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J Neurophysiol (April 1, 2003). 10.1152/jn.00668.2002
Submitted on Submitted 13 August 2002; accepted in final form 27 November 2002
1The Otto Loewi Minerva Center for Cellular and Molecular Neurobiology, Department of Neurobiology, The Hebrew University, Jerusalem 91904, Israel; 2Laboratory of Bioorganic Chemistry, National Institute of Diabetes and Digestive and Kidney Diseases, Bethesda, Maryland 20892; and 3Lehrstuhl für Zelluläre Physiologie, Ludwig-Maximilians-Universität, 80336 Munich, Germany
Slutsky, I.,
J. Wess,
J. Gomeza,
J. Dudel,
I. Parnas, and
H. Parnas.
Use of Knockout Mice Reveals Involvement of
M2-Muscarinic Receptors in Control of the Kinetics of
Acetylcholine Release. J. Neurophysiol. 89: 1954-1967, 2003. We have previously suggested
that presynaptic M2-muscarinic receptors
(M2R) are involved in the control of the time
course of evoked acetylcholine release in the frog neuromuscular
junction. The availability of knockout mice lacking functional
M2R (M2-KO) enabled us to
address this issue in a more direct way. Using the phrenic diaphragm
preparation, we show that in wild-type (WT) mice experimental
manipulations known to affect Ca2+ entry and
removal, greatly affected the amount of acetylcholine released (quantal
content). However, the time course of release remained unaltered under
all these experimental treatments. On the other hand, in the
M2-KO mice, similar experimental treatments affected both the quantal content and the time course of release. In
general, a larger quantal content was accompanied by a longer duration
of release. Similarly, the rise time of the postsynaptic current
produced by axon stimulation was sensitive to changes in
[Ca2+]o or
[Mg2+]o in
M2-KO mice but not in WT mice. Measurements of
Ca2+ currents revealed that the shorter rise time
of the postsynaptic current seen in high
[Mg2+]o in
M2-KO mice was not produced by a shorter wave of
the presynaptic Ca2+ current. These results
support our earlier findings and provide direct evidence for the major
role that presynaptic M2-muscarinic receptors
play in the control of the time course of evoked acetylcholine release
under physiological conditions.
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