Light and electron microscopic immunohistochemical localization of N- acetylaspartylglutamate (NAAG) in the olivocerebellar pathway of the rat

Waleed M. Renno, Jang Hern Lee, Alvin J. Beitz

Research output: Contribution to journalArticle

29 Scopus citations

Abstract

The inferior olive (IO) is the sole contributor of climbing fibers (CF) to the Purkinje cells of the cerebellar cortex. Although the anatomy and the connectivity between the IO and the cerebellum have been well established, there is still controversy regarding the neurotransmitter systems mediating olivocerebellar projections. The excitatory amino acids, glutamate (Glu) and aspartate (Asp), have both been considered as neurotransmitter candidates of olivocerebellar projections in the rat. More recently N- acetylaspartylglutamate (NAAG) has also been proposed as a transmitter of cerebellar climbing fibers based on biochemical and electrophysiological data. The aim of the present study was to determine whether NAAG immunoreactivity is present in the IO and CF at the light and electron microscopic levels and to quantitate the amount of immunogold labeling in olivary neurons and climbing fiber terminals containing this dipeptide. A polyclonal antisera against NAAG was utilized with a peroxidase-labeled avidin-biotin procedure to demonstrate these immunoreactive neurons in the IO at the light microscopic level. Approximately 45% of olivary neurons display NAAG-like immunoreactivity, and their distribution is unevenly clustered throughout the inferior olive. Using postembedding immunogold electron microscopy in combination with quantitative procedures, we found the highest densities of gold particles in the axonal terminals synapsing on olivary neurons (101.0 particles/μm2), in CF terminals (96.3 particles/μm2), and in some mossy fiber terminals (101.0 particles/μm2). Approximately half of the climbing fiber terminals examined were unlabeled. Moderate labeling occurred in CF axons (70.8 particles/μm2), while IO neuronal perikarya were lightly but significantly labeled (41.6 particles/μm2). The localization of NAAG in the subset of cerebellar climbing fiber terminals provides anatomical support for the hypothesis that NAAG may serve as a neurotransmitter/neuromodulator candidate in the olivocerebellar pathway.

Original languageEnglish (US)
Pages (from-to)140-154
Number of pages15
JournalSynapse
Volume26
Issue number2
DOIs
StatePublished - Jun 1997

Keywords

  • NAAG
  • climbing fibers
  • excitatory amino acids
  • immunohistochemistry
  • inferior olive
  • olivocerebellar

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