Release of Full-Length EphB2 Receptors from Hippocampal Neurons to Cocultured Glial Cells

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Abstract
Glial cells are known to actively participate in neuronal development by shaping neuronal connections through axon pruning and by controlling dendritic spine morphology. These functions may in part be mediated by engulfment of neuronal structures and trans-endocytosis of neuronal material into glial cells. These processes are not well understood, and the molecular components that mediate these events have primarily been elusive. Here, we implicate the Eph/ephrin signaling system in trans-endocytosis events at the neuron-to-glia interface. Using time-lapse microscopy, we show that hippocampal neurons exogenously expressing EphB2 receptors release or pinch-off EphB2-containing vesicles at sites of neuron-to-glia contact. Cocultured glial cells endogenously express the corresponding ephrinB ligands and are able to trans-endocytose full-length EphB2 from neighboring cells. Although Eph/ephrin signaling often occurs in a bidirectional manner, the observed vesicle release from neurons to glia was only observed in a unidirectional manner, i.e., when the neurons expressed EphB2, but not ephrinBs. These findings suggest that Eph/ephrin signaling is involved in the glial cell-mediated fine sculpting of neuronal structures.