Type of Cell Death and the Role of Acetylcholinesterase Activity in Neurotoxicity Induced by Paraoxon in Cultured Rat Hippocampal Neurons

Abstract

Organophosphate (Ops) neurotoxicity is attributed both to its well-known cholinergic and non-cholinergic effects. In the present study we compared enzymatic and morphologic changes in neurons exposed to paraoxon during one day and one week. The effect of exposure time is important in neurotoxicity of Ops. The longer the exposure time is the more damage is observed in neurons, although there are few investigations about the effect in the post-exposure period. Hippocampal cells were obtained from rat neonates and cultured in Neurobasal/B27. Paraoxon at 50 and 100 μM were added. Inverted microscope and electron microscope were used to study cell morphology and Neutral Red staining was used to measure viability. We also assayed caspase-3 and (acetylcholinesterase) AChE activity. Hoechst staining was utilized to determine the type of cell death. Culture medium was replaced after 24 h in one-day group, however, tests were all carried out at the end of the first week in both group.

The results indicate that paraoxon reduced the viability in a dose-dependent manner. Our results do not confirm apoptosis in either group; it seems that the cell death in one-day exposure group was not AChE dependent. In conclusion, present data imply that the toxicity of paraoxon is both dose and duration dependent, which may even remain after the cessation of exposure.

Abbreviations

AChE:

acetylcholinesterase

CNS:

Central nervous system

CPF:

cholorpyrifos

DTNB:

2,2’-dinitro 5,5’-diptio-dibenzoic acid

ELIZA:

Enzyme linked immunosorbent assay

HEPES:

4-(2-hydroxyethyl)-1 -piperazineethanesulfonic acid

NR:

Neutral Red

NMDA:

N-methyl, D-aspartate

Ops:

organophosphates

SEM:

scanning electron microscopy

VX:

O-ethyl-S-[2(diisopropylamino) ethyl] methylphosphonothiolate

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Acknowledgement

Our great indebtedness to the grant supplier, the Research Center for Chemical Injuries of Baqyatallah University of Medical Science.

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Correspondence to M. Yousefpour.

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Bahrami, F., Yousefpour, M., Mehrani, H. et al. Type of Cell Death and the Role of Acetylcholinesterase Activity in Neurotoxicity Induced by Paraoxon in Cultured Rat Hippocampal Neurons. BIOLOGIA FUTURA 60, 1–13 (2009). https://doi.org/10.1556/ABiol.60.2009.1.1

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Keywords

  • Paraoxon
  • hippocampus
  • cell culture
  • apoptosis
  • cholinesterase activity