Recovery of [3H]acetylcholine synthesis after AF64A-treatment in primary, neuron-enriched, rat septal-hippocampal and striatal cultures.
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.
Meyer
E M
EM
Department of Pharmacology and Therapeutics, University of Florida College of Medicine, Gainesville 32610-0267.
Judkins
J H
JH
Hardwick
E O
EO
D000109:Acetylcholine / Q000096:biosynthesis*; D000818:Animals; D001388:Aziridines / Q000494:pharmacology*; D002452:Cell Count / Q000187:drug effects; D002478:Cells, Cultured; D002794:Choline / Q000031:analogs & derivatives* / Q000378:metabolism / Q000494:pharmacology; D002795:Choline O-Acetyltransferase / Q000187:drug effects / Q000378:metabolism; D003342:Corpus Striatum / Q000166:cytology / Q000187:drug effects*; D006031:Glycosylation; D006624:Hippocampus / Q000166:cytology / Q000187:drug effects*; D009419:Nerve Tissue Proteins / Q000096:biosynthesis; D009474:Neurons / Q000187:drug effects*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012688:Septum Pellucidum / Q000166:cytology / Q000187:drug effects*; D014316:Tritium
D001388:Aziridines; D009419:Nerve Tissue Proteins; D014316:Tritium; C044894:ethylcholine aziridinium; D002795:Choline O-Acetyltransferase; D002794:Choline; D000109:Acetylcholine