Visualization and Quantification of Post-stroke Neural Connectivity and Neuroinflammation Using Serial Two-Photon...

Area of Science:

• Neuroscience
• Immunology
• Medical Imaging

Background:

• Stroke induces neuroinflammation and neuroplasticity, impacting recovery.
• Understanding these changes across the whole brain is crucial for effective treatment.

Purpose of the Study:

• To develop and validate a whole-brain image analysis pipeline for quantifying neuroinflammation and neuroplasticity post-stroke.
• To investigate long-term motor circuit connectivity and sub-acute neuroinflammation in mouse stroke models.

Main Methods:

• Serial two-photon tomography (STPT) for high-resolution whole-brain imaging.
• Supervised machine learning (random forest models) for automated cellular signal detection.
• Registration to a standardized 3-D mouse brain atlas (Common Coordinate Framework v3.0).
• Viral tracing (pseudorabies virus) to map neural connectivity.
• Intravenous injection of labeled immune cells (CD8+ T cells) to study neuroinflammation.

Main Results:

• The pipeline successfully detected and quantified fluorescent signals, enabling unbiased analysis of cellular changes.
• Region-specific quantification of neural connectivity and neuroinflammation was achieved.
• The study demonstrated the utility of the workflow in evaluating neuronal and lymphocyte populations in healthy and injured brains.

Conclusions:

• The developed STPT-based workflow provides a robust framework for objective evaluation of neuroinflammation and neuroplasticity post-stroke.
• This method can be a critical tool for investigating stroke recovery mechanisms and other brain injuries or diseases.
• The findings highlight the role of remote brain regions in stroke pathology and recovery.