Two-dimensional gel electrophoresis of proteins in rapid axoplasmic transport

George C. Stone, David L. Wilson, Michael E. Hall

Research output: Contribution to journalArticle

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Abstract

Two-dimensional electrophoresis of proteins has allowed high resolution analysis of the protein species rapidly transported in the frog sciatic nerve. The 7th, 8th and 9th dorsal root ganglia were selectively labeled with [3H]leucine or [35S]methionine in one compartment of a lucite chamber. Transport of TCA-precipitable material was monitored in the spinal roots and sciatic nerve kept in another compartment. Fastest transport rates were 75-90 mm/day at 18 °C. Ligation of the nerve 30 mm distal to the 8th ganglion at the beginning of the experiment resulted in accumulation of label during a 24 h period. This material was subjected to two-dimensional electrophoresis (pI 5-8; mol.wt. 104-105 daltons) in 3 mm nerve segments. Autoradiographs or fluorographs from segments proximal to the ligature yielded a pattern of about 140 spots. Of these, at least 60 were considered to be independent protein species. Neither actin nor tubulin were present among these rapidly-transported, labeled proteins. No pattern was observed from segments distal to the ligature. Blocking protein synthesis with 18 μM anisomycin reduced the accumulation of label proximal to the ligature by 98%. Direct labeling of nerve segments produced patterns significantly different from the pattern of transported proteins.

Original languageEnglish (US)
Pages (from-to)287-302
Number of pages16
JournalBrain Research
Volume144
Issue number2
DOIs
StatePublished - Apr 14 1978

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ASJC Scopus subject areas

  • Neuroscience(all)
  • Molecular Biology
  • Clinical Neurology
  • Developmental Biology

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