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Regeneration of acetylcholinesterase in clonal neuroblastoma-glioma hybrid NG108-15 cells after soman inhibition: effect of glycyl-l-glutamine
Authors:Jeffrey J. Yourick  Patricia A. Eklo  Michael P. McCluskey  Radharaman Ray
Affiliation:(1) Applied Pharmacology, Branches U.S. Army Medical Research Institute of Chemical Defense Aberdeen Proving Ground, Maryland;(2) U.S. Army Medical Research Institute Chemical Defense, USA;(3) Biochemical Pharmacology, Branches U.S. Army Medical Research Institute of Chemical Defense Aberdeen Proving Ground, Maryland
Abstract:Acetylcholinesterase (AChE) in the clonal NG108-15 cell line has been previously characterized. This cell line represents an in vitro system to study AChE regulation and effects of chemical compounds that may alter AChE activity. Recently, glycyl-L-glutamine (GLG) was demonstrated to function as a neurotrophic factor for maintenance of AChE content in cat denervated superior cervical ganglion cells. In the present study, regeneration of AChE activity in cultures of undifferentiated NG108-15 cells after soman inhibition was investigated in the presence and absence of GLG. Cells were treated with soman (5.5 × 10–6 M) for 15 min and then washed to remove excess soman. Culture medium containing either GLG (10–6, 10–5, or 10–4M) or glycyl-L-glutamic acid (10–6 M) was added to cultures after soman treatment and remained in the medium until cell harvest. Cells were physically detached at various times after soman treatment and specific AChE activity was determined. After soman, AChE activity dramatically decreased to less than 1% of untreated cellular activity at 1 hr. AChE activity gradully increased after 5 hr, while untreated cell AChE activity was regained 20 hr after soman. The t1/2 for AChE regeneration was approximately 10 hr. GLG did not increase the rate of AChE regeneration after soman inhibition. These results indicate that GLG is not a directly acting neurotrophic factor for AChE synthesis in NG108-15 cells after chemical AChE inactivation.Abbreviations AChE acetylcholinesterase - NG108-15 cell neuroblastoma-glioma 108-15 cell - DMEM Dulbecco's modified Eagles minimal essential medium - FBS fetal bovine serum - GLGA glycyl-L-glutamic acid - L-GA L-glutamic acid - GLG glycyl-L-glutamine - GD somanThe opinions or assertions contained herein are the private views of the authors and are not to be construed as reflecting the view of the Department of the Army or the Department of the Army or the Department of Defense.
Keywords:Acetylcholinesterase  NG108-15  Soman  Organophosphorus  Glycyl-L-glutamine  Cell culture
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