Developmental timing-dependent organization of synaptic connections between mossy fibers and granule cells in the cerebellum

The long-standing hypothesis that synapses between mossy fibers (MFs) and cerebellar granule cells (GCs) are organized according to the origins of MFs and locations of GC axons, parallel fibers (PFs), is supported by recent findings. However, the mechanisms of such organized synaptic connections remain unknown. Here, using our technique that enabled PF location-dependent labeling of GCs in mice, we confirmed that synaptic connections of GCs with specific MFs originating from the pontine nucleus (PN-MFs) and dorsal column nuclei (DCoN-MFs) were gently but differentially organized according to their PF locations. We then found that overall MF-GC synaptic connectivity was biased in a way that dendrites of GCs having nearby PFs tended to connect with the same MF terminals, implying that the MF origin- and PF location-dependent organization is associated with the overall biased MF-GC synaptic connectivity. Furthermore, the development of PN-MFs preceded that of DCoN-MFs, which matches the developmental sequence of GCs that preferentially connect with each type of these MFs. Thus, our results revealed that overall MF-GC synaptic connectivity is biased in terms of PF locations, and suggested that such connectivity is likely the result of synaptic formation between developmental timing-matched partners. Parallel fiber location-dependent labeling of granule cells in the mouse cerebellum shows that their synaptic connections with presynaptic mossy fibers are likely to be the result of matched developmental timing.