Mass Spectrometric Enzyme Histochemistry for Choline Acetyltransferase Reveals de Novo Acetylcholine Synthesis in Rodent Brain and Spinal Cord

Emi Takeo, Yuki Sugiura, Yuichiro Ohnishi, Haruhiko Kishima, Eiichiro Fukusaki, Shuichi Shimma

Research output: Contribution to journalArticlepeer-review

Abstract

Choline acetyltransferase (ChAT), responsible for the synthesis of acetylcholine, plays an important role in neurotransmission. However, no method to visualize the ChAT activity in tissues has been reported to date. In this study, mass spectrometry imaging (MSI) was used to visualize ChAT activity in situ, which is difficult with conventional enzyme histochemistry. By using choline chloride-trimethyl-d9 (choline-d9) as a substrate and simultaneously supplying an inhibitor of cholinesterase to tissues, we succeeded in directly visualizing the ChAT activity in the rodent brain and spinal cord. The findings revealed heterogeneous ChAT activity in the striatum of the mouse brain and in the spinal lower motor neurons that connect the anterior horn to the ventral root. Furthermore, extending the developed method to spinal cord injury (SCI) model mice revealed the site-specific effect of primary and secondary injury on ChAT activity. This study shows that the MSI-based enzyme histochemistry of ChAT could be a useful tool for studying cholinergic neurons.

Original languageEnglish
Pages (from-to)2079-2087
Number of pages9
JournalACS Chemical Neuroscience
Volume12
Issue number12
DOIs
Publication statusPublished - 2021 Jun 16

Keywords

  • choline acetyltransferase
  • cholinergic neurons
  • enzyme histochemistry
  • Mass spectrometry imaging
  • spinal cord injury

ASJC Scopus subject areas

  • Biochemistry
  • Physiology
  • Cognitive Neuroscience
  • Cell Biology

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