Precursor and mature NGF live tracking: one versus many at a time in the axons

Scientific Reports
Teresa De NadaiAntonino Cattaneo

Abstract

The classical view of nerve growth factor (NGF) action in the nervous system is linked to its retrograde axonal transport. However, almost nothing is known on the trafficking properties of its unprocessed precursor proNGF, characterized by different and generally opposite biological functions with respect to its mature counterpart. Here we developed a strategy to fluorolabel both purified precursor and mature neurotrophins (NTs) with a controlled stoichiometry and insertion site. Using a single particle tracking approach, we characterized the axonal transport of proNGF versus mature NGF in living dorsal root ganglion neurons grown in compartmentalized microfluidic devices. We demonstrate that proNGF is retrogradely transported as NGF, but with a lower flux and a different distribution of numbers of neurotrophins per vesicle. Moreover, exploiting a dual-color labelling technique, we analysed the transport of both NT forms when simultaneously administered to the axon tips.

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Citations

Mar 12, 2017·International Journal of Molecular Sciences·Maria S Ioannou, Margaret Fahnestock
Sep 14, 2019·Proceedings of the National Academy of Sciences of the United States of America·Laura MarchettiAntonino Cattaneo
Mar 12, 2019·Frontiers in Neuroscience·Margaret Fahnestock, Arman Shekari
Nov 2, 2019·Traffic·Sunaina SuranaGiampietro Schiavo
Feb 18, 2017·Frontiers in Molecular Biosciences·Pierluigi Di MatteoAntonino Cattaneo
Nov 17, 2020·Frontiers in Neuroscience·Domenica ConvertinoCamilla Coletti

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Methods Mentioned

BETA
ion exchange chromatography
fluorescence microscopy

Software Mentioned

Matlab scripts

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