Molecular Biology of Neuronal Cells

NeuroProof integrates advanced MEA-neurochip technology with cutting-edge molecular biology assays and bioinformatics tools to enhance, optimize, and validate experimental outcomes. This approach allows us to correlate functional data with molecular insights.

Our analytical toolkit for validation includes quantitative assessments of tissue-specific cell composition (neurons, glial cells), alongside receptor detection and status. These analyses run in parallel with functional compound screening on MEA platforms, supporting developmental and stem cell assays. This enables real-time monitoring of compound effects at the cellular and molecular levels.

Key Techniques:

• Western Blotting: Utilized for the identification and quantification of specific proteins crucial for validation.

• Imaging: We employ automated confocal microscopy for detailed imaging studies.

• AAV Gene Constructs: Our optimized adeno-associated virus constructs facilitate genetic manipulation within neuronal cell cultures, demonstrated by successful transductions such as alpha-synuclein overexpression or SCN1A gene knockdown to simulate disease models.

• Transcriptomics: RNASeq analysis characterizes cellular gene expression profiles, offering insights into target expression or compound effects.

• Energy Metabolism: Crucial for assessing CNS metabolomic networks, our energy metabolism analysis employs transcriptomic and proteomic data. Using systems biology modeling, we simulate substrate concentrations to quantify dysfunctions in energy metabolism, including key processes like ATP production and fatty acid metabolism. 

These data support the MEA analyses especially in our developmental assays and stem cell assays in a way that compound-dependent effects can be monitored at the level of associated cell types and receptors.

Image: Developmental stage-dependent concentration/response experiments show increasing potency which is in agreement with receptor expression