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Cerebral acetylcholine and choline contents and turnover following low-dose acetylcholinesterase inhibitors treatment in rats

  • Organ Toxicity and Mechanisms
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Abstract

Male Sprague-Dawley rats were treated for 3 weeks with (1) regular tap drinking water plus subcutaneous (s.c.) saline (0.5 ml/kg) injections three times/week, (2) pyridostigmine bromide (PB) in drinking water (80 mg/L) plus s.c. saline injections three times/week, (3) regular tap drinking water plus s.c. sarin (0.5 × LD50) injections three times/week, or (4) PB in drinking water plus s.c. sarin injections three times/week. Repeated doses of sarin, in the presence or absence of PB, were devoid of acute toxicity during the three-week treatment period. Two, 4, and 16 weeks post-treatment, animals were given an intravenous pulse injection of choline labeled with 4 deuterium atoms (D4Ch) followed, after 1 min, by microwave fixation of the brain in vivo. Tissue levels of endogenous acetylcholine (D0ACh), endogenous choline (D0Ch), D4Ch, and ACh synthesized from D4Ch (D4ACh) were measured by gas-chromatography mass-spectrometry in hippocampus, infundibulum, mesencephalon, neocortex, piriform cortex, and striatum. Ch uptake from blood and ACh turnover were estimated from D4Ch and D4ACh concentrations in brain tissue, respectively. Statistically significant differences among brain regions were found for D0Ch, D4Ch, D0ACh and D4ACh at 2, 4 and 16 weeks post-treatment. However, differences in the values of these parameters between control and drug treatments were found only for D0ACh and D0Ch at 2 and 4 weeks, but not at 16 weeks post-treatment. In conclusion, the results from these experiments do not support a delayed or persistent alteration in cholinergic function after exposure to low doses of PB and/or sarin.

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Abbreviations

AChE:

Acetylcholinesterase

CNS:

Central nervous system

D0ACh:

Endogenous acetylcholine

D0Ch:

Endogenous choline

D4ACh:

Acetylcholine labeled with 4 deuterium atoms

D9ACh:

Acetylcholine labeled with 9 deuterium atoms

D4Ch:

Choline labeled with 4 deuterium atoms

D9Ch:

Choline labeled with 9 deuterium atoms

DFP:

Diisopropylfluorophosphate

GCMS:

Gas-chromatograph mass-spectrometry

OP:

Organophosphorus compound

PB:

Pyridostigmine bromide

PGW:

Persian Gulf War

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Acknowledgments

The authors express their great appreciation to Mrs. Tami Rowland and Mr. Harlan Shafer for their excellent technical assistance. This work was supported by the US Army Medical Research and Materiel Command under Contract Order DAMD17-00 200015.Research was conducted in compliance with the Animal Welfare Act and other Federal statutes and regulations relating to animals and experiments involving animals and adheres to principles stated in the Guide for the Care and Use of Laboratory Animals, National Research Council, 1996. The facilities where this research was conducted are fully accredited by the Association for Assessment and Accreditation of Laboratory Animal Care International. The opinions or assertions contained herein are the private views of the authors, and are not to be construed as reflecting the views of the Department of the Army or the Department of Defense.

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Authors and Affiliations

  1. Research Division, US Army Medical Research Institute of Chemical Defense, Aberdeen Proving Ground, MD, 21010-5400, USA

    Tsung-Ming Shih

  2. Research Department, VA GLA Healthcare System, Los Angeles, CA, USA

    Oscar U. Scremin, Margareth Roch, Ly Huynh & Wei Sun

  3. Physiology Department, David Geffen School of Medicine at UCLA, Los Angeles, CA, USA

    Oscar U. Scremin

  4. Medical and Molecular Pharmacology Department, David Geffen School of Medicine at UCLA, Los Angeles, CA, USA

    Donald J. Jenden

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  1. Tsung-Ming Shih
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  2. Oscar U. Scremin
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  3. Margareth Roch
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  4. Ly Huynh
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  5. Wei Sun
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  6. Donald J. Jenden
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Correspondence to Tsung-Ming Shih.

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Shih, TM., Scremin, O.U., Roch, M. et al. Cerebral acetylcholine and choline contents and turnover following low-dose acetylcholinesterase inhibitors treatment in rats. Arch Toxicol 80, 761–767 (2006). https://doi.org/10.1007/s00204-006-0101-5

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  • DOI: https://doi.org/10.1007/s00204-006-0101-5

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