Immunocytochemical localization of the heparin-binding growth-associated molecule (HB-GAM) in the developing and adult rat cerebellar cortex

Abstract

The heparin-binding, growth-associated molecule (HB-GAM) is a developmentally regulated protein that belongs to a new family of heparin-binding molecules, not related to the fibroblast growth factors (FGFs), with putative functions during cell growth and differentiation. In order to further study the functional role of HB-GAM we have used a polyclonal antiserum, raised against the purified protein to localize HB-GAM in the developing and adult rat cerebellar cortex. During postnatal development HB-GAM-like immunoreactivity (IR) was found to be present in all layers of the cerebellar cortex. IR was mainly associated with processes or extracellular structures but not with cell bodies. Throughout all the stages examined the molecular layer was clearly labeled, whereas staining in the internal granular layer was diffuse. IR in the external granular layer on postnatal day 1 and 8 was found to be associated with radially oriented fibres connecting the internal granular layer with the pial surface of the cerebellum. The intensity of this staining seemed to increase from day 1 to 8. Staining of corresponding areas with an antiserum against the glial fibrillary acidic protein (GFAP) suggested that the HB-GAM antiserum in the developing cerebellar cortex labers Bergmann glia fibres of Golgi epithelial cells. Because of the diffuse staining of the molecular layer in the adult rat it was not possible to distinguish whether radial fibres in the adult contained any HB-GAM IR. Golgi epithelial cells are considered as crucial for the migration of granular cells during the differentiation of the cerebellar cortex. We therefore speculate that the association of HB-GAM-like IR may be of functional relevance. The fact that molecules, such as tenascin, known to be involved in morphogenetic events show a similar spatial temporal distribution pattern further underscores this hypothesis. HB-GAM, which possesses a classical signal sequence, might be released in the extracellular space and could mediate adhesion phenomena by binding to heparin-like molecules associated with the neuronal membrane. Therefore, it will be important to investigate whether specific antibodies against HB-GAM are able to interfere with normal cerebellar development in vitro and in vivo.