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

Teresa De Nadai, Laura Marchetti, Carmine Di Rienzo, Mariantonietta Calvello, Giovanni Signore, Pierluigi Di Matteo, Francesco Gobbo, Sabrina Turturro, Sandro Meucci, Alessandro Viegi, Fabio Beltram, Stefano Luin, Antonino Cattaneo

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish
Pages (from-to)20272
JournalScientific Reports
Volume6
DOIs
Publication statusPublished - 1 Feb 2016
Externally publishedYes

Keywords

  • Amino Acid Sequence
  • Animals
  • Axonal Transport
  • Axons/metabolism
  • Ganglia, Spinal
  • Humans
  • Models, Molecular
  • Nerve Growth Factor/chemistry
  • Neurons/metabolism
  • Protein Conformation
  • Protein Interaction Domains and Motifs
  • Protein Multimerization
  • Protein Precursors/chemistry
  • Rats
  • Transport Vesicles/metabolism

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