A monoclonal antibody, Py, distinguishes different classes of hippocampal neurons

A monoclonal antibody, Py, was produced by immunizing mice with a glycoprotein fraction isolated from 3-week-old rat hippocampus. Py antibodies gave strong immunocytochemical staining of the perikarya and dendrites of large neurons in many areas of the rat brain, including the cerebral cortex, hippocampus, cerebellum, brain stem, and olfactory bulb. Immunoelectron microscopy showed the antigen to be predominantly intracellular, although its presence on the neuronal cell surface was not excluded. The antibody gave differential staining of adult hippocampal neurons, large pyramids of field CA3 being strongly immunoreactive, while CA1 pyramids and the dentate granule cells were unstained. Some interneurons were positive in each of the hippocampal fields. In developing hippocampus, the Py antigen appeared by the middle of the first postnatal week, and the adult pattern of staining was achieved by the end of the second week. Immunoblotting showed the antigen to have a relative mobility of 146 kDa with an additional faint band at 166 kDa. Differential Py staining of neurons was seen in dissociated cultures of embryonic hippocampus and in subdissected hippocampal fragments transplanted into adult host brains. This antibody can therefore be used for identification of hippocampal neurons that have been removed from their normal anatomical context.