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Immunocytochemical Study of Mammalian Progesterone Receptor Using Monoclonal Antibodies^*

Groupe de Recherches sur la Biochimie Endocrinienne et la Reproduction INSERM U.135, Faculté de Médecine Paris-Sud, 94270 Le Kremlin-Bicetre, France

Address requests for reprints to: Dr. M. Perrot-Applanat, INSERM U. 135, Universite Paris-Sud, Faculte de Medecine de Bicetre 78, rue du General Leclerc, Le Kremlin-Bicetre 94270 France.

Abstract

Five monoclonal antibodies were used for the immunocytochemical study of mammalian progesterone receptor (PR). Initial studies were aimed at defining the optimal experimental conditions for the detection of the receptor, with special emphasis on techniques likely to be used in clinical determinations and in immunoelectron microscopic localization. Specific immunoperoxidase staining was observed either in fixed, frozen sections or in sections of paraffin-embedded tissue. The latter method allowed a better preservation of cellular structures. Among the eight fixatives tested, glutaraldehyde, picric-acid formaldehyde, and paraformaldehyde proved satisfactory. Both indirect immunoperoxidase and the indirect antibody peroxidase-antiperoxidase methods could be used.

In immature rabbits or castrated guinea pigs primed by estrogen, i.e. in conditions where its ligand was absent (or present in very low concentration), the PR was confined to the nucleus of immunoreactive cells. This was the case for all the cell types of the endometrium and the myometrium, for the immunostained cells of the oviduct, cervix, vagina, pituitary gland, and for the very weakly stained cells in the liver. No staining was observed in nontarget tissues for progesterone, such as diaphragm, spleen, and small intestine. Nuclear staining was also absent when various control antibodies replaced anti-PR antibodies. This result thus generalizes the observations made on the estrogen receptor, showing that there is no translocation of the receptor from cytoplasm to nucleus under the influence of the hormone. Moreover, a marked heterogeneity in immunostaining was observed among cells of the same type in several tissues, suggesting that there could be large differences in the hormonal sensitivity of individual cells.

Cellular distribution of PR immunoreactivity was studied in the uterus, cervix, oviduct, and pituitary gland of rabbits and in the uterus and vagina of guinea pigs. A labeling was observed in all the cell types of the uterus (luminal and glandular epithelium, stroma, and muscularis). In the cervix, nuclear immunostaining was observed in the connective tissue of the lamina propria and in some epithelial and muscle cells. In the vagina, PR immunoreactivity was seen in the basal layers of the stratified squamous epithelium, in the connective tissue of the lamina propria, and in the smooth muscle. In the oviduct, the luminal epithelium, the connective tissue, and the muscularis were stained. In the pituitary gland, selective nuclear labeling was observed in a few scattered cells. (Endocrinology 116: 1473–1484, 1985)

Footnotes

^* This work was supported by the Institut National de la Sante et de la Recherche Medicale (PRC 135050), the Centre National de la Recherche Scientifique, the Fondation pour la Recherche Médicale, and the Unité d' Enseignement et de Recherches Kremlin-Bicetre.

Received July 24, 1984.

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