Inhibition of norepinephrine and acetylcholine release from human neocortex by omega-conotoxin GVIA.
Superfused slices of human neocortex, prepared from surgically removed tissue (to gain access to subcortical tumors) and prelabeled selectively with [3H]norepinephrine (NE) or [3H]choline, were stimulated electrically to evoke tritium overflow. This tritium overflow was abolished by the sodium channel blocker tetrodotoxin and by withdrawal of extracellular Ca++. Thus, the action potential-induced, exocytotic tritium overflow supports the assumption of a quasiphysiological release of NE from noradrenergic and of acetylcholine (ACh) from cholinergic nerve terminals, respectively. In addition, the modulation of NE release by adrenoceptor ligands displayed the appropriate pharmacology of alpha-2 autoreceptors; ACh release was modulated by muscarinic ligands. Both NE and ACh release decreased with the age of the patients. The effects of drugs on NE and ACh release were not age-related. The peptide modulator of the N-type voltage sensitive Ca++ channel, omega-conotoxin GVIA, inhibited NE release with an IC50 of about 14 nM and ACh release with an IC50 of about 3 nM, whereas L-type modulators were ineffective. The binding of [125I]omega-conotoxin GVIA to human neocortical membranes was of high affinity (KD = 1.3 pM) to one site (nH = 0.97) of substantial density (maximum binding = 878 fmol/mg of protein); the binding of the L-type modulator [3H]isradipine to these membranes was also of high affinity (KD = 89 pM) to one site (nH = 1.03) of lesser density (maximum binding = 429 fmol/mg of protein). In conclusion, Ca++ entry through N-type Ca++ channels, rather then L-type Ca++ channels, predominates in subserving NE and ACh release from noradrenergic and cholinergic nerve terminals, respectively, of human neocortex.
Feuerstein
T J
TJ
Neurologische Universitätsklinik, Freiburg, Federal Republic of Germany.
Dooley
D J
DJ
Seeger
W
W
D000109:Acetylcholine / Q000378:metabolism*; D000293:Adolescent; D000328:Adult; D000367:Age Factors; D000368:Aged; D001665:Binding Sites; D015220:Calcium Channels / Q000502:physiology; D002540:Cerebral Cortex / Q000378:metabolism*; D002648:Child; D002675:Child, Preschool; D006801:Humans; D017275:Isradipine; D008875:Middle Aged; D008978:Mollusk Venoms / Q000378:metabolism / Q000494:pharmacology*; D009638:Norepinephrine / Q000378:metabolism*; D011725:Pyridines / Q000378:metabolism; D013268:Stimulation, Chemical; D013779:Tetrodotoxin / Q000494:pharmacology; D020866:omega-Conotoxin GVIA
D015220:Calcium Channels; D008978:Mollusk Venoms; D011725:Pyridines; D013779:Tetrodotoxin; D020866:omega-Conotoxin GVIA; D000109:Acetylcholine; D009638:Norepinephrine; D017275:Isradipine