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Alternative splicing is crucial for molecular diversification, which greatly contributes to the complexity and specificity of neural functions in the central nervous system (CNS). Neurofascin (NF) is a polymorphic cell surface protein that has a number of splicing isoforms. As the alternative splicing of the neurofascin gene ( Nfasc ) is developmentally regulated, NF isoforms have distinct functions in immature and mature brains. However, the molecular mechanisms underlying the alternative splicing of  Nfasc  in neurons are not yet understood. Here, we demonstrate that, alongside developmental regulation,  Nfasc  alternative splicing is spatially controlled in the mouse brain. We then identified distinct  Nfasc  splicing patterns at the cell-type level in the cerebellum, with  Nfasc186  being expressed in Purkinje cells and absent from granule cells (GCs). Furthermore, we show that high K + -induced depolarization triggers a shift in splicing from  Nfasc140  to  Nfasc186  in cerebellar GCs. Finally, we identified a neural RNA-binding protein, Rbfox, as a key player in neural NF isoform selection, specifically controlling splicing at exons 26−29. Together, our results show that  Nfasc  alternative splicing is spatio-temporally and dynamically regulated in cerebellar neurons. Our findings provide profound insight into the mechanisms underlying the functional diversity of neuronal cell-adhesive proteins in the mammalian CNS.

Spatio-temporal and dynamic regulation of neurofascin alternative splicing in mouse cerebellar neurons