title: Long-term evaluation of organophosphate toxicity and antidotal therapy in co-cultures of spinal cord and muscle tissue.
abstract: Victims of nerve agents basically require antidotal treatment. There is need for novel antidotes and for therapeutic procedures that are specifically adapted to these patients. To cope with this challenge, in vitro test systems which are easy to handle and allow for conducting long-term studies would be of great benefit. The present work introduces co-cultures of spinal cord and muscle tissue as ex vivo testing systems meeting these criteria. Cell cultures in which functional neuromuscular synapses formed ex vivo were prepared from embryonic mice. Spontaneous muscle activity was recorded by video microscopy. Muscle contractions involved intact neuromuscular transmission as indicated by the effect of succinylcholine, a muscle relaxant that completely abolished muscle activity. At a concentration of 0.75 μM the nerve agent VX reduced the frequency of spontaneous muscle contractions by about 75%. Subsequent application of obidoxime re-established muscle movements. After 24 h of antidotal treatment, muscle activity approached the level of sham-treated cultures and remained stable over the following week. In summary, co-cultures of spinal cord and muscle tissue are promising tools for evaluating the success of antidotal treatment following organophosphate intoxication over a period of at least seven days.
authors.affiliation: Department of Anaesthesiology, Experimental Anaesthesiology Section, Eberhard-Karls-University, Schaffhausenstr. 113, 72072 Tuebingen, Germany.
authors.forename: Berthold
authors.initials: B
authors.lastname: Drexler
authors.forename: Thomas
authors.initials: T
authors.lastname: Seeger
authors.forename: Christian
authors.initials: C
authors.lastname: Grasshoff
authors.forename: Horst
authors.initials: H
authors.lastname: Thiermann
authors.forename: Bernd
authors.initials: B
authors.lastname: Antkowiak
mesh_terms: D000818:Animals; D000931:Antidotes / Q000494:pharmacology* / Q000627:therapeutic use; D018920:Coculture Techniques; D004353:Drug Evaluation, Preclinical / Q000295:instrumentation / Q000379:methods; D004867:Equipment Design; D051379:Mice; D008815:Mice, Inbred Strains; D018715:Microscopy, Video; D009119:Muscle Contraction / Q000187:drug effects; D018485:Muscle Fibers, Skeletal / Q000187:drug effects; D018482:Muscle, Skeletal / Q000187:drug effects* / Q000196:embryology; D009467:Neuromuscular Depolarizing Agents / Q000494:pharmacology; D010755:Organophosphates / Q000633:toxicity*; D009946:Organothiophosphorus Compounds / Q000633:toxicity*; D013116:Spinal Cord / Q000187:drug effects* / Q000196:embryology; D013390:Succinylcholine / Q000494:pharmacology; D013997:Time Factors
chemical_list: D000931:Antidotes; D009467:Neuromuscular Depolarizing Agents; D010755:Organophosphates; D009946:Organothiophosphorus Compounds; D013390:Succinylcholine; C009680:VX-agent