Postsynaptic nicotinic receptor desensitized by non-contractile Ca2+ mobilization via protein kinase-C activation at the mouse neuromuscular junction.
1. Non-contractile Ca2+ mobilization (unaccompanied by muscle contraction) was initiated by nerve stimulation in the presence of neostigmine (more than 0.03 microM) at the endplate region of mouse diaphragm muscles. In the process of nicotinic receptor desensitization, the depressant effect of non-contractile Ca2+ on contractile Ca2+ mobilization was investigated by measurement of Ca(2+)-aequorin luminescence. 2. When the phrenic nerve was stimulated with paired pulses having intervals of 150, 300, 600, 1000 and 2000 ms, contractile Ca2+ transients were elicited during the generation of non-contractile Ca2+ mobilization. The amplitude of the contractile Ca2+ transients elicited by the second pulse (S2) was depressed at the shorter pulse intervals, but recovered to the initial contractile response (S1) at longer pulse intervals. 3. The extent of depression of S2 was enhanced by increasing the concentration of neostigmine (0.03 to 0.3 microM). When a low concentration (0.05 microM) of pancuronium, a competitive nicotinic antagonist, completely blocked non-contractile Ca2+ mobilization, the depression of S2 was diminished. 4. The depression of S2 was enhanced when the peak amplitude of non-contractile Ca2+ mobilization was raised by increasing the external Ca2+ concentration from 1.3 to 5 mM. 5. Staurosporine (10 nM), a protein kinase-C inhibitor, diminished the depression of S2 despite large amounts of non-contractile Ca2+ mobilization. The diminishing effect of staurosporine was counteracted by TPA (0.1 microM), a protein kinase-C activator. 6. These findings suggest that non-contractile Ca2+ mobilization may enhance the desensitization of the postsynaptic nicotinic receptor via activation of protein kinase-C at the neuromuscular junction.
Kimura
I
I
Department of Chemical Pharmacology, Faculty of Pharmaceutical Sciences, Toyama Medical and Pharmaceutical University, Japan.
Dezaki
K
K
Tsuneki
H
H
Kimura
M
M
D000470:Alkaloids / Q000494:pharmacology; D000818:Animals; D002118:Calcium / Q000378:metabolism* / Q000494:pharmacology; D003964:Diaphragm / Q000294:innervation / Q000502:physiology; D004558:Electric Stimulation; D004789:Enzyme Activation / Q000502:physiology; D066298:In Vitro Techniques; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D009119:Muscle Contraction / Q000187:drug effects; D009388:Neostigmine / Q000494:pharmacology; D009469:Neuromuscular Junction / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D010791:Phrenic Nerve / Q000502:physiology; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011978:Receptors, Nicotinic / Q000187:drug effects*; D019311:Staurosporine; D013569:Synapses / Q000201:enzymology / Q000502:physiology
D000470:Alkaloids; D011978:Receptors, Nicotinic; D009388:Neostigmine; D011493:Protein Kinase C; D019311:Staurosporine; D002118:Calcium