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Regeneration of acetylcholinesterase in clonal neuroblastoma-glioma hybrid NG108-15 cells after soman inhibition: effect of glycyl-l-glutamine

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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.

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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:

soman

References

  • BROGDON, W.G and DICKINSON, C.M. (1983). “A microassay system for measuring esterase activity and protein concentration in small samples and in high-pressure liquid chromatography eluate fractions.” Anal. Biochem. 131:499–503.

    Google Scholar 

  • ELLMAN, G.L., COURTNEY, K.D., ANDRES, V., and FEATHERSTONE, R.M. (1961). “A new and colorimetric determination of acetylcholinesterase activity.” Biochem. Pharmacol. 7:88–95.

    Google Scholar 

  • HAMPRECHT, B. (1977). “Structural, electrophysiological, biochemical, and pharmacological properties of neuroblastoma-glioma cell hybrids in cell culture.” Int. Rev. Cytol. 49:99–170.

    Google Scholar 

  • HAYNES, L.W. and SMITH, M.E. (1985). “Induction of endplate-specific acetylcholinesterase by Β-endorphin C-terminal dipeptide in rat and chick muscle cells. rd Biochem. Soc. Trans. 13:174–175.

    Google Scholar 

  • KOELLE, G.B., SANVILLE, U.J., RICKARD, K.K., and WILLIAMS, J.E. (1984). “Partial characterization of the neurotrophic factor for maintenance of actylcholinesterase and butyrylcholinesterase in the preganglionically denervated superior cervical ganglion of the cat in vivo.” Proc. Natl. Acad. Sci. USA 81:6539–6542.

    Google Scholar 

  • KOELLE, G.B., SANVILLE, U.J., and WALL, S.J. (1985). “Glycyl-L-glutamine, a precursor, and glycyl-L-glutamic acid, a neuroophic factor for maintenance of acetylcholinesterase and butyrylcholinesterase in the preganglionically denervated superior cervical ganglion of the cat in vivo.” Proc. Natl. Acad. Sci. USA 82:5213–5217.

    Google Scholar 

  • KOELLE, G.B., SANVILLE, U.J., THAMPI, N.S., and WALL, S.J. (1986). “L-Glumatic acid, a neurotrophic factor for maintenance of acetylcholinesterase and butyrylcholinestarase in the preganglionically denervated superior cervical ganglion of the cat.“ Proc. Nat. Acad. Sci. USA 83:2751–2754.

    Google Scholar 

  • KOELLE, G.B., SANVILLE, U.J., and THAMPI, N.S. (1987). “Direct neurotrophic action of glycyl-L-glutamine in the maintenance of acetylcholinesterase and butyrylcholinesterase in the preganglionically denervated superior cervical ganglion of the cat.” Proc. Natl. Acad. Sci. USA 84:6944–6947.

    Google Scholar 

  • KOELLE, G.B. (1988). “Enhancement of acetylcholinesterase synthesis by glycyl-L-glutamate: an example of a small peptide that regulates differential transcription?.” Trends in Pharm. Sci. 9:318–321.

    Google Scholar 

  • KOELLE, G.B., MASSOULIÉ, J., EUGENE, D., and MELONE, M.A.B. (1988). “Effects of glycyl-L-glutamine in vitro on the molecular forms of acetylcholinesterase in the preganglionically denervated superior cervical ganglion of the cat.” Proc. Natl. Acad. Sci. USA 85:1686–1690.

    Google Scholar 

  • KOELLE, G.B. and HAN, M.S. (1989). “Effect of glycyl-L-glutamine on the rate of regeneration of acetylcholinesterase in the rat gastrocnemius muscle after diisopropyl phosphorofluoridate administration.” Proc. Natl. Acad. Sci. USA 86: 4331–4333.

    Google Scholar 

  • KOELLE, G.B., O'NEILL, J.J., THAMPI, N.S., and HAN, M.S. (1989). “Glycyl-L-glutamine opposes the fall in choline acetyltransferase in the denervated superior cervical ganglion of the cat.” Proc. Natl. Acad. Sci. USA 86:10153–10155.

    Google Scholar 

  • LANKS, K.W., DORWIN, J.M., and PAPIRMEISTER, B. (1974). “Increased rate of a acetylcholinesterase synthesis in differentiating neuroblastoma cells.” J. Cell Biol. 63:824–830.

    Google Scholar 

  • LOWRY, O.H., ROSEBROUGH, N.J., FARR, A.L., and RANDALL, R.J. (1951). “Protein measurement with the folin phenol reagent.” J. Biol. Chem. 193:265–275.

    Google Scholar 

  • MCGEE, R., SIMPSON, P., CHRISTIAN, C., MATA, M., NELSON, P., and NIRENBERG, M. (1978). “Regulation of acetylcholinerelease from neuroblastoma × glioma hybrid cells.” Proc. Natl. Acad. Sci. USA 75:1314–1318.

    Google Scholar 

  • NIRENBERG, M., WILSON, S.P., HIGASHIDA, H., ROTTER, A., KREUGER, K., BUSIS, N., RAY, R., KENIMER, J., ADLER, M., and FUKUI, H. (1983). “Synapse formation by neuroblastoma hybrid cells.” Cold spring Harbor Symp. Quant. Biol 48:707–715.

    Google Scholar 

  • RAY, R., KELLY, R.A., MURRAY, C.E., and STORM, C.R. (1984).“ Changes in acetylcholinesterase activity and molecular forms in a clonal neuroblastoma-glioma hybrid NG108–15 cell line during differentiation.” Soc. Neurosci. Abstr. 10:1038.

    Google Scholar 

  • RAY, R. and CLARK, O.E. (1988). “A computer-assisted automated colorimetric microassay system for determination of enzyme activity, kinetics, and concentration-response relationship.” FASEB. J. 2:A1541.

    Google Scholar 

  • RIEGER, F., FAIVRE-BAUMAN, A., BENDA, P., and VIGNY, M. (1976). “Molecular forms of acetylcholinesterase: Their de novo synthesis in mouse neuroblastoma cells.” J. Neurochem. 27:1059–1063.

    Google Scholar 

  • TAYLOR, P. (1990). “Anticholinesterase agents.”In: The Pharmacological Basis of Therapeutics (A.E. Gilman, T.W. Rall, A.S. Nies, and P. Taylor, eds.), 8th edition, pp. 131–149. Pergamon Press, New York.

    Google Scholar 

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The 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.

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Yourick, J.J., Eklo, P.A., McCluskey, M.P. et al. Regeneration of acetylcholinesterase in clonal neuroblastoma-glioma hybrid NG108-15 cells after soman inhibition: effect of glycyl-l-glutamine. Cell Biol Toxicol 7, 229–237 (1991). https://doi.org/10.1007/BF00250977

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