Abstract

The expression of monoclonal antibody AB-2 immunoreactivity is age- and sex-dependent in radial glia of developing rat hypothalamus and is regulated by prenatal exposure to gonadal steroids. In the present study, several proteins were recognized by AB-2 and were distributed selectively in subcellular fractions from neonatal hypothalamus (HYP), remaining forebrain (FB), and brainstem regions. Immunoblots revealed polypeptide bands in 3 major molecular weight classes: one at approximately 195 kDa in the cytosolic compartment; and two doublets at 220 kDa and 340 kDa in both microsomal and crude mitochondrial membrane fractions. The 220 kDa and 340 kDa doublets were also Triton-insoluble, suggesting a cytoskeletal association. The 195 kDa-AB-2-immunoreactive band was present in both Triton-soluble and insoluble fractions. AB-2 also recognized several acidic glycolipids extracted from postnatal rat brain regions on immunoblots following high performance thin layer chromatography. One of the bands from postnatal rat brain extracts migrated similarly to purified bovine brain sulfatide, which was also immunoreactive with AB-2. AB-2 immunoreactivity with proteins, polar lipids, and sulfatide suggests that the epitope is a carbohydrate present in multiple cellular compartments. AB-2 recognized the same molecular bands in males and females. Testosterone treatment selectively decreased the level of the 195 kDa AB-2-immunoreactive polypeptide. The 195 kDa AB-2-immunoreactive polypeptide possibly acts in radial glia in the determination of sexually dimorphic neurons in the preoptic area/hypothalamus.