibly isolated from human postmortem brain tissues.
- The described methodology is suitable for investigating translational control of gene expression in the human brain.
- These polysomes can be utilized in studies of both normal and neuropathological conditions.
