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Journal of Neuroscience, Vol 8, 472-492, Copyright © 1988 by Society for Neuroscience
Glycine and glycine receptor immunoreactivity in brain and spinal cord
AN van den Pol and T Gorcs
Section of Neurosurgery, Yale University School of Medicine, New Haven, Connecticut 06510.
To study the distribution of glycine immunoreactive neurons in the spinal
cord and brain, antisera were raised against glycine conjugated to protein
carriers. High-titer rabbit glycine antiserum was purified by affinity
chromatography. Testing against other amino acids and peptides with immuno
dot blots and ELISA assays showed little apparent cross-reaction with
glutamate, aspartate, glutamine, taurine, and 17 other amino acids and
related compounds. Similarly, the antiserum showed little apparent
recognition of glycine when glycine was incorporated into peptides. A
slight cross-reactivity with GABA, beta- alanine, and cysteine was found.
Immunocytochemical labeling of tissue sections could be blocked with
glycine conjugated to a heterologous carrier protein but not by other amino
acids conjugated to that protein. Immunocytochemistry at the light
microscope level with immunofluorescence and silver-intensified colloidal
gold revealed a wide distribution of glycine-like immunoreactivity
throughout all laminae of the rat spinal cord and in all segments studied
from the cervical, thoracic, lumbar, and sacral cord. Immunoreactive
boutons were found terminating on both cell bodies and on dendrites.
Ultrastructural analysis with postembedding colloidal gold
immunocytochemistry demonstrated large numbers of immunoreactive boutons
making symmetrical type synapses with neuronal perikarya, including motor
neurons, and with proximal and distal dendrites. Presynaptic glycine
immunoreactive boutons were found in both ventral and dorsal horn.
Immunoreactivity was concentrated over regions rich in vesicles, and over
mitochondria in immunoreactive boutons, but not over mitochondria in
postsynaptic dendrites. Glycine-immunoreactive perikarya were identified
both in the dorsal horn and in the ventral horn. Myelinated and
unmyelinated glycine-immunoreactive axons were noted both in the gray and
white matter of the cord. The density of immunoreactive axons varied in the
white matter, with the greatest number of immunoreactive axons found in the
white matter adjacent to the gray matter in lateral and ventral white.
Significantly fewer immunoreactive axons were found in the white matter of
the dorsal columns. Myelin sheaths around axons were unlabeled. The
distribution of glycine-immunoreactive boutons correlated well with the
distribution of glycine receptor immunoreactivity on postsynaptic elements
of the spinal cord, tested with different monoclonal antisera against
strychnine-purified glycine receptor. Glycine receptor immunoreactivity was
found throughout the gray matter of both rat and primate.(ABSTRACT
TRUNCATED AT 400 WORDS)
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