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Journal of Histochemistry and Cytochemistry, Vol. 50, 641-650, May 2002, Copyright © 2002, The Histochemical Society, Inc.

ARTICLE

Differences in Postmortem Stability of Sex Steroid Receptor Immunoreactivity in Rat Brain

Mariann Fodora,b, Fred W. van Leeuwena, and Dick F. Swaaba
a Netherlands Institute for Brain Research & Graduate School Neurosciences Amsterdam, Amsterdam, The Netherlands
b Research Institute for Endocrinology, Reproduction and Metabolism, Amsterdam, The Netherlands

Correspondence to: Mariann Fodor, Dept. of Pediatrics, Vrije Universiteit Medical Center, 1007 MB Amsterdam, The Netherlands. E-mail: fodor@vumc.nl

Difficulties in demonstrating sex steroid receptors in the human brain by immunohistochemistry (IHC) may depend on postmortem delay and a long fixation time. The effect of different postmortem times was therefore studied in rat brain kept in the skull at room temperature for 0, 6, or 24 hr after death. After a long fixation for 20 days, hypothalami were embedded in paraffin and sections were immunohistochemically stained for androgen receptor (AR), estrogen receptor-{alpha} (ER), or progesterone receptor (PR). Retrieving the antigenic sites by microwave pretreatment was essential to obtain successful IHC in all groups studied. In general, immunoreactivity was restricted to the cell nuclei. However, the intensity of the staining appeared to be strongly dependent on the different receptor antigens and postmortem time. Both AR and ER but not PR immunoreactivity were decreased after immersion-fixation compared to the perfused sections at time point zero. In brains fixed by immersion, all three receptors decreased gradually with increasing postmortem time, and ER became hardly detectable after 24 hr postmortem. The results of these experiments show that, with the protocol used, postmortem variables and lengthy fixation do not, in principle, prevent sex steroid receptor IHC in human material. The outcome of the immunostaining, however, might be strongly dependent on the epitopes and/or antibody used. (J Histochem Cytochem 50:641–649, 2002)

Key Words: estrogen receptor-{alpha}, androgen receptor, progesterone receptor, immunohistochemistry, postmortem delay, rat


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