Each of the projects described herein opened up fundamental questions that should be addressed. Unfortunately, practical considerations force us to set priorities. Our current efforts are centered on: 1) defining the molecular mechanism of myelin sheath formation by following the synthesis of MyMs, their packaging into MFOs and transportation along an oligodendrocyte process using live imaging; 2) purifying and characterizing the contents of MFOs. We are using 3D electron tomography of embryonic avian optic nerve at different stages of development to study the fusion of MyMs into a myelin sheath.

  The other project is to test the hypothesis that ectosomes are the conveyors of signals that coordinate and orchestrate the modular structure of the myelinated axon. We are developing technologies to obtain pure cell-specific ectosomes by molecularly color-code them. We are also customizing a microfluidic platform suitable to integrate multiple processes in a single lab-on-a-chip format. We plan to establish unique EM grid compatible bi-chamber for OLG-neuron cultures and capture the formation of cell-specific ectosomes using correlative light and electron microscopy and phase contrast cryo-electron tomography. We will assemble tomograms of the initial OLG-neuron encounter so that we can structurally model the interface in a 3D space.