Inhibition by hemicholinium-3 of (14C)acetylcholine synthesis and (3H)choline high-affinity uptake in rat striatal synaptosomes.

Hemicholinium (HC-3), a neuromuscular blocking agent which inhibits the synthesis and the release of acetylcholine (ACh) was investigated for its effects both on [14C]ACh synthesis from [2-14C]pyruvate or [6-14C]glucose and on [3H]choline uptake in purified rat striatal synaptosomes. The synthesis of the transmitter was reduced to 15% of control by 1 µM HC-3 in the presence of eserine (170 µM). The drug produced half-maximal inhibition at 0.06 µM; this effect was totally reversed by 30 µM choline. Choline concentrations as low as 10 µM stimulated significantly the synthesis of [14C]ACh in a diluted suspension of synaptosomes. The existence of a high-affinity uptake system for choline, recently observed by Yamamura and Snyder, was confirmed in our preparation: its Km was found to be 3.5 µM. Eserine (170 µM) produced only 10% inhibition of the uptake process in the presence of 1 µM choline. HC-3 was found to be a purely competitive inhibitor of the choline high-affinity uptake ( Ki = 25 nM). A very good correlation was found between the inhibitory effects of HC-3 on [14C]ACh synthesis and on [3H]choline high-affinity uptake. The results suggest that the high-affinity system for the transport of choline is present in cholinergic synaptosomes and plays an important role in sustaining and perhaps regulating the synthesis of the transmitter. ACKNOWLEDGMENT The authors are very grateful to Dr. M. Israel for constant help, advice, and stimulating discussions.

• Publication year

  1973

• Venue

  Molecular Pharmacology

• Publication date

  1973-09-01

• Fields of study

  Medicine, Chemistry

• Identifiers
  DOI 10.1016/s0026-895x(25)13987-4PMID 4788157
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• Inhibition of [14C]acetylcholine synthesis closely correlated with inhibition of choline high-affinity uptake, supporting a role for the transport system in sustaining transmitter synthesis in cholinergic synaptosomes.

  Confidence 0.93

  mexicorea (qjvnbu8xg3) extractionAnonymous (12632b8b5f) review
• The choline high-affinity uptake system in rat striatal synaptosomes had Km = 3.5 µM, and eserine caused only about 10% inhibition of uptake at 1 µM choline.

  Confidence 0.95

  mexicorea (qjvnbu8xg3) extractionAnonymous (12632b8b5f) review
• Hemicholinium-3 was a purely competitive inhibitor of choline high-affinity uptake, with Ki = 25 nM.

  Confidence 0.97

  mexicorea (qjvnbu8xg3) extractionAnonymous (12632b8b5f) review
• 1 µM hemicholinium-3 reduced [14C]acetylcholine synthesis to 15% of control in the presence of eserine, with half-maximal inhibition at 0.06 µM and reversal by 30 µM choline.

  Confidence 0.98

  mexicorea (qjvnbu8xg3) extractionAnonymous (12632b8b5f) review

• acetylcholine synthesis

  process

  The formation of acetylcholine from labeled metabolic precursors measured in purified rat striatal synaptosomes.

  Aliases: ACh synthesis, [14C]ACh synthesis

  mexicorea (qjvnbu8xg3) extractionAnonymous (12632b8b5f) review
• choline

  compound

  The endogenous precursor and transport substrate supplied at micromolar concentrations in the assays.

  mexicorea (qjvnbu8xg3) extractionAnonymous (12632b8b5f) review
• choline high-affinity uptake

  transport process

  The saturable, high-affinity transport of choline into synaptosomes measured with [3H]choline.

  Aliases: high-affinity choline uptake, [3H]choline uptake

  mexicorea (qjvnbu8xg3) extractionAnonymous (12632b8b5f) review
• eserine

  reagent

  An acetylcholinesterase inhibitor added during assays to limit acetylcholine breakdown.

  mexicorea (qjvnbu8xg3) extractionAnonymous (12632b8b5f) review
• hemicholinium-3

  compound

  A choline-transport blocking drug used in the synaptosome assays to probe acetylcholine synthesis and uptake.

  Aliases: HC-3, hemicholinium

  mexicorea (qjvnbu8xg3) extractionAnonymous (12632b8b5f) review
• rat striatal synaptosomes

  biological sample

  Purified nerve-terminal preparations from rat striatum used as the experimental system.

  Aliases: purified rat striatal synaptosomes, striatal synaptosomes

  mexicorea (qjvnbu8xg3) extractionAnonymous (12632b8b5f) review

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