Elise LV Malavasi,1 Aniket Ghosh,1 Daniel G Booth,2 Michele Zagnoni,3 Diane L Sherman,1 and Peter J Brophy1


1Centre for Discovery Brain Sciences, University of Edinburgh, Edinburgh, United Kingdom
2Biodiscovery Institute, University of Nottingham, Nottingham, United Kingdom
3Centre for Microsystems & Photonics, Dept. Electronic and Electrical Engineering, University of Strathclyde, Strathclyde, United Kingdom
Peter J Brophy: [email protected]




Voltage-gated sodium channels cluster in macromolecular complexes at nodes of Ranvier to promote rapid nerve impulse conduction in vertebrate nerves. Node assembly in peripheral nerves is thought to be initiated at heminodes at the extremities of myelinating Schwann cells, and fusion of heminodes results in the establishment of nodes. Here we show that assembly of ‘early clusters’ of nodal proteins in the murine axonal membrane precedes heminode formation. The neurofascin (Nfasc) proteins are essential for node assembly, and the formation of early clusters also requires neuronal Nfasc. Early clusters are mobile and their proteins are dynamically recruited by lateral diffusion. They can undergo fusion not only with each other but also with heminodes, thus contributing to the development of nodes in peripheral axons. The formation of early clusters constitutes the earliest stage in peripheral node assembly and expands the repertoire of strategies that have evolved to establish these essential structures.



The microscope was equipped with a x10/0.3 NA Plan Fluor long working distance objective lens and a Plan Achromat x100/1.10 NA water-immersion objective lens, Chroma ET filters, an Orca Flash 4.0v3 sCMOS camera (Hammamatsu Photonics), and LED illumination (CoolLED pE-4000) controlled by NIS-Elements AR software.

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The pE-4000 Universal Illumination System offers 16 selectable wavelengths from 365 - 770 nm, making it a highly flexible illuminator covering a wide variety of fluorophores

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