Nitric oxide augments single Ca(2+) channel currents via cGMP-dependent protein kinase in Kenyon cells isolated from the mushroom body of the cricket brain.
Behavioral and pharmacological studies in insects have suggested that the nitric oxide (NO)/cyclic GMP (cGMP) signaling pathway is involved in the formation of long-term memory (LTM) associated with olfactory learning. However, the target molecules of NO and the downstream signaling pathway are still not known. In this study, we investigated the action of NO on single voltage-dependent Ca(2+) channels in the intrinsic neurons known as Kenyon cells within the mushroom body of the cricket brain, using the cell-attached configuration of the patch-clamp technique. Application of the NO donor S-nitrosoglutathione (GSNO) increased the open probability (NPO) of single Ca(2+) channel currents. This GSNO-induced increase was blocked by ODQ, a soluble guanylate cyclase (sGC) inhibitor, suggesting that the NO generated by GSNO acts via sGC to raise cGMP levels. The membrane-permeable cGMP analog 8-Bro-cGMP also increased the NPO of single Ca(2+) channel currents. Pretreatment of cells with KT5823, a protein kinase G blocker, abolished the excitatory effect of GSNO. These results suggest that NO augments the activity of single Ca(2+) channels via the cGMP/PKG signaling pathway. To gain insight into the physiological role of NO, we examined the effect of GSNO on action potentials of Kenyon cells under current-clamp conditions. Application of GSNO increased the frequency of action potentials elicited by depolarizing current injections, indicating that NO acts as a modulator resulting in a stimulatory signal in Kenyon cells. We discuss the increased Ca(2+) influx through these Ca(2+) channels via the NO/cGMP signaling cascade in relation to the formation of olfactory LTM.
Tokyo Gakugei University Senior High School, Tokyo 154-0002, Japan.
Kumiko
K
Kosakai
Department of Biology, Tokyo Gakugei University, Tokyo 184-8501, Japan.
Yuuki
Y
Tsujiuchi
Department of Biology, Tokyo Gakugei University, Tokyo 184-8501, Japan. Electronic address: myoshi@u-gakugei.ac.jp.
Masami
M
Yoshino
D000200:Action Potentials; D000818:Animals; D001921:Brain / Q000166:cytology / Q000378:metabolism; D015220:Calcium Channels / Q000502:physiology*; D002227:Carbazoles / Q000494:pharmacology; D006152:Cyclic GMP / Q000031:analogs & derivatives / Q000494:pharmacology; D017869:Cyclic GMP-Dependent Protein Kinases / Q000037:antagonists & inhibitors / Q000502:physiology*; D006135:Gryllidae / Q000166:cytology / Q000378:metabolism*; D008297:Male; D024521:Mushroom Bodies / Q000166:cytology / Q000378:metabolism*; D009474:Neurons / Q000187:drug effects / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism*; D020030:Nitric Oxide Donors / Q000494:pharmacology; D010069:Oxadiazoles / Q000494:pharmacology; D011810:Quinoxalines / Q000494:pharmacology; D026422:S-Nitrosoglutathione / Q000494:pharmacology; D015398:Signal Transduction
C095284:1H-(1,2,4)oxadiazolo(4,3-a)quinoxalin-1-one; D015220:Calcium Channels; D002227:Carbazoles; D020030:Nitric Oxide Donors; D010069:Oxadiazoles; D011810:Quinoxalines; C073601:KT 5823; C016276:8-bromocyclic GMP; D009569:Nitric Oxide; D026422:S-Nitrosoglutathione; D017869:Cyclic GMP-Dependent Protein Kinases; D006152:Cyclic GMP
Cricket; Kenyon cell; L-type Ca(2+) channel; Mushroom body; Nitric oxide