Dynorphins directly inhibit neuronal nicotinic acetylcholine receptors in PC12 cells.
The authors have previously reported that dynorphin A (1-17), an endogenous kappa opioid agonist, inhibits the current mediated through neuronal nicotinic acetylcholine receptors (nAChRs) without the involvement of opioid receptors or G-proteins. We have further characterized this action to elucidate the mechanisms. The nicotine-induced current was studied in PC12 cells using patch-clamp techniques. In the whole-cell configuration, four kinds of dynorphins with different lengths, dynorphin A (1-17) (1-13) (2-13) and (1-8), similarly inhibited the nicotine-induced inward current at 1 microM and accelerated the current decay. The inhibition by dynorphin A (1-17) was not antagonized by the increasing concentrations of nicotine. The current-voltage relationship revealed that dynorphin's inhibition was voltage independent at the membrane potentials from -30 to -70 mV. The inhibition was not affected by pretreatment with pertussis toxin (PTX) or inclusion of staurosporine into the pipette solution. The inhibitory effect of dynorphin A (1-17) was well preserved in the outside-out patch configuration. Analysis of the nicotine-induced noise and single-channel kinetics revealed that dynorphin A(1-17) reduced open time without changing the amplitude of the unitary current. We found that the inhibitory effect on neuronal nAChRs is shared by all four dynorphins studied. The inhibition appears to be non-competitive and voltage independent. The outside-out recording together with other experiments indicated that a major part of this inhibition is not mediated through cytoplasmic messengers, but based on the direct action of dynorphins on neuronal nAChRs leading to the reduction of open time.
Itoh
H
H
Department of Anaesthesiology, Yokohama City University School of Medicine, Yokohama, Japan.
Andoh
T
T
Watanabe
I
I
Sasaki
T
T
Kamiya
Y
Y
Okumura
F
F
D000818:Animals; D004305:Dose-Response Relationship, Drug; D004399:Dynorphins / Q000494:pharmacology*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D015640:Ion Channel Gating / Q000187:drug effects; D008564:Membrane Potentials / Q000187:drug effects; D020932:Nerve Growth Factor / Q000494:pharmacology; D009433:Neural Inhibition / Q000187:drug effects; D009474:Neurons / Q000737:chemistry / Q000502:physiology*; D009538:Nicotine / Q000494:pharmacology; D018722:Nicotinic Agonists / Q000494:pharmacology; D018733:Nicotinic Antagonists / Q000494:pharmacology*; D016716:PC12 Cells; D018408:Patch-Clamp Techniques; D010446:Peptide Fragments / Q000494:pharmacology; D037342:Pertussis Toxin; D051381:Rats; D011978:Receptors, Nicotinic / Q000502:physiology*; D019311:Staurosporine / Q000494:pharmacology; D010566:Virulence Factors, Bordetella / Q000494:pharmacology
D004791:Enzyme Inhibitors; D018722:Nicotinic Agonists; D018733:Nicotinic Antagonists; D010446:Peptide Fragments; D011978:Receptors, Nicotinic; D010566:Virulence Factors, Bordetella; D009538:Nicotine; C026843:dynorphin (1-13); D004399:Dynorphins; C026770:dynorphin (1-8); C036730:dynorphin A, desTyr(1)-desTrp(14)-desAsp(15)-desAsn(16)-desGlu(17)-; D020932:Nerve Growth Factor; D037342:Pertussis Toxin; D019311:Staurosporine