Investigating the role of protein CENP-F in cellular models of neuronal differentiation over time.

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-F silencing yields a disorganised differentiation process. These data suggest therefore a requirement for CENP-F in neurodifferentiation.

On the 10th-11th of October a microscopy course for the PhD students of the BeMM School will be held at the IBPM-CNR imaging platform (BBCD Department, Polo San Lorenzo).

As detailed in the attached programme, the course will include a theoretical session open to all students; a practical session of confocal spinning disk 3D cell imaging (acquisition and analysis) open to 12 students in presence (divided in subgroups, 2 hours each) and to all interested students from remote. Samples will be provided by the imaging platform but there will be space for questions and discussion where students can explore the suitability of the instrumentation for applications of their interest.