Background & Aims
The regulation of chronic pain is influenced by the neuroinflammatory response of microglia. Microglia play a crucial role in maintaining brain homeostasis, but when over-activated or uncontrolled, they can contribute to various brain diseases, promoting neuronal cell death in neurodegenerative conditions like Alzheimer’s, Parkinson’s, Huntington’s, and ALS [1]. Similarly, recent advancements indicate that the extracellular matrix undergoes remodeling in neurodegenerative diseases. This remodeling influences the release of pro- and anti-inflammatory cytokines, shaping the fate of immune, glial, and neuronal cells [2]. These comprehensive insights fuel our interest in understanding microglial responses to matrix rigidity and contribute valuable knowledge to the broader field of neurobiology. This research aims to investigate how matrix rigidity influences microglial behavior, function, and epigenetic programming.
Methods
To investigate the role of substrate biomechanics on BV-2 cell morphology and function in different matrix rigidities of 0.2 kPa, 0.5 kPa, 2.0 kPa, 8.0 kPa, 16.0 kPa, 32.0 kPa, and 64.0 kPa. The cells are cultured in F-12 HAMS, FBS, and STREP media in 6-well plates coated with various extracellular matrix conditions.
1.Morphology: Microglial morphology will be analyzed in terms of area, circularity, and process complexity.
2.Engulfment: Functional assessments will be performed by placing carboxyl beads into the cultured microglia in 6-well ECM-coated plates. After incubation, the dynamics of glial engagement with the carboxyl beads will be recorded and analyzed.
3.Epigenetic Programming: the result of microglia’s engagement with the ECM will be studied using DNA methylation patterns, histone modifications, and gene expression analysis.
4.Microscopic videos will be recorded to generate predictive computational models for microglial behavior.
Results
Our preliminary results suggest that substrate biomechanics affects the branching of BV-2 processes as well as cellularity and engulfment(function). These analyses are currently being completed. We are currently working on researching changes in DNA methylation patterns, histone modifications, and gene expression.
Conclusions
This study aims to understand the role of the extracellular matrix in glial behavior and function. The results can open avenues for biomarker discovery. Such biomarkers could serve as diagnostic tools for early detection and intervention in diseases like Alzheimer’s and Parkinson’s. In addition, understanding the mechanics of the extracellular matrix is a new therapeutic approach. Instead of targeting microglia, we can explore treatments that alter how the extracellular matrix behaves, to develop potential approaches to treat neurodegenerative diseases.
References
1.Cheray, M., & Joseph, B. (2018). Epigenetics control microglia plasticity. Frontiers in cellular neuroscience, 12, 243.
2.Wiemann, S., Reinhard, J., & Faissner, A. (2019). Immunomodulatory role of the extracellular matrix protein tenascin-C in neuroinflammation. Biochemical Society Transactions, 47(6).
Presenting Author
Vaneeza Kausar
Poster Authors
Topics
- Models: Transition to Chronic Pain