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Journal of Neuroscience, Vol 15, 241-252, Copyright © 1995 by Society for Neuroscience
Requirement of the hyaluronan receptor RHAMM in neurite extension and motility as demonstrated in primary neurons and neuronal cell lines
JI Nagy, J Hacking, UN Frankenstein and EA Turley
Department of Physiology, University of Manitoba, Winnipeg, Canada.
The recently cloned and characterized hyaluronan (HA) receptor RHAMM
(receptor for HA-mediated motility) has been shown to play a critical role
in mechanisms underlying the motile capacity of a variety of peripheral
cell types. Similarities in molecular processes that govern cell locomotion
and growth cone migration prompted us to investigate whether RHAMM also
contributes to neurite migration in vitro. In immunohistochemical studies
of PC12 cells, NG108-15 cells and a neuroblastoma/spinal cord neuronal
hybrid cell line (NSC-34 cells) as well as rat and human primary neurons, a
punctiform RHAMM labeling pattern was detected in cell bodies, along
processes, and at growth cones. By Western blot analysis, the cells lines
expressed major RHAMM forms with apparent MW of 60, 75, and 116 kDa.
Treatment of NG108-15 cells with dibutyryl-cAMP led to a clear increase in
immunolabeling for RHAMM and enhanced expression of the 60 and 75 kDa
forms. A polyclonal anti-RHAMM antibody that interferes with HA/RHAMM
interaction significantly reduced neurite migration of each cell type
examined, while another directed against a RHAMM repeat sequence thought to
promote RHAMM receptor aggregation significantly stimulated neurite
migration of NSC-34 and rat primary neurons. Different monoclonal anti-
RHAMM antibodies had differential inhibitory actions on neurite movement.
Low concentrations (ng/ml) of a peptide corresponding to an HA binding
domain within RHAMM inhibited neurite migration. These results are the
first to implicate RHAMM in the mediation of neurite motility and migration
and to point to the potential importance of HA in this process.
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