NERVE GROWTH-FACTOR (NGF) CONTENT IN ADULT-RAT BRAIN-TISSUES IS SEVERAL-FOLD HIGHER THAN GENERALLY REPORTED AND IS LARGELY ASSOCIATED WITH SEDIMENTABLE FRACTIONS

Initial studies had revealed that the bioactivity of nerve growth fact or (NGF) in sonicates of adult rat hippocampal formation (HF) is sever al-fold greater in their pellet than their supernatant fractions. Such observations have prompted an analysis of NGF antigen (NGF-Ag) conten ts in pellets and supernatants from a variety of adult rat CNS tissues , both in the absence and the presence of exogenous beta-NGF. With HF tissues, NGF-Ag in the supernatants was comparable to most literature values, but pellet NGF-Ag was 3 to 5 times that amount. All other CNS tissue sonicates also revealed 3-6 fold higher NGF-Ag in their pellets than their supernatants, hence overall NGF-Ag contents were greatly i n excess of reported ones. Presentation of mouse beta-NGF to a tissue, its sonicate, or its standard pellet resulted in a transfer to the fi nal pellet of 30-50% of the added soluble NGF-Ag (and 30% of the added bioactivity). This percentage is much lower than that present in nati ve pellets (80%), suggesting that the association of endogenous NGF wi th particulate matter may involve at least two compartments. Treatment s of pellets with salt, alkaline pH, and/or the detergent Triton X-100 have revealed a third subset, namely additional pellet NGF-Ag that wa s not initially recognized by the antibody in our ELISA assay. The tre atments also caused substantial release of NGF from pellet to supernat ant. Further studies should clarify the nature of the association betw een NGF and the three subsets of pellet NGF and allow the investigatio n of the pellet molecules responsible for it.