Nuclear location of hormone-free estrogen receptors by monoclonal antibodies could be a tissue-fixation dependent artifact.
The 'two-step' model proposed by Jensen and his collaborators for explaining estrogen action conceptualized hormone-free estrogen receptors (ER) to be cytoplasmic, and hormone-filled, transformed ER to be nuclear. Applying monoclonal antibodies which recognized epitopes in ER and formaldehyde-fixed tissues, King et al demonstrated exclusively nuclear staining in target tissues utilizing immunoperoxidase technique. Recently these antibodies have become commercially available enabling other investigators to conduct studies. In this report, using these monoclonal antibodies we have demonstrated that a change in the concentration of formaldehyde alters the staining pattern yielding cytoplasmic instead of nuclear staining in calf uterus, MCF-7 cells, and ER(+) human breast cancer. In addition, neutralization of the antibody activity was not achieved with freshly prepared ER(+) cytosols. Formaldehyde-treated cytosols were essential. These results ought to caution investigators in determining in vivo location of antigens based on the staining pattern obtained in fixed tissues. Furthermore, this effect of formaldehyde on estrogen receptors may be applicable to other steroid hormone receptors.
Raam
S
S
Department of Medicine, Tufts University School of Medicine, Boston, MA.
Lauretano
A M
AM
Vrabel
D M
DM
Pappas
C A
CA
Tamura
H
H
D000818:Animals; D000911:Antibodies, Monoclonal; D002417:Cattle; D002467:Cell Nucleus / Q000032:analysis*; D005404:Fixatives; D005557:Formaldehyde; D006801:Humans; D007150:Immunohistochemistry; D011960:Receptors, Estrogen / Q000032:analysis*; D015203:Reproducibility of Results
D000911:Antibodies, Monoclonal; D005404:Fixatives; D011960:Receptors, Estrogen; D005557:Formaldehyde