Hyaluronan was studied in extracts of the nervous system and in situ. Extraction yielded larger amounts at neutral or alkaline pH. Protease digestion enhanced the quantitative results obtained with an indirect enzyme immunological assay. It was shown that HA extracted from brain at neutral pH was bound to a glycoprotein component (hyaluronectin, HN) which is in part free at acid pH. Although HN is not restricted to nervous tissue it is mainly expressed in the central nervous system of adult mammals. Its main form has a molecular mass of 68 kDa and binds in vitro to HA and to HA-derived oligosaccharides down to HA10 with a Kd in the 10(-8) M range. HA-HN complexes were found in human cerebrospinal fluids. The HA concentration in cerebral tissue decreases from the fetus to the adult, whereas the HN concentration increases. HA is not however saturated by HN and still binds HN in vitro. In the rat HA decreases sharply at Days 10-11 after birth. In the rat embryo HA forms an extracellular component of the migration and proliferation areas of the cerebral cortex. In the adult typical locations were at the nodes of Ranvier and in perineuronal structures. HN was found in the same locations but seemed to be associated with a restricted category of neurons. In the cerebellum HA-HN was found mainly in the grey nuclei, the granular layer and around Purkinje cells. Cell bodies were not stained but in the electron microscope HN was seen in the cytoplasm and plasma membrane of the perisynaptic glial cell processes. A hypothesis has been proposed that HA-HN is involved in neural GABAergic transmission.