Pharmacological characteristics of choline transport system in mouse cerebral cortical neurons in primary culture.
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概要
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The characteristics of [<SUP>3</SUP>H]choline transport with high affinity were investigated using primary cultured neurons obtained from the mouse cerebral cortex. [<SUP>3</SUP>H]Choline uptake was saturable as a function of extracellular [<SUP>3</SUP>H]choline concentration. Analysis by Lineweaver-Burk plot revealed that [<SUP>3</SUP>H]choline was transported into neurons by a high affinity transport system with a K<SUB>m</SUB> value of 19.8 ± 0.8μM and V<SUB>max</SUB> value of 0.334 plusmn; 0.022 nmol/mg protein/min. This high affinity transport of [<SUP>3</SUP>H]choline was significantly inhibited by the withdrawal of sodium from the incubation medium, incubation at low temperature (4°C) and addition of metabolic inhibitors such as monoiodoacetate. These results indicate that the high affinity [<SUP>3</SUP>H]choline uptake in primary cultured neurons is sodium- and energy-dependent. Hemicholinium-3 also showed a competitive inhibition on the [<SUP>3</SUP>H]choline transport. Depolarization by high K<SUP>+</SUP> induced an enhancement of the [<SUP>3</SUP>H]choline uptake in the presence of Ca<SUP>2+</SUP>. The crude synaptosomal fraction obtained from primary cultured neurons possessed approximately forty-fold higher synthesizing activity of [<SUP>3</SUP>H]acetylcholine from [<SUP>3</SUP>H]choline than that found in the homogenate preparation of cultured neurons. The present results strongly suggest that the primary cultured neurons used in this study possess a sodium- and energy-dependent high-affinity choline uptake system as well as a synthesizing system for acetylcholine. Possible usefulness of these neurons for investigating neuronal uptake of choline and its functional role in the biosynthesis of acetylcholine are also suggested.
- 公益社団法人 日本薬理学会の論文
著者
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Ohkuma Seitaro
Department Of Pharmacology Kawasaki Medical School
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Kishi Masataka
Department Of Partial Prosthodontics Tokyo Dental College
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Kuriyama Kinya
Department Of Health Promoting Acupuncture And Moxibution Meiji University Of Oriental Medicine
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Ma Fu-Hai
Department of Pharmacology, Kyoto Prefectural University of Medicine
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Kishi Masataka
Department of Pharmacology, Kyoto Prefectural University of Medicine
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