Synthesis, storage and release of [14C]acetylcholine in isolated rat diaphragm muscles

作者：Lincoln T. Potter · 发表于：The Journal of Physiology · 年份：1970 · DOI：10.1113/jphysiol.1970.sp009003 · 被引用次数：360 · 研究领域：Neuroscience and Neuropharmacology Research、Nicotinic Acetylcholine Receptors Study、Neurological Disorders and Treatments

1. Segments of rat diaphragms were kept in choline-free media for 4 hr and were then exposed to a physiological concentration of [(14)C]-choline (30 muM) at 37 degrees C. The synthesis, storage and subsequent release of [(14)C]acetylcholine by the muscles was assessed by isotopic- and bio-assays after isolation of the transmitter by paper electrophoresis.2. Replacement of endogenous acetylcholine (0.92 mu-mole/kg) with labelled acetylcholine proceeded slowly at rest, but rapidly during nerve stimulation. [(14)C]Acetylcholine accumulated most rapidly when hydrolysis of the released transmitter, and thus the re-use of endogenous choline, was prevented by an esterase inhibitor. Fully replaced stores were maintained during nerve stimulation by synthesis rates sufficient to replenish at least 35% of the store size in 5 min.3 In the presence of hemicholinium-3, which inhibits choline uptake, acetylcholine stores declined rapidly during stimulation, and residual synthesis was slight, indicating little intraneural choline. Net choline uptake into nerve terminals was estimated from the highest observed synthesis rate and from previous measurements of the number and size of terminals, as 3-6 p-mole/cm(2) sec.4. Transmitter synthesis was localized in the region of end-plates, and was reduced to a few per cent of normal 6 weeks after phrenic nerve section. Release experiments suggested that at least half of the acetylcholine in phrenic nerves is in their terminals; from this content and ...