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We studied effective connectivity in rat cortical cultures with various degrees of spatial aggregation, ranging from homogeneous networks to highly aggregated ones. We considered small cultures 3 mm in diameter and that contained about 2;000 neurons. Spatial inhomogeneity favored an increase of metric correlations and connectivity among neighboring neurons. Effective connectivity was determined from spontaneous activity recordings using calcium fluorescence imaging. We used generalized transfer entropy as tool to infer the effective connectivity. We carried out numerical simulations to build networks that mimicked the experimental ones and to test the reliability of the connectivity-inference algorithm. Effective connectivity traits were investigated during the development of the cultures over two weeks, and along the gradual blockade of excitatory connections through CNQX. We observed that the average effective connectivity rapidly increased during culture development. At day in vitro (DIV) 15 the average excitatory in-degree was measured as &#x2243;kinE 50 for homogeneous and semi aggregated networks, and &#x2243;kinE 120 for aggregated ones, and with 20 percent inhibition. Aggregated cultures exhibited assortative traits and a high resilience to chemical damage, while the other cultures were dissassortative or neutral, and less resilient. Our work illustrates the role of metric correlations in spatially embedded networks in shaping connectivity and activity traits in living neuronal networks.

Neuronal Spatial Arrangement Shapes Effective Connectivity Traits of in vitro Cortical Networks