This work presents an analytical pipeline for inferring network-level properties of in vitro cortical cultures from multichannel electrophysiological recordings and proves the validity of the RC-based connectivity inference and establishes a scalable, data-driven framework for functional network characterization in neuronal culture systems.
Abstract
Graph-theoretical analysis offers a principled framework for quantifying emergent dynamics in neuronal cultures. Here, we present an analytical pipeline for inferring network-level properties of in vitro cortical cultures from multichannel electrophysiological recordings. The approach builds on a recently proposed Reservoir Computing (RC) framework (Auslender et al., 2025), which enables direct extraction of an Intrinsic Connectivity Map (ICM) from neural activity. We interpret the ICM as an effective adjacency matrix and apply graph-theoretic centrality measures to quantify node- and edge-level contributions to the culture's collective dynamics. We systematically evaluate both local and global graph metrics and examine their relationships with experimentally measured activity features, including firing rates and network-level descriptors. To validate the inference procedure, we also simulate the experimental environment, enabling controlled benchmarking of the RC-derived connectivity against a known ground-truth adjacency matrix and assessment of model performance as a function of graph structure. Our results demonstrate statistically robust associations, of varying strength, between graph-theoretic measures and experimentally observed activity patterns. These findings additionally support the validity of the RC-based connectivity inference and establish a scalable, data-driven framework for functional network characterization in neuronal culture systems.
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