The effects of presynaptic calcium channel modulation by roscovitine on transmitter release at the adult frog neuromuscular junction.
Calcium (Ca2+) influx through presynaptic calcium channels triggers transmitter release, and any alterations in the gating of these calcium channels results in changes in the magnitude of transmitter released. We used (R)-roscovitine, a cyclin-dependent kinase inhibitor that also appears to act directly on calcium channels, as a tool to modulate presynaptic calcium influx and study effects on transmitter release. We show that this compound increased the quantal content of acetylcholine released from the Rana frog motor nerve terminal (by 149%) without changing paired-pulse facilitation (under low calcium conditions). In contrast, exposure to 3,4-diaminopyridine (DAP; which similarly affects transmitter release by partially blocking potassium channels, altering the shape of the presynaptic action potential, and indirectly increasing calcium entry) increased paired-pulse facilitation (by 23%). In addition, we show that (R)-roscovitine predominately slowed deactivation kinetics of calcium current (by 427%) recorded from Xenopus frog motoneurons, and as a result, increased the integral of calcium channel current evoked by a physiological action potential waveform (by 44%). Because we did not observe any significant effects of structurally related cyclin-dependent kinase inhibitors [(S)-roscovitine or olomoucine] on evoked transmitter release or calcium current kinetics, it appears that these effects of (R)-roscovitine are independent of cyclin-dependent kinases (cdks). In summary, we hypothesize that (R)-roscovitine effects on transmitter release at the adult frog neuromuscular junction (NMJ) are mediated by its effects on calcium channel gating, and these effects increase our understanding of calcium triggered secretion at this synapse.
Cho
Soyoun
S
Department of Neuroscience, Center for Neuroscience, 446 Crawford Hall, University of Pittsburgh, Pittsburgh, PA 15260, USA.
Meriney
Stephen D
SD
D015761:4-Aminopyridine / Q000031:analogs & derivatives / Q000494:pharmacology; D000077770:Amifampridine; D000818:Animals; D015220:Calcium Channels / Q000378:metabolism*; D019706:Excitatory Postsynaptic Potentials / Q000187:drug effects; D007701:Kinetin / Q000494:pharmacology; D009469:Neuromuscular Junction / Q000187:drug effects / Q000502:physiology; D018408:Patch-Clamp Techniques; D026902:Potassium Channel Blockers / Q000494:pharmacology; D017729:Presynaptic Terminals / Q000502:physiology*; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D011687:Purines / Q000494:pharmacology*; D011894:Rana pipiens; D000077546:Roscovitine; D009435:Synaptic Transmission / Q000187:drug effects / Q000502:physiology; D014982:Xenopus laevis
D015220:Calcium Channels; D026902:Potassium Channel Blockers; D047428:Protein Kinase Inhibitors; D011687:Purines; D000077546:Roscovitine; C090046:olomoucine; D015761:4-Aminopyridine; D007701:Kinetin; D000077770:Amifampridine