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Journal of Neuroscience, Vol 15, 1329-1340, Copyright © 1995 by Society for Neuroscience
The influence of heregulins on human Schwann cell proliferation
AD Levi, RP Bunge, JA Lofgren, L Meima, F Hefti, K Nikolics and MX Sliwkowski
Miami Project to Cure Paralysis, University of Miami School of Medicine, Florida 33136.
The use of Schwann cell (SC) autotransplantation to influence neural repair
in humans is dependent upon identifying mitogens that will effectively
expand human Schwann cells (SCs) in culture. The recent purification and
molecular cloning of glial growth factor (GGF), a potent mitogen for rat
Schwann cells, has led to the recognition that a family of proteins
(GGF/HRG/NDF/ARIA) are alternatively spliced products of a single gene. The
heregulins (HRGs) have been characterized with respect to their influence
on human breast cancer cell lines; here we examined whether the HRGs have
mitogenic activity for human SCs. Using DNA synthesis assays and serial
passaging of cells in culture, we demonstrate that HRG is an effective
mitogen for human SCs and that, in the presence of agents that elevate
cAMP, it is possible to expand these cells over multiple passages without
overwhelming fibroblast contamination. One putative target for this family
of proteins is p185erbB2, and EGF-like receptor tyrosine kinase that is
encoded by the erbB2 protooncogene. In this report we also demonstrate that
the erbB2/3/4 messages as well as the erbB2/3 receptor proteins are present
within cultured human SCs. The addition of HRG to human SCs results in
tyrosine phosphorylation of a 185 kDa protein. In the presence of
stimulatory concentrations of HRG, a blocking monoclonal antibody (2C4) to
p185erbB2 is capable of significantly inhibiting phosphorylation of a 185
kDa protein as well as the subsequent incorporation of 3H-thymidine within
the human SC. These latter results implicate an important role for
p185erbB2 in mediating the mitogenic response of human SCs to HRGs.
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