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Journal of Neuroscience, Vol 14, 3713-3724, Copyright © 1994 by Society for Neuroscience
Neuronal expression of glypican, a cell-surface glycosylphosphatidylinositol-anchored heparan sulfate proteoglycan, in the adult rat nervous system
ED Litwack, CS Stipp, A Kumbasar and AD Lander
Department of Biology, Massachusetts Insitute of Technology, Cambridge 02139.
Cell-surface proteoglycans have been implicated in cell responses to growth
factors, extracellular matrix, and cell adhesion molecules. M12, one of the
most abundant membrane-associated proteoglycans in the adult rat brain, is
a approximately 65 kDa glycosylphosphatidylinositol- linked protein that
bears heparan sulfate chains (Herndon and Lander, 1990). To assess its
identity, M12 was purified and internal peptide sequences obtained.
Comparison of the results with protein sequence predicted by a cDNA cloned
from PC12 cells indicated that M12 is rat glypican, a proteoglycan first
cloned from human fibroblasts. In addition, antibodies raised against a rat
glypican fusion protein specifically detected the 65 kDa brain proteoglycan
core protein, both by immunoprecipitation and by Western blotting. Northern
blot analysis using a rat glypican probe also detected glypican message in
the adult, as well as the developing rat brain. In situ hybridization with
glypican RNA probes showed that glypican is expressed in a subset of
structures in the adult rat nervous system. These include the hippocampus,
dorsal thalamus, amygdala, cerebral cortex, piriform cortex, olfactory
tubercle, several cranial nerve nuclei, the ventral horn of the spinal
cord, and the dorsal root ganglia. Several other brain regions exhibited
little or no hybridization over background. In most cases where glypican
hybridization was observed, the signal could be localized specifically to
the cell bodies of identifiable neurons, for example, spinal motoneurons,
hippocampal pyramidal cells. In the cerebral cortex, glypican hybridization
was found in layers 2/3, 5, and 6, but was missing from 1 and 4. The data
suggest that glypican is expressed primarily by subpopulations of
projection neurons in the adult rat nervous system.
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