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Related Experiment Videos

Brain fixation for acetylcholine measurements

N Bertrand1, P Beley, A Beley

  • 1Laboratoire de Pharmacodynamie, Faculté de Pharmacie, Dijon, France.

Journal of Neuroscience Methods
|July 1, 1994
PubMed
Summary

Microwave irradiation of rodent heads effectively inactivates brain enzymes, preserving acetylcholine levels for accurate neurochemical studies. This simple method offers a valuable approach for brain fixation in small animal research.

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Area of Science:

  • Neuroscience
  • Biochemistry
  • Analytical Chemistry

Background:

  • Post-mortem degradation of neurotransmitters like acetylcholine (ACh) complicates neurochemical analysis.
  • Accurate measurement of brain ACh and choline (Ch) requires rapid inactivation of enzymes responsible for their metabolism.

Purpose of the Study:

  • To evaluate microwave irradiation of rodent heads as a rapid brain fixation method.
  • To assess the efficacy of microwave fixation in preventing post-mortem changes in brain ACh and Ch levels.
  • To validate the method for neurochemical studies in small animal models.

Main Methods:

  • Utilized a 750 W microwave oven for brain fixation in decapitated rats and mice.
  • Irradiation times were optimized: 5.5 seconds for rat heads and 3 seconds for mouse heads.

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  • Quantified acetylcholine and choline content in specific brain regions (striatum, hippocampus, cortex) using established analytical techniques.

    • Main Results:

    • Complete inactivation of acetylcholinesterase (AChE) and choline acetyltransferase (ChAT) was achieved.
    • Measured ACh and Ch levels in rat brain regions (striatum, hippocampus, cortex) were consistent with literature values.
    • Measured ACh and Ch levels in mouse brain regions (striatum, hippocampus, cortex) were consistent with literature values.

    Conclusions:

    • Microwave irradiation of severed rodent heads is a simple and effective method for brain fixation.
    • This technique successfully preserves endogenous acetylcholine and choline levels by inactivating key metabolic enzymes.
    • The described procedure is a valuable tool for neurochemical research on brain acetylcholine in small animals.