Recovery of [3H]acetylcholine synthesis after AF64A-treatment in primary, neuron-enriched, rat septal-hippocampal and striatal cultures

Brain Res Dev Brain Res. 1993 Jul 16;74(1):51-6. doi: 10.1016/0165-3806(93)90082-l.


Neuron-enriched cultures prepared from several different rat brain regions were incubated with 10 microM or 30 microM monoethylcholine mustard aziridinium ion (AF64A) under conditions (1 h, 37 degrees C in Krebs Ringer buffer) that reduced acetylcholine (ACh) synthesis coupled to high-affinity choline uptake, without affecting choline acetyltransferase activity. Co-cultures of septum-hippocampus and cultures of striatum were similarly sensitive to the AF64A-induced inhibition of ACh synthesis. However, ACh-synthesis recovered more rapidly in the striatal cultures than in septal-hippocampal co-cultures after AF64A washout. In septal-hippocampal co-cultures, neither tunicamycin (20 micrograms/ml) nor cycloheximide (0.5 microgram/ml) had any effect on the basal synthesis of ACh or its recovery within 24 h following 10 microM AF64A washout. However, the recovery of ACh synthesis in these co-cultures after 30 microM AF64A-washout was blocked by either tunicamycin or cyclohexamide. Neither tunicamycin nor cyclohexamide interfered with ACh-synthesis recovery after washout of 30 microM AF64A in striatal cultures. These studies suggest that the turnover of high-affinity choline transporters can be modulated in a brain-region specific manner in intact primary neuronal cultures.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Acetylcholine / biosynthesis*
  • Animals
  • Aziridines / pharmacology*
  • Cell Count / drug effects
  • Cells, Cultured
  • Choline / analogs & derivatives*
  • Choline / metabolism
  • Choline / pharmacology
  • Choline O-Acetyltransferase / drug effects
  • Choline O-Acetyltransferase / metabolism
  • Corpus Striatum / cytology
  • Corpus Striatum / drug effects*
  • Glycosylation
  • Hippocampus / cytology
  • Hippocampus / drug effects*
  • Nerve Tissue Proteins / biosynthesis
  • Neurons / drug effects*
  • Rats
  • Rats, Sprague-Dawley
  • Septum Pellucidum / cytology
  • Septum Pellucidum / drug effects*
  • Tritium


  • Aziridines
  • Nerve Tissue Proteins
  • Tritium
  • ethylcholine aziridinium
  • Choline O-Acetyltransferase
  • Choline
  • Acetylcholine