Requirement of the hyaluronan receptor RHAMM in neurite extension and motility as demonstrated in primary neurons and neuronal cell lines

Abstract

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.

Migration of bovine aortic smooth muscle cells after wounding injury. The role of hyaluronan and RHAMM.

“…However, the immunolocalization data provide strong evidence that links the initial 12 h of increased motility following injury with the redistribution of RHAMM to the cell surface followed by its disappearance so that, by 18 h, only nuclear staining was observed. We have previously correlated rate of locomotion with cell surface localization of RHAMM in other cell types (27,43,44). The significance of the nuclear staining is unknown, but has previously been noted in fibroblasts (27).…”

Identification of GRP75 as an Independent Favorable Prognostic Marker of Neuroblastoma by a Proteomics Analysis

“…The GRP75/78-RHAMM complex could then bind to the microtubules to stabilize the microtubules in the interphase and prevent the depolymerization of microtubules for the progression of mitosis (32). The essential role of RHAMM in neurite extension has been suggested (33), and the expression of RHAMM has been linked to the progression and metastasis of a variety of cancers (34). Our preliminary data show that SH-SY5Y cells treated with ATRA do exhibit reduced expression of RHAMM, correlating with enhanced neuronal differentiation (data not shown).…”

An Essential Role for the Interaction Between Hyaluronan and Hyaluronan Binding Proteins During Joint Development

“…The monoclonal antibody IVd4 (a kind gift from Prof. Bryan Toole, Tufts University, Boston, MA) was raised against an HABP isolated from embryonic chick brain (Banerjee and Toole 1991). Antibody R7.2 (a kind gift from Prof. Eva A. Turley, Institute of Cell Biology, Manitoba, Canada) was raised against a 12-peptide sequence of rabbit receptor for HA-mediated migration (RHAMM) (Nagy et al 1995). Antibodies CR22, M11, and M22 (gifts from Dr. S. Tsukita) were raised against baby hamster kidney cell ERM proteins (Tsukita et al 1989;Sato et al 1991).…”