A protein encoded by an alternative splice variant of choline acetyltransferase mRNA is localized preferentially in peripheral nerve cells and fibers

J Chem Neuroanat. 2000 Jan;17(4):217-26. doi: 10.1016/s0891-0618(99)00043-5.

Abstract

Central cholinergic systems have been visualized by immunohistochemistry using antibodies to choline acetyltransferase (ChAT). Peripheral cholinergic cells and fibers, however, have been hardly detectable with most of these antibodies. This phenomenon suggests that a different form of ChAT may exist in peripheral tissues. Here we report two types of mRNA for ChAT expressed by alternative splicing in rat pterygopalatine ganglion. One is exactly identical with ChAT mRNA reported in the central nervous system (ChAT of a common type; cChAT). The other lacks exons 6, 7, 8 and 9, which was detected only in the pterygopalatine ganglion (ChAT of a peripheral type; pChAT). The peculiarity of pChAT in chemical structure, possessing a splice joint of the exons 5 and 10, led us to produce rabbit antisera against a recombinant peptide of 41 amino acids which spans over the splice joint. On Western blots using a successfully obtained antiserum, an intense band of about 50 kDa, corresponding to the expected molecular weight of pChAT, was detected in the pterygopalatine ganglion but not in the brain. Immunohistochemistry using the antiserum failed to reveal positive staining of known brain cholinergic structures, while it permitted us to observe peripheral, probably cholinergic, nerve cells and fibers including those in the pterygopalatine ganglion and enteric nervous system.

MeSH terms

  • Alternative Splicing*
  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Choline O-Acetyltransferase / analysis
  • Choline O-Acetyltransferase / chemistry
  • Choline O-Acetyltransferase / genetics*
  • DNA Primers
  • Dihydrolipoamide Dehydrogenase / analysis
  • Exons
  • Ganglia, Parasympathetic / enzymology*
  • Genetic Variation*
  • Immune Sera
  • Immunohistochemistry
  • Male
  • Molecular Sequence Data
  • Nerve Fibers / enzymology*
  • Neurons / enzymology*
  • Peptide Fragments / chemistry
  • Polymerase Chain Reaction
  • RNA, Messenger / genetics*
  • Rabbits
  • Rats
  • Rats, Wistar

Substances

  • DNA Primers
  • Immune Sera
  • Peptide Fragments
  • RNA, Messenger
  • Dihydrolipoamide Dehydrogenase
  • Choline O-Acetyltransferase