Abstract
Patch-clamp recordings of current through acetylcholine-activated channels were made from non-junctional membrane of innervated myotomal muscle from Xenopus laevis. Two classes of acetylcholine (ACh) receptor channels were identified on the basis of current amplitudes. Both amplitude classes exhibited current-voltage relations which deviated from linearity as the extrapolated reversal potential was approached (-5 to -12 mV). Over the range of greatest linearity the conductances of the two classes were 64 and 44 pS. Both event classes were blocked by alpha-bungarotoxin. At the normal resting membrane potential (approximately -95 mV) the larger conductance channel (gamma) exhibited an apparent mean channel open time of less than 1 ms, compared to approximately 2 ms for the smaller gamma class. The apparent open time was voltage-dependent, changing e-fold with a 63 mV hyperpolarization for the high gamma channel and 93 mV hyperpolarization for the low gamma channel. At low ACh concentrations (0.1-0.3 microM) both amplitude classes exhibited bursts of successive openings separated by brief closures of less than 0.5 ms. Bursts were separated by longer closed intervals of 1 to greater than 100 ms. Closed interval histograms revealed corresponding populations of brief and long closures, indicating that at least two kinetic processes are required to describe the distribution of closed intervals. In the absence of exogenous ACh, channels were observed in an occasional patch which showed a conductance and extrapolated reversal potential similar to ACh-activated channels. In such patches the event frequency could occasionally be altered by adjusting the negative pressure applied to the patch. The two main conductance classes of ACh activated channels were observed to coexist in most patches. However, the most frequent event observed in non-junctional membrane of innervated muscle corresponded to the high gamma class. In this respect, the non-junctional ACh receptors bore a greater similarity to junctional ACh receptors than to non-junctional receptors reported for denervated muscle.
MeSH Terms
Acetylcholine/pharmacology
Action Potentials/drug effects
Animals
Bungarotoxins/pharmacology
In Vitro Techniques
Ion Channels/physiology
Kinetics
Membrane Potentials/drug effects
Muscles/innervation
Receptors, Cholinergic/physiology
Time Factors
Xenopus laevis
Chemicals
Bungarotoxins
Ion Channels
Receptors, Cholinergic
Acetylcholine
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Brehm P
Kullberg R
Moody-Corbett F
References (25)
25 references, click to expand
-
The statistical nature of the acetycholine potential and its molecular components.
J Physiol. 1972 Aug;224(3):665-99
PMID: 5071933
-
Developmental changes in the distribution of acetylcholine receptors in the myotomes of Xenopus laevis.
J Physiol. 1983 Jun;339:553-71
PMID: 6887034
-
Fluorescent staining of acetylcholine receptors in vertebrate skeletal muscle.
J Physiol. 1974 Mar;237(2):385-400
PMID: 4133039
-
Single-channel currents recorded from membrane of denervated frog muscle fibres.
Nature. 1976 Apr 29;260(5554):799-802
PMID: 1083489
-
Noise analysis of drug induced voltage clamp currents in denervated frog muscle fibres.
J Physiol. 1976 Jul;258(3):705-29
PMID: 1086359
-
Junctional and extrajunctional acetylcholine receptors in normal and denervated frog muscle fibres. Noise analysis experiments with different agonists.
Pflugers Arch. 1976 Oct 15;366(1):1-9
PMID: 185583
-
Development of the myotomal neuromuscular junction in Xenopus laevis: an electrophysiological and fine-structural study.
Dev Biol. 1977 Oct 1;60(1):101-29
PMID: 902924
-
Acetylcholine-induced ionic channels in rat skeletal muscle.
Fed Proc. 1978 Oct;37(12):2654-9
PMID: 212327
-
Change in synaptic channel gating during neuromuscular development.
Nature. 1978 Nov 23;276(5686):401-2
PMID: 714165
-
Single acetylcholine-activated channels show burst-kinetics in presence of desensitizing concentrations of agonist.
Nature. 1980 Jul 3;286(5768):71-3
PMID: 6248795
-
A post-natal decrease in acetylcholine channel open time at rat end-plates.
J Physiol. 1980 Jun;303:125-37
PMID: 6253617
-
Receptor stability and channel conversion in the subsynaptic membrane of the developing mammalian neuromuscular junction.
Dev Biol. 1980 Nov;80(1):1-17
PMID: 6254823
-
Nonjunctional acetylcholine receptor channel open time decreases during development of Xenopus muscle.
Nature. 1981 Jan 29;289(5796):411-3
PMID: 6258078
-
Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches.
Pflugers Arch. 1981 Aug;391(2):85-100
PMID: 6270629
-
Multiple conductance states of single acetylcholine receptor channels in embryonic muscle cells.
Nature. 1981 Dec 3;294(5840):462-4
PMID: 6273742
-
Fluctuations in the microsecond time range of the current through single acetylcholine receptor ion channels.
Nature. 1981 Dec 3;294(5840):464-6
PMID: 6273743
-
Inward current channels activated by intracellular Ca in cultured cardiac cells.
Nature. 1981 Dec 24;294(5843):752-4
PMID: 6275271
-
Single Ca2+-activated nonselective cation channels in neuroblastoma.
Nature. 1982 Mar 25;296(5855):357-9
PMID: 6278324
-
Channel open time of acetylcholine receptors on Xenopus muscle cells in dissociated cell culture.
Dev Biol. 1982 May;91(1):93-102
PMID: 6284571
-
Single-channel currents in isolated patches of plasma membrane from basal surface of pancreatic acini.
Nature. 1982 Sep 9;299(5879):159-61
PMID: 6287285
-
Acetylcholine receptor kinetics. A description from single-channel currents at snake neuromuscular junctions.
Biophys J. 1982 Sep;39(3):253-61
PMID: 6291654
-
Metabolism of acetylcholine receptors on embryonic amphibian muscle.
J Neurosci. 1983 Jan;3(1):101-7
PMID: 6822849
-
Successive openings of the same acetylcholine receptor channel are correlated in open time.
Biophys J. 1983 Apr;42(1):109-14
PMID: 6301575
-
Neurotrophic control of channel properties at neuromuscular synapses of rat muscle.
J Physiol. 1983 Apr;337:159-71
PMID: 6875926
-
Voltage clamp analysis of acetylcholine produced end-plate current fluctuations at frog neuromuscular junction.
J Physiol. 1973 Dec;235(3):655-91
PMID: 4543940