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Active and peripheral anionic sites of acetylcholinesterase have differential modulation effects on cell proliferation, adhesion and neuritogenesis in the NG108-15 cell line.
Toxicol Lett. 2014 Oct 15; 230(2):122-31.TL

Abstract

The classical enzymatic role of acetylcholinesterase (AChE) is to terminate impulse transmission at cholinergic synapses through rapid hydrolysis of acetylcholine (ACh). Inactivation of this enzyme's catalytic site is the primary mechanism of acute toxicity of OP insecticides (e.g. parathion, chlorpyrifos). There is now sufficient evidence to suggest that AChE has a neurotrophic function that may be altered by organophosphate (OP) exposure, resulting in defects of neuronal growth and development, though the clarification of the mechanisms involved require further in vitro investigation. In the present study, the mouse neuroblastoma×rat glioma hybrid NG108-15 cell line was used to investigate the differential effects between inhibition of the catalytic site and peripheral anionic site (PAS) of acetylcholinesterase (AChE) on cell adhesion, proliferation and neuritogenesis, in the presence and absence of human red blood cell (hRBC) AChE (ED3.1.1.7). AChE active-site inhibitor paraoxon (PO; 0.1-1.0μM), when added to NG108-15 cells grown on AChE-coated plates, had no effect on cell proliferation, but exerted a significant reduction in strongly adherent viable cells accompanied by mostly short process formations, with 18% of cells considered to be neuritogenic, similar to that observed on uncoated plates. In contrast, PO had no significant effect on cell adhesion and proliferation of NG108-15 cells on uncoated plates. The PAS-ligand thioflavin-T (Th-T; 0.5-25μM), however, decreased cell adhesion and proliferation, on both uncoated and ACh-E coated plates, with less magnitude on AChE-coated plates. Taken together, these results suggest that strong cell adherence and neuritogenesis are sensitive to PO in this cell culture model, with no impact on proliferation, in the presence of membrane bound AChE-coating, while there is no sensitivity to PO on uncoated plates. On the other hand, binding of Th-T directly to the PAS affects both cell adherence and proliferation, with less magnitude in the presence of membrane-bound AChE. The current study indicates that PO is deleterious in neural development during critical periods of strong cell adhesion and differentiation, interfering with AChE trophic function.

Authors+Show Affiliations

Rutgers, New Jersey Medical School-Graduate School of Biomedical Sciences, 185 South Orange Avenue, MSB H609, Newark, NJ 07103, United States. Electronic address: hcampanha@ymail.com.REQUIMTE, Laboratory of Toxicology, Department of Biological Sciences, Faculty of Pharmacy, University of Porto, Rua de Jorge Viterbo Ferreira no. 228, 4050-313 Porto, Portugal.U.S. Environmental Protection Agency, National Center for Environmental Assessment, Washington, DC, United States.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

24680925

Citation

Campanha, Helen M., et al. "Active and Peripheral Anionic Sites of Acetylcholinesterase Have Differential Modulation Effects On Cell Proliferation, Adhesion and Neuritogenesis in the NG108-15 Cell Line." Toxicology Letters, vol. 230, no. 2, 2014, pp. 122-31.
Campanha HM, Carvalho F, Schlosser PM. Active and peripheral anionic sites of acetylcholinesterase have differential modulation effects on cell proliferation, adhesion and neuritogenesis in the NG108-15 cell line. Toxicol Lett. 2014;230(2):122-31.
Campanha, H. M., Carvalho, F., & Schlosser, P. M. (2014). Active and peripheral anionic sites of acetylcholinesterase have differential modulation effects on cell proliferation, adhesion and neuritogenesis in the NG108-15 cell line. Toxicology Letters, 230(2), 122-31. https://doi.org/10.1016/j.toxlet.2014.03.012
Campanha HM, Carvalho F, Schlosser PM. Active and Peripheral Anionic Sites of Acetylcholinesterase Have Differential Modulation Effects On Cell Proliferation, Adhesion and Neuritogenesis in the NG108-15 Cell Line. Toxicol Lett. 2014 Oct 15;230(2):122-31. PubMed PMID: 24680925.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Active and peripheral anionic sites of acetylcholinesterase have differential modulation effects on cell proliferation, adhesion and neuritogenesis in the NG108-15 cell line. AU - Campanha,Helen M, AU - Carvalho,Félix, AU - Schlosser,Paul M, Y1 - 2014/03/27/ PY - 2013/09/26/received PY - 2014/03/02/revised PY - 2014/03/19/accepted PY - 2014/4/1/entrez PY - 2014/4/1/pubmed PY - 2014/12/15/medline KW - Acetylcholinesterase KW - Acylation site KW - Cell adhesion KW - Neuritogenesis KW - Peripheral anionic site KW - Proliferation SP - 122 EP - 31 JF - Toxicology letters JO - Toxicol Lett VL - 230 IS - 2 N2 - The classical enzymatic role of acetylcholinesterase (AChE) is to terminate impulse transmission at cholinergic synapses through rapid hydrolysis of acetylcholine (ACh). Inactivation of this enzyme's catalytic site is the primary mechanism of acute toxicity of OP insecticides (e.g. parathion, chlorpyrifos). There is now sufficient evidence to suggest that AChE has a neurotrophic function that may be altered by organophosphate (OP) exposure, resulting in defects of neuronal growth and development, though the clarification of the mechanisms involved require further in vitro investigation. In the present study, the mouse neuroblastoma×rat glioma hybrid NG108-15 cell line was used to investigate the differential effects between inhibition of the catalytic site and peripheral anionic site (PAS) of acetylcholinesterase (AChE) on cell adhesion, proliferation and neuritogenesis, in the presence and absence of human red blood cell (hRBC) AChE (ED3.1.1.7). AChE active-site inhibitor paraoxon (PO; 0.1-1.0μM), when added to NG108-15 cells grown on AChE-coated plates, had no effect on cell proliferation, but exerted a significant reduction in strongly adherent viable cells accompanied by mostly short process formations, with 18% of cells considered to be neuritogenic, similar to that observed on uncoated plates. In contrast, PO had no significant effect on cell adhesion and proliferation of NG108-15 cells on uncoated plates. The PAS-ligand thioflavin-T (Th-T; 0.5-25μM), however, decreased cell adhesion and proliferation, on both uncoated and ACh-E coated plates, with less magnitude on AChE-coated plates. Taken together, these results suggest that strong cell adherence and neuritogenesis are sensitive to PO in this cell culture model, with no impact on proliferation, in the presence of membrane bound AChE-coating, while there is no sensitivity to PO on uncoated plates. On the other hand, binding of Th-T directly to the PAS affects both cell adherence and proliferation, with less magnitude in the presence of membrane-bound AChE. The current study indicates that PO is deleterious in neural development during critical periods of strong cell adhesion and differentiation, interfering with AChE trophic function. SN - 1879-3169 UR - https://www.unboundmedicine.com/medline/citation/24680925/Active_and_peripheral_anionic_sites_of_acetylcholinesterase_have_differential_modulation_effects_on_cell_proliferation_adhesion_and_neuritogenesis_in_the_NG108_15_cell_line_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0378-4274(14)00134-9 DB - PRIME DP - Unbound Medicine ER -