title: Morphine-3-glucuronide's neuro-excitatory effects are mediated via indirect activation of N-methyl-D-aspartic acid receptors: mechanistic studies in embryonic cultured hippocampal neurones.
abstract: Indirect evidence indicates that morphine-3-glucuronide (M3G) may contribute significantly to the neuro-excitatory side effects (myoclonus and allodynia) of large-dose systemic morphine. To gain insight into the mechanism underlying M3G's excitatory behaviors, we used fluo-3 fluorescence digital imaging techniques to assess the acute effects of M3G (5-500 microM) on the cytosolic calcium concentration ([Ca(2+)](CYT)) in cultured embryonic hippocampal neurones. Acute (3 min) exposure of neurones to M3G evoked [Ca(2+)](CYT) transients that were typically either (a) transient oscillatory responses characterized by a rapid increase in [Ca(2+)](CYT) oscillation amplitude that was sustained for at least approximately 30 s or (b) a sustained increase in [Ca(2+)](CYT) that slowly recovered to baseline. Naloxone-pretreatment decreased the proportion of M3G-responsive neurones by 10%-25%, implicating a predominantly non-opioidergic mechanism. Although the naloxone-insensitive M3G-induced increases in [Ca(2+)](CYT) were completely blocked by N-methyl-D-aspartic acid (NMDA) antagonists and 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) (alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid/kainate antagonist), CNQX did not block the large increase in [Ca(2+)](CYT) evoked by NMDA (as expected), confirming that M3G indirectly activates the NMDA receptor. Additionally, tetrodotoxin (Na(+) channel blocker), baclofen (gamma-aminobutyric acid(B) agonist), MVIIC (P/Q-type calcium channel blocker), and nifedipine (L-type calcium channel blocker) all abolished M3G-induced increases in [Ca(2+)](CYT), suggesting that M3G may produce its neuro-excitatory effects by modulating neurotransmitter release. However, additional characterization is required.
  
  
  CONCLUSIONS
  Large systemic doses of morphine administered to some patients for cancer pain management have been reported to produce myoclonus and allodynia. Indirect evidence implicates the major morphine metabolite, morphine-3-glucuronide (M3G), in these neuro-excitatory side effects. Hence, this study was designed to gain insight into the cellular mechanism responsible for M3G's neuro-excitatory actions.
authors.affiliation: School of Pharmacy, The University of Queensland, St Lucia Campus, Brisbane, Australia.
authors.forename: Kamondanai
authors.initials: K
authors.lastname: Hemstapat
authors.forename: Gregory R
authors.initials: GR
authors.lastname: Monteith
authors.forename: Deborah
authors.initials: D
authors.lastname: Smith
authors.forename: Maree T
authors.initials: MT
authors.lastname: Smith
mesh_terms: D018750:6-Cyano-7-nitroquinoxaline-2,3-dione / Q000494:pharmacology; D000814:Aniline Compounds; D000818:Animals; D001418:Baclofen / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism; D002121:Calcium Channel Blockers / Q000494:pharmacology; D002478:Cells, Cultured; D000697:Central Nervous System Stimulants / Q000494:pharmacology*; D003600:Cytosol / Q000378:metabolism; D018691:Excitatory Amino Acid Antagonists / Q000494:pharmacology; D005456:Fluorescent Dyes; D018755:GABA Agonists / Q000494:pharmacology; D006624:Hippocampus / Q000187:drug effects / Q000378:metabolism*; D007091:Image Processing, Computer-Assisted; D007150:Immunohistochemistry; D008856:Microscopy, Fluorescence; D009022:Morphine Derivatives / Q000494:pharmacology*; D009270:Naloxone / Q000494:pharmacology; D009292:Narcotic Antagonists / Q000494:pharmacology; D009474:Neurons / Q000187:drug effects / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D016194:Receptors, N-Methyl-D-Aspartate / Q000378:metabolism*; D013779:Tetrodotoxin / Q000494:pharmacology; D014966:Xanthenes
chemical_list: D000814:Aniline Compounds; D002121:Calcium Channel Blockers; D000697:Central Nervous System Stimulants; D018691:Excitatory Amino Acid Antagonists; D005456:Fluorescent Dyes; D018755:GABA Agonists; D009022:Morphine Derivatives; D009292:Narcotic Antagonists; D016194:Receptors, N-Methyl-D-Aspartate; D014966:Xanthenes; C059715:Fluo-3; D009270:Naloxone; D013779:Tetrodotoxin; D018750:6-Cyano-7-nitroquinoxaline-2,3-dione; D001418:Baclofen; C018108:morphine-3-glucuronide; D002118:Calcium