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Vincenzo Costanzo1, Ludovica Altieri1,2, Patrizia Lavia1 ( (1) Institute of Molecular Biology and Pathology (IBPM), CNR National Research Council, Rome, Italy; (2) Department of Biology and Biotechnology “Charles Darwin”, Sapienza University of Rome, Italy)
Centromere protein-F (CENP-F) is a large multidomain protein with well-characterized functions in regulating microtubule dynamics and chromosome segregation during mitosis. Mutations of CENP-F identified in microcephaly and ciliopathies suggest a critical role of this protein during brain development, which however remains poorly elucidated. Here, we have investigated CENP-F roles in cellular models of neuronal differentiation. First, we characterised morphological changes and appearance of neuronal markers in murine P19 pluripotent cells induced to differentiate in the presence of retinoic acid. We next extended these studies to dedifferentiated, neuroblastoma SKNBE(2) cells induced to re-establish a differentiated neuronal phenotype. We also performed CENP-F interference assays in SKNBE(2) cells to assess whether the loss of CENP-F would affect neuronal differentiation. The work was assisted by an artificial intelligence-based algorithm that enabled us to perform an automated analysis of differentiation during time-lapse videorecording. We found that CENP-F undergoes a striking relocalisation from the nucleus to the cytoplasm and neuritic extensions during differentiation. CENP