Investigation of the role of intracellular Ca(2+) stores in generation of the muscarinic agonist-induced slow afterdepolarization (sADP) in guinea-pig olfactory cortical neurones in vitro.
1. Intracellular recordings were made from guinea-pig olfactory cortical brain slice neurones to assess the possible role of intracellular Ca(2+) stores in the generation of the slow post-stimulus afterdepolarization (sADP) and its underlying tail current (I(ADP)), induced by muscarinic receptor activation. 2. Caffeine or theophylline (0.5 - 3 mM) reduced the amplitude of the I(ADP) (measured under 'hybrid' voltage clamp) induced in the presence of the muscarinic agonist oxotremorine-M (OXO-M, 10 microM) by up to 96%, without affecting membrane properties or muscarinic depolarization of these neurones. 3. The L-type Ca(2+) channel blocker nifedipine (1, 10 microM) also inhibited I(ADP) (by up to 46%), while ryanodine (10 microM) (a blocker of Ca(2+) release from internal stores) produced a small ( approximately 10%) reduction in I(ADP) amplitude; however, neither 10 microM dantrolene (another internal Ca(2+) release blocker) nor the intracellular Ca(2+) store re-uptake inhibitors thapsigargin (3 microM) or cyclopiazonic acid (CPA, 15 microM) affected I(ADP) amplitude. 4. IBMX (100 microM), a phosphodiesterase inhibitor, also had no effect on I(ADP). Furthermore, inhibition of I(ADP) by caffeine was not reversed by co-application of 100 microM adenosine. 5. Caffeine (3 mM) or nifedipine (10 microM) reduced the duration of presumed Ca(2+) spikes revealed by intracellular Cs(+) loading. When applied in combination, nifedipine and caffeine effects were occlusive, rather than additive, suggesting a common site of action on L-type calcium channels. 6. We conclude that Ca(2+)-induced Ca(2+) release (CICR) from internal stores does not contribute significantly to muscarinic I(ADP) generation in olfactory cortical neurones. However caffeine and theophylline, which enhance CICR in other systems, blocked I(ADP) induction. We suggest that this action might involve a combination of L-type voltage-gated Ca(2+) channel blockade, and a direct inhibitory action on the putative I(ADP) K(+) conductance.
Postlethwaite
M
M
Department of Pharmacology, The School of Pharmacy, 29-39 Brunswick Square, London WC1N 1AX, UK. thwait@cua.ulsop.ac.uk
Constanti
A
A
Libri
V
V
D000200:Action Potentials / Q000187:drug effects*; D000818:Animals; D002110:Caffeine / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism*; D002121:Calcium Channel Blockers / Q000494:pharmacology; D003620:Dantrolene / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004558:Electric Stimulation; D019706:Excitatory Postsynaptic Potentials / Q000187:drug effects; D005260:Female; D006168:Guinea Pigs; D066298:In Vitro Techniques; D007211:Indoles / Q000494:pharmacology; D008297:Male; D008564:Membrane Potentials / Q000187:drug effects; D018721:Muscarinic Agonists / Q000494:pharmacology*; D009474:Neurons / Q000187:drug effects* / Q000502:physiology; D009543:Nifedipine / Q000494:pharmacology; D009833:Olfactory Pathways / Q000166:cytology / Q000187:drug effects* / Q000502:physiology; D010095:Oxotremorine / Q000031:analogs & derivatives / Q000494:pharmacology; D018094:Receptors, Metabotropic Glutamate / Q000187:drug effects / Q000502:physiology; D011976:Receptors, Muscarinic / Q000187:drug effects / Q000502:physiology; D012433:Ryanodine / Q000494:pharmacology; D019837:Ryanodine Receptor Calcium Release Channel / Q000187:drug effects; D009435:Synaptic Transmission / Q000187:drug effects; D019284:Thapsigargin / Q000494:pharmacology
D002121:Calcium Channel Blockers; D007211:Indoles; D018721:Muscarinic Agonists; D018094:Receptors, Metabotropic Glutamate; D011976:Receptors, Muscarinic; D019837:Ryanodine Receptor Calcium Release Channel; D012433:Ryanodine; D002110:Caffeine; D010095:Oxotremorine; C042743:oxotremorine M; D019284:Thapsigargin; D003620:Dantrolene; D009543:Nifedipine; D002118:Calcium; C000543:cyclopiazonic acid