Transcriptomics and Live Imaging to Define Functional Phenotypes of Microglia in Pathological Human Tissue

Area of Science:

• Neuroscience
• Immunology
• Cell Biology

Background:

• Microglia, the resident immune cells of the central nervous system, are increasingly recognized for their diverse functional states.
• Understanding human microglial phenotypes is crucial for comprehending brain health and disease, yet direct analysis is challenging.

Purpose of the Study:

• To establish protocols for analyzing human microglial phenotypes from surgically resected brain tissue.
• To investigate microglial characteristics, including morphology, marker expression, motility, and transcriptional profiles.
• To examine how microglial transcriptomes change following a seizure event in epilepsy patients.

Main Methods:

• Utilized therapeutic excision of brain tissue from patients with temporal lobe epilepsies and cortical gliomas.
• Developed techniques for microglial identification, morphological assessment, and state-specific marker expression analysis.
• Measured resting and induced microglial motility and performed transcriptome analysis to define gene expression profiles.

Main Results:

• Successfully identified and characterized human microglia from patient brain samples.
• Established methods to assess microglial shape, marker expression, and motility.
• Defined a baseline human microglial transcriptome and observed alterations post-seizure.

Conclusions:

• The developed protocols enable detailed characterization of human microglial phenotypes in vivo.
• Seizures induce significant changes in the human microglial transcriptome.
• This work provides a foundation for studying dynamic microglial responses in neurological disorders.