3,4-Diaminopyridine-induced noradrenaline release from CNS tissue as a model for action potential-evoked transmitter release: effects of phorbol ester.
We used rabbit hippocampus slices preincubated with [3H]noradrenaline (NA) and applied short pulses of 3,4-diaminopyridine (3,4-DAP) during superfusion to investigate the mechanism underlying the 3H overflow evoked by 3,4-DAP and the effects of the protein kinase C (PKC) activator, 4 beta-phorbol 12,13-dibutyrate (PDB), in this model. The 3H overflow evoked by 200 microM 3,4-DAP (about 4-5% of tissue-tritium) was largely Ca2+-dependent, tetrodotoxin-sensitive and markedly reduced by clonidine, but it was enhanced by yohimbine. We also demonstrated that the response could be inhibited via presynaptic adenosine (A1-) and opioid (kappa-) receptors. PDB (1 microM) markedly increased the 3,4-DAP-evoked 3H overflow, its effect being almost unchanged following activation of presynaptic alpha 2-, A1- or kappa-receptors. Inhibitors of PKC (polymyxin B, staurosporine) almost abolished the 3,4-DAP-evoked 3H overflow and antagonized the effects of PDB. It is concluded that application of 3,4-DAP (200 microM for 2 min) to brain slices leads to depolarization of the neuronal membrane, Na+ current-carried action potentials, Ca2+ influx and the exocytotic release of NA, which in many aspects resembles the release evoked by electrical field stimulation. The findings with phorbol ester further support the involvement of PKC in transmitter release. Activation of PKC apparently does not directly interfere with signal transduction mechanisms of presynaptic inhibitory receptors on noradrenergic nerve terminals.
Huang
H Y
HY
Institut für Pharmakologie und Toxikologie, Universität Freiburg, F.R.G.
Hertting
G
G
Allgaier
C
C
Jackisch
R
R
D015761:4-Aminopyridine / Q000031:analogs & derivatives* / Q000494:pharmacology; D000200:Action Potentials / Q000187:drug effects; D000077770:Amifampridine; D000818:Animals; D001921:Brain / Q000187:drug effects / Q000378:metabolism*; D004789:Enzyme Activation / Q000187:drug effects; D005071:Evoked Potentials / Q000187:drug effects; D006624:Hippocampus / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism; D066298:In Vitro Techniques; D008954:Models, Biological; D018377:Neurotransmitter Agents / Q000378:metabolism; D009638:Norepinephrine / Q000378:metabolism*; D010703:Phorbol Esters / Q000494:pharmacology*; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism; D011817:Rabbits
D018377:Neurotransmitter Agents; D010703:Phorbol Esters; D015761:4-Aminopyridine; D011493:Protein Kinase C; D000077770:Amifampridine; D009638:Norepinephrine