Muscarinic receptor-mediated dual regulation of ADP-ribosyl cyclase in NG108-15 neuronal cell membranes

Haruhiro Higashida, Shigeru Yokoyama, Minako Hashii, Megumi Taketo, Masaharu Higashida, Tatsunori Takayasu, Tohru Ohshima, Shin Takasawa, Hiroshi Okamoto, Mami Noda

Research output: Contribution to journalArticlepeer-review

82 Citations (Scopus)


Cyclic ADP-ribose (cADP-ribose) is an endogenous modulator of ryanodine- sensitive Ca2+ release channels. An unsolved question is whether or not cADP-ribose mediates intracellular signals from hormone or neurotransmitter receptors. The first step in this study was to develop a TLC method to measure ADP-ribosyl cyclase, by which conversion of [3H]NAD+ to [3H]cADP- ribose was confirmed in COS-7 cells overexpressing human CD38. A membrane fraction of NG108-15 neuroblastoma x glioma hybrid cells possessed ADP- ribosyl cyclase activity measured by TLC. Carbamylcholine increased this activity by 2.6-fold in NG108-15 cells overexpressing m1 or m3 muscarinic acetylcholine receptors (mAChRs), but inhibited it by 30-52% in cells expressing m2 and/or m4 mAChRs. Both of these effects were mimicked by GTP. Pretreatment of cells with cholera toxin blocked the activation, whereas pertussis toxin blocked the inhibition. Application of carbamylcholine caused significant decreases in NAD+ concentrations in untreated m1-transformed NG108-15 cells, but an increase in cholera toxin-treated cells. These results suggest that mAChRs couple to ADP-ribosyl cyclase within cell membranes via trimeric G proteins and can thereby control cellular function by regulating cADP-ribose formation.

Original languageEnglish
Pages (from-to)31272-31277
Number of pages6
JournalJournal of Biological Chemistry
Issue number50
Publication statusPublished - 1997 Dec 12


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