title: Regulation of citrate metabolism and acetycholine synthesis by Ca(2+) in rat brain synaptosomes.
abstract: The relationships between pyruvate and derived citrate metabolism and acetylcholine (ACh) synthesis in synaptosomes were examined. In the presence of 30 mM KCl, 0.1 mM Ca(2+) caused 31 and 63% inhibition of pyruvate utilization and citrate accumulation, respectively. Verapamil and EGTA (0.5 mM) brought about no change in pyruvate consumption but increased rate of citrate accumulation, and overcame inhibitory effect of Ca(2+). The rates of citrate accumulation in the presence of verapamil or EGTA were three to six times, respectively, higher than those in the presence of Ca(2+). (?) Hydroxycitrate increased rate of citrate accumulation under all experimental conditions. The value of this activation appeared to be stable (0.20-0.28 nmol/min/mg of protein) and independent of changes in the basic rate of citrate accumulation. Ca(2+) caused no significant changes in [(14)C]ACh synthesis, but it inhibited (14)CO(2) production by synaptosomes. These activities were inhibited by verapamil by 33 and 60%, respectively. Ca(2+) did not modify these effects of the drug. On the other hand, (?)hydroxycitrate resulted in 22 and 29% inhibition of [(14)C]ACh synthesis in Ca(2+) free and Ca(2+) supplemented medium, respectively. These data indicated that rates of acetyl-CoA synthesis in synaptoplasm, via ATP-citrate lyase and probably by another pathways are independent of Ca-evoked changes in pyruvate oxidation and citrate supply from intraterminal mitochondria. This property might play a significant role in maintenance of stable level of ACh in active cholinergic nerve endings.
authors.affiliation: Department of Clinical Biochemistry, Medical Academy, De?binki 7, 80-211 Gda?sk, Poland.
authors.forename: A
authors.initials: A
authors.lastname: Szutowicz
authors.forename: H
authors.initials: H
authors.lastname: Bielarczyk
authors.forename: D
authors.initials: D
authors.lastname: Sosnowska
authors.forename: B
authors.initials: B
authors.lastname: Ciszek