Contribution of L-type Ca(2+) channels to evoked transmitter release in cultured Xenopus nerve-muscle synapses.
1. Simultaneous pre- and postsynaptic patch recordings were obtained from the varicosity synapses formed by Xenopus motoneurons on muscle cells in embryonic cultures, in order to elucidate the contribution of N- and L-type Ca(2+) channels to the varicosity Ca(2+) current (I(Ca)) and evoked transmitter release. 2. Although N-type channels are predominant in the varicosities and generally thought to be responsible for all evoked release, in most synapses a fraction of I(Ca) and release could be reversibly blocked by the L-type channel antagonist nifedipine, and enhanced by the agonist Bay K8644. Up to 50 % (mean, 21 %) of the I(Ca) evoked by a voltage clamp waveform mimicking a normal presynaptic action potential (APWF) is composed of L-type current. 3. Surprisingly, the nifedipine-sensitive (L) channels activated more rapidly (time-constant, 0.46 ms at +30 mV) than the nifedipine-insensitive (N) channels (time constant, 1.42 ms). Thus the L-type current would play a disproportionate role in the I(Ca) linked to a normal action potential. 4. The relationship between I(Ca) and release was the same for nifedipine-sensitive and -resistant components. The N- and L-components of I(Ca) are thus equally potent in evoking release. This may represent an immature stage before N-type channels become predominant. 5. Replacing Ca(2+) in the medium with Ba(2+) strongly enhanced the L-type component, suggesting that L-type channels may be inactivated at Ca(2+) levels close to those at rest. 6. We speculate that populations of L-type channels in different parts of the neuron may be recruited or inactivated by fluctuations of the cytosolic Ca(2+) concentration within the physiological range.
Sand
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Department of Biology, University of Oslo, PO Box 1051 Blindern, N-0316 Oslo, Norway.
Chen
B M
BM
Grinnell
A D
AD
D001498:3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester / Q000494:pharmacology; D000109:Acetylcholine / Q000378:metabolism; D000818:Animals; D002120:Calcium Channel Agonists / Q000494:pharmacology; D002121:Calcium Channel Blockers / Q000494:pharmacology; D020746:Calcium Channels, L-Type / Q000378:metabolism*; D020864:Calcium Channels, N-Type / Q000378:metabolism; D002478:Cells, Cultured; D005089:Exocytosis / Q000187:drug effects / Q000502:physiology; D009046:Motor Neurons / Q000166:cytology / Q000502:physiology*; D018485:Muscle Fibers, Skeletal / Q000166:cytology / Q000502:physiology*; D009543:Nifedipine / Q000494:pharmacology; D018408:Patch-Clamp Techniques; D013116:Spinal Cord / Q000166:cytology; D013569:Synapses / Q000378:metabolism; D009435:Synaptic Transmission / Q000187:drug effects / Q000502:physiology*; D014982:Xenopus laevis
D002120:Calcium Channel Agonists; D002121:Calcium Channel Blockers; D020746:Calcium Channels, L-Type; D020864:Calcium Channels, N-Type; D001498:3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester; D009543:Nifedipine; D000109:Acetylcholine