Analysis of the Microglial Sensome
Suzanne E Hickman1, Joseph El Khoury2
1Center for Immunology and Inflammatory Diseases, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA. Shickman@mgh.harvard.edu.
Methods in Molecular Biology (Clifton, N.J.)
|August 9, 2019
Summary
Researchers developed methods to isolate microglia from mouse brains and analyze their "sensome" genes and proteins. This helps study microglial sensing functions in health and disease.
Area of Science:
- Neuroscience
- Immunology
- Cell Biology
Background:
- Microglia are crucial brain immune cells responsible for sensing environmental changes.
- Microglial sensing functions are regulated by a specific set of genes known as the sensome.
- Assessing sensome gene and protein expression in isolated microglia is vital for understanding their roles.
Purpose of the Study:
- To provide a detailed protocol for isolating microglia from adult mouse brains.
- To demonstrate methods for analyzing sensome gene and protein expression in isolated microglia.
- To enable the study of sensome expression in various physiological and pathological conditions.
Main Methods:
- Isolation of microglia from adult mouse brains.
- Quantitative PCR (qPCR) for analyzing sensome gene expression.
- Flow cytometry for analyzing sensome protein expression ex vivo.
- In situ hybridization for detecting sensome RNA within the mouse brain.
Main Results:
- A step-by-step procedure for microglia isolation was successfully established.
- Examples of analyzing sensome gene and protein expression using qPCR and flow cytometry were provided.
- A method for in situ hybridization to detect sensome RNA in brain tissue was described.
Conclusions:
- The described methods allow for the assessment of sensome expression in isolated microglia.
- These techniques are applicable to all known sensome genes.
- The developed approaches will aid in investigating microglial sensing in both normal and diseased states.
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