Central boutons of glomeruli in the spinal cord of the cat are enriched with L-glutamate-like immunoreactivity.
Several lines of evidence indicate that L-glutamate may be a neurotransmitter of fine myelinated and unmyelinated primary afferent fibres in the spinal cord. The aim of the present study was to determine if L-glutamate was enriched in the terminals of these fibres. We performed the post-embedding immunogold technique on sections taken from the superficial regions of the lumbar cord in two cats. An antiserum, raised against protein-conjugated L-glutamate, was employed. Several tests on tissue and on a model system indicated that the antiserum recognized a glutaraldehyde-fixed L-glutamate-like substance. Terminals of fine afferent fibres were identified in the substantia gelatinosa as central boutons of synaptic glomeruli. Central boutons were examined through serial sections following immunogold reactions and were found to be heavily labelled with gold particles in consecutive sections. Quantitative analysis indicated that central boutons were more than two and a half times as densely labelled with gold particles than the tissue average. It was concluded that this represents a genuine enrichment of L-glutamate in these structures. Comparisons were made between L-glutamate-immunoreactive properties of central terminals and immunoreactivity for GABA, aspartate and glutamine. Statistical analysis revealed that central boutons in sections incubated in GABA antiserum and glutamine antiserum were associated with significantly lower densities of gold particle labelling than the average for the same tissue. Particle densities of central boutons in sections incubated in aspartate antiserum were not significantly different from average tissue densities. It was concluded that central boutons were not enriched with these three amino acids. Central boutons of synaptic glomeruli were classified into three groups on morphological criteria: (1) dense sinusoidal boutons; (2) large dense-core vesicle-containing boutons; and (3) regular synaptic vesicle-containing boutons. Quantitative analysis revealed that all of these groups were enriched in glutamate immunoreactivity, however, there were differences between the groups; large dense-core vesicle-containing boutons were associated with significantly lower densities of particles than regular synaptic vesicle-containing and dense sinusoidal terminals. The evidence indicates that central boutons, which most probably originate from fine myelinated and unmyelinated primary afferent fibres, are enriched with L-glutamate which may serve as a neurotransmitter in such fibres.
Department of Preclinical Veterinary Sciences, University of Edinburgh, U.K.
D J
DJ
Maxwell
W M
WM
Christie
A D
AD
Short
J
J
Storm-Mathisen
O P
OP
Ottersen
D000818:Animals; D000918:Antibody Specificity; D002415:Cats; D005971:Glutamates / Q000276:immunology / Q000378:metabolism*; D006046:Gold; D007150:Immunohistochemistry; D009411:Nerve Endings / Q000166:cytology / Q000276:immunology; D013116:Spinal Cord / Q000166:cytology / Q000378:metabolism*; D013194:Staining and Labeling
D005971:Glutamates; D006046:Gold