43] Simultaneous localization of steroid hormones and neuropeptides in the brain by combined autoradiography and immunocytochemistry

Abstract

Publisher Summary This chapter describes the combined technique of steroid hormone autoradiography and immunocytochemistry. In the combined technique of steroid hormone autoradiography and immunocytochemistry, autoradiograms are prepared by the thaw-mount or dry-mount technique after injection of radioactively labeled hormone. Subsequently the autoradiograms are stained by the peroxidase-antiperoxidase (PAP) immunocytochemical staining technique using unlabeled antibodies. Some of the steps for the preparation of autoradiograms are identical in both thaw-mount and dry-mount techniques, including freezing, storage, and sectioning of tissues as well as photographic development and fixation. The technique of autoradiography and immunocytochemistry for localization of radioactively labeled steroid hormones and antibodies to protein hormones, neuropeptides, or neurotransmitter synthesizing enzymes in the brain. This method enables the simultaneous visualization of radioactively labeled cells and peptide or neurohormone producing cells in the same tissue preparation. With this procedure, anatomical relationships between sites of action and sites of production of two different substances have been demonstrated.