Journal of Neuroscience, Vol 9, 2170-2181, Copyright © 1989 by Society for Neuroscience
A monoclonal antibody, Py, distinguishes different classes of hippocampal neurons
PL Woodhams, M Webb, DJ Atkinson and PJ Seeley
Norman and Sadie Lee Research Centre, National Institute for Medical Research, London, England.
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.