The Journal of Membrane Biology

, Volume 29, Issue 1, pp 47–65 | Cite as

Potentiation of postjunctional cholinergic sensitivity of rat diaphragm muscle by high-energy-phosphate adenine nucleotides

  • Douglas A. Ewald


The cholinergic sensitivity of rat diaphragm muscle, measured as the magnitude of depolarization responses to repetitive, iontophoretic pulses of acetylcholine (ACh) onto neuromuscular endplates, is increased by addition of ATP to the perfusion medium. Depolarization responses begin to increase within the first min after addition of 10mm ATP and plateau at 60% above control levels (mean value) after 4 to 6 min. Neither the magnitude nor the time course of the potentiations corresponds to changes in resting potential or membrane resistance. Other nucleotides are equally or less effective at the same concentration: ATP-ADP>UTP>AMP=GTP (=no added nucleotide control) The duration of the individual ACh responses does not increase during continuous exposure to the active nucleotides for up to 15 min except when the muscle is pretreated with eserine.

Mild enzymatic predigestion of the muscle with collagenase and then protease, increasing the availability of the postjunctional membrane to bath-applied drugs, decreases the variability and increases the magnitude of the potentiation to a given dose of ATP. The dose-response curve for ATP is then more than half-maximal at 1mm and the ranking of the other nucleotides relative to ATP is the same as without predigestion.

There is an optimum Ca++ concentration for the potentiation between zero and 2mm: potentiation is enhanced in Ca++-free medium, partially blocked in twice-normal Ca++ medium, and totally blocked in Ca++-free medium 10 min after a 5 min exposure to 2.5mm EGTA. The similar Ca++ dependence of ACh receptor activation in the absence of added nucleotide suggests that ATP directly facilitates receptor activation by ACh. This facilitory action could be one of the physiological roles for the ATP released from stimulated phrenic nerve.


Free Medium Phrenic Nerve Adenine Nucleotide Membrane Resistance Perfusion Medium 
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Copyright information

© Springer-Verlag New York Inc. 1976

Authors and Affiliations

  • Douglas A. Ewald
    • 1
  1. 1.Department of ZoologyUniversity of CaliforniaBerkeley

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