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Acta Biologica Hungarica

, Volume 60, Issue 1, pp 1–13 | Cite as

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

  • F. Bahrami
  • M. YousefpourEmail author
  • H. Mehrani
  • L. Golmanesh
  • S. H. Sadraee
  • A. Khoshbaten
  • A. Asgari
Article

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.

Keywords

Paraoxon hippocampus cell culture apoptosis cholinesterase activity 

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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Notes

Acknowledgement

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

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© Akadémiai Kiadó, Budapest 2009

This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.

Authors and Affiliations

  • F. Bahrami
    • 1
  • M. Yousefpour
    • 1
    Email author
  • H. Mehrani
    • 2
  • L. Golmanesh
    • 3
  • S. H. Sadraee
    • 4
  • A. Khoshbaten
    • 1
    • 2
  • A. Asgari
    • 1
    • 2
  1. 1.Department of Physiology and BiophysicsBaqyatallah University of Medical ScienceTehranIran
  2. 2.Research Center for Chemical InjuriesBaqyatallah University of Medical ScienceTehranIran
  3. 3.Research Center for Molecular BiologyBaqyatallah University of Medical ScienceTehranIran
  4. 4.Department of Anatomy and HistologyBaqyatallah University of Medical ScienceTehranIran

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