Mechanisms for facilitation of nitric oxide-evoked [3H]GABA release by removal of hydroxyl radical.
We have investigated the mechanisms for enhancement of nitric oxide (NO)-evoked gamma-[3H]aminobutyric acid ([3H]GABA) release from mouse cerebrocortical neurons by hydroxyl radical (.OH) scavengers. .OH scavengers, such as N,N'-dimethylthiourea (DMTU), uric acid, and mannitol, dose-dependently facilitated NO-evoked [3H]GABA release evoked by NO liberated from S-nitroso-N-acetylpenicillamine. Ionomycin-evoked [3H]GABA release, which was significantly inhibited by hemoglobin and an NO synthase, N(G)-methyl-L-arginine, was also enhanced by DMTU. These results indicate that GABA release evoked by both endogenous and exogenous NO is facilitated by .OH scavengers. These enhancing actions of .OH scavengers were completely abolished by Ca2+ removal from incubation buffer and by an L-type voltage-dependent Ca2+ channel (VDCC) inhibitor, nifedipine, whereas each .OH scavenger showed no effects on [3H]GABA release in the absence of NO. Inhibitors for P/Q- and N-type VDCCs had no effects on the enhancement. NO-induced 45Ca2+ influx was also dose-dependently enhanced by .OH scavengers, although 45Ca2+ influx was not altered by .OH scavengers in the absence of NO. Nifedipine abolished this enhancement of the NO-induced 45Ca2+ influx by .OH scavengers. These results indicate that the removal of .OH by its scavengers facilitates the NO-evoked [3H]GABA release dependent on Ca2+ and that this enhancement is due to the increase in Ca2+ influx via L-type VDCCs.
Ohkuma
S
S
Department of Pharmacology, Kawasaki Medical School, Kurashiki, Japan.
Katsura
M
M
Hibino
Y
Y
Hara
A
A
Shirotani
K
K
Ishikawa
E
E
Kuriyama
K
K
D000818:Animals; D002118:Calcium / Q000378:metabolism; D015220:Calcium Channels / Q000187:drug effects; D020746:Calcium Channels, L-Type; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology; D016166:Free Radical Scavengers / Q000494:pharmacology; D017665:Hydroxyl Radical / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D051379:Mice; D009474:Neurons / Q000378:metabolism; D009569:Nitric Oxide / Q000502:physiology*; D013890:Thiourea / Q000031:analogs & derivatives / Q000494:pharmacology; D005680:gamma-Aminobutyric Acid / Q000378:metabolism*
D015220:Calcium Channels; D020746:Calcium Channels, L-Type; D016166:Free Radical Scavengers; D009569:Nitric Oxide; D017665:Hydroxyl Radical; D005680:gamma-Aminobutyric Acid; C038983:1,3-dimethylthiourea; D013890:Thiourea; D002118:Calcium