The packing of acetylcholine into quanta at the frog neuromuscular junction is inhibited by increases in intracellular sodium.
Pretreatment with hypertonic solutions, insulin, or adrenaline increases the size of quanta at the frog neuromuscular junction, as determined by measurements of miniature end plate potentials or currents (Van der Kloot and Van der Kloot 1985, 1986). The increase in quantal size apparently is due to an increase in acetylcholine (ACh) content of individual quanta. These treatments, therefore, can be used to study the packaging of ACh. Previously, I reported that increases are blocked by an inhibitor of active ACh uptake into vesicles (Van der Kloot 1986b, 1987b). The present study shows that the increases in quantal size were antagonized by inhibiting the Na+-K+ exchange pump with 100 microM ouabain, 10 microM dihydroouabain, or K+-free solutions. The increases in quantal size were also antagonized by 10 microM monensin, a Na+ ionophore, or by 5 microM aconitine, which opens Na+ channels at normal resting potentials. Apparently a rise in intracellular [Na+] inhibits the addition of ACh to quanta. The mechanism by which a rise in intracellular Na+ inhibits ACh packing is unknown, but apparently it is not due to inhibition of choline reuptake into the terminals. Also consistent with the above hypothesis is that the increase in quantal size following depolarization for 2 h in elevated [K+]out was substantially enhanced when tetrodotoxin (TTX) was present, suggesting that in the absence of TTX there is a rise in [Na+]in that antagonizes the incorporation of additional ACh into the quanta.
Van der Kloot
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Department of Physiology, H.S.C., SUNY, Stony Brook 11794.
D000109:Acetylcholine / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000818:Animals; D002794:Choline / Q000494:pharmacology; D005110:Extracellular Space; D005942:Gluconates / Q000494:pharmacology; D006982:Hypertonic Solutions; D007328:Insulin / Q000494:pharmacology; D007425:Intracellular Membranes / Q000378:metabolism*; D009469:Neuromuscular Junction / Q000378:metabolism*; D010042:Ouabain / Q000031:analogs & derivatives / Q000494:pharmacology; D011188:Potassium / Q000494:pharmacology; D011789:Quantum Theory; D011894:Rana pipiens; D012462:Saline Solution, Hypertonic / Q000494:pharmacology; D012964:Sodium / Q000172:deficiency / Q000378:metabolism*
D005942:Gluconates; D006982:Hypertonic Solutions; D007328:Insulin; D012462:Saline Solution, Hypertonic; C000977:dihydroouabain; D010042:Ouabain; D012964:Sodium; D002794:Choline; D000109:Acetylcholine; C030691:gluconic acid; D011188:Potassium