Fast axonal transport of the vesicular acetylcholine transporter (VAChT) in cholinergic neurons in the rat sciatic nerve

Jia Yi Li, Anna Maria Dahlström, Louis B. Hersh, Annica Dahlström

Research output: Contribution to journalArticlepeer-review

13 Scopus citations

Abstract

The sciatic nerve, as a part of the peripheral nervous system (PNS), has been used to study axonal transport for decades. It contains motor, sensory as well as autonomic axons. The present study has concentrated on the axonal transport of the synaptic vesicle acetylcholine transporter (VAChT), using the 'stop-flow/nerve crush' method. After blocking fast axonal transport by means of a crush, distinct accumulations of various synaptic vesicle proteins, including VAChT, and peptides developed during the first hour after crush-operation and marked increases were observed up to 8 h post-operative. Semiquantitative analysis, using cytofluorimetric scanning (CFS) of immuno-incubated sections, revealed a rapid rate of accumulation proximal to the crush, and that the ratio between distal accumulations (organelles in retrograde transport) and proximal accumulations (organelles in anterograde transport) was about 40%. Most synaptic vesicle proteins were colocalized in the axons proximal to the crush. VAChT-immunoreactive axons were also immunoreactive for choline acetyltransferase (ChAT). Autonomic axons with VAChT also contained VIP-LI. The results demonstrate (1) that VAChT, as well as other synaptic vesicle proteins, is transported with fast axonal transport in motor axons as well as in autonomic post-ganglionic neurons in this nerve, (2) VAChT colocalized in motor axons with SV2 as well as with synaptophysin, indicating storage in the same axonal particle, (3) in the autonomic postganglionic sympathetic cholinergic fibres, VAChT colocalized with VIP, but VIP-LI was present in rather large granular structures while VAChT-LI was present mostly as small granular elements, (4) in motor as well as in autonomic axons ChAT-LI was present in VAChT-positive axons, and (5) the ratio of recycling (retrogradely accumulated) VAChT-IR was about 40%, in contrast to the recycling fraction of synaptophysin that was about 70%.

Original languageEnglish
Pages (from-to)457-467
Number of pages11
JournalNeurochemistry International
Volume32
Issue number5-6
DOIs
StatePublished - May 1 1998

Bibliographical note

Funding Information:
Supported by the Swedish MRC (2207) , EU grant no 96 1586; the Royal Academy of Science and Arts in Göteborg, Gustav V:s 80-årsfond, and by the Swedish Society for Medical Research. For a generous supply of antibody preparations we are grateful to Dr R. Jahn, New Haven (p38) , Dr R. Kelly, San Francisco (SV2) , and Dr M. Goldstein, New York (anti-TH) . The technical assistance of Ms Kerstin Lundmark is gratefully acknowledged. Dahlstrom et al., 1992 Dahlstrom et al. 1978

Keywords

  • Axonal transport
  • Choline acetyltransferase
  • Confocal microscopy
  • Cytofluorimetric scanning
  • Immunofluorescence
  • Rat
  • Sciatic nerve
  • Synaptic vesicle proteins
  • VIP
  • Vesicular acetylcholine transporter (vacht)

ASJC Scopus subject areas

  • Cellular and Molecular Neuroscience
  • Cell Biology

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