Assessment of choline uptake for the synthesis and release of acetylcholine in brain slices by a dynamic autoradiographic technique using [11C]choline.
The uptake of choline for the synthesis and release of acetylcholine was investigated in brain slices by dynamic positron autoradiography using [11C]choline. Brain slices (330 microm) were incubated with [11C]choline in oxygenated Krebs-Ringer medium at 34 degrees C and serial two-dimensional time-resolved images of the uptake and release of radioactivity were recorded on Storage Phosphor screens. [11C]choline uptake increased with the period of incubation and was 1.9 times higher in the striatum than cerebral cortex. The uptake in the striatum was significantly diminished by hemicholinium-3 (HC-3), an inhibitor of high-affinity choline uptake. Pretreatment of brain slices with 50 mM K(+) for 20 min enhanced the uptake in striatum. The uptake of [11C]choline in brain slices was saturable using nonlabeled choline. Two uptake systems, a high-affinity and a low-affinity system, were confirmed to exist by kinetic analysis using Lineweaver-Burk plots. The 11C radioactivity that had accumulated in the striatum disappeared on treatment with veratridine, a depolarization agent, in the presence of HC-3. This pattern of disappearance was consistent with that of the appearance of unlabeled and labeled acetylcholine in the medium. These results indicate that this method is useful for obtaining information regarding the uptake of choline for the synthesis and release of acetylcholine in live brain tissues.
Sasaki
Toru
T
Positron Medical Center, Tokyo Metropolitan Institute of Gerontology, 35-2 Sakae-cho, Itabashi-ku, Tokyo 173-0015, Japan. sasaki@pet.tmig.or.jp
Kawamura
Kazunori
K
Tanaka
Yasukazu
Y
Ando
Susumu
S
Senda
Michio
M
D000109:Acetylcholine / Q000378:metabolism*; D000818:Animals; D001345:Autoradiography / Q000295:instrumentation / Q000379:methods*; D001921:Brain / Q000187:drug effects / Q000378:metabolism*; D002250:Carbon Radioisotopes / Q000378:metabolism; D002540:Cerebral Cortex / Q000187:drug effects / Q000378:metabolism; D002794:Choline / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D003342:Corpus Striatum / Q000187:drug effects / Q000378:metabolism; D004583:Electrons; D004586:Electrophoresis; D006426:Hemicholinium 3 / Q000494:pharmacology; D007700:Kinetics; D008297:Male; D014179:Neurotransmitter Uptake Inhibitors / Q000494:pharmacology; D011188:Potassium / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D014701:Veratridine / Q000494:pharmacology
D002250:Carbon Radioisotopes; D014179:Neurotransmitter Uptake Inhibitors; D006426:Hemicholinium 3; D014701:Veratridine; D002794:Choline; D000109:Acetylcholine; D011188:Potassium