Identification of sequence, protein isoforms, and distribution of the hyaluronan-binding protein RHAMM in adult and...

The protein RHAMM (for "receptor for hyaluronan-mediated motility"; CD168) is a member of the hyaladherin family of hyaluronan-binding proteins. RHAMM has a role in cell signaling, migration, and adhesion via interactions with hyaluronan, microtubules, actin, calmodulin, and components of the extracellular regulated kinase (erk) signaling pathway. Based on previous findings of potentially similar roles in neural cells in culture, we investigated the molecular characteristics, protein expression profile, and distribution of RHAMM in rat brain. Reverse transcriptase-polymerase chain reaction (RT-PCR) using RNA isolated from adult rat brain yielded a single RHAMM sequence of 2.1 kilobases encoding a protein of 82.4 kDa. RHAMM is subject to alternate splicing in other systems, but no RT-PCR evidence was found for splice variants in brain, although our analysis does not rule out this possibility. The amino acid sequence displayed homology with human and murine RHAMM (74% and 80%, respectively) but contained only one copy of a 21-amino-acid sequence that is repeated five times in the murine homologue. By using anti-RHAMM antibodies, several RHAMM isoforms were identified in brain. Immunohistochemically, RHAMM was found in the vast majority of neurons and in many oligodendrocytes throughout brain, with heterogeneous levels among cell populations, and was confined to the somata and initial processes of these cells. RHAMM was detected in neurons of cerebral cortex and most subcortical and brainstem structures at postnatal day 1 and exhibited an adult distribution pattern by postnatal day 5. High levels were detected in oligodendrocytes by postnatal day 10. The widespread expression of RHAMM in adult and developing brain implies a role for this protein and its ligand hyaluronan in key events of cell signaling and cytoskeletal regulation in the CNS.