Localization of Immunoreactive Female Steroids in Ovarian and Placental Tissue

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Immunohistochemical localization of estrone (E₁) and progesterone (P)in porcine ovary and human placenta has been demonstrated by direct and indirect antibody techniques. E₁-bovine serum albumin (BSA) and P-BSA conjugates have been used for the immunization of rabbits. The antisera obtained were titrated by the passive hemagglutination test (E₁: 1: 655,360, P: 1: 327,680) and radioimmunoassay measurement ability of various dilutions of antisera (E₁: 1: 30,000, P: 1: 40,000). The anti-steroid properties of the sera indicated that they were specific for each S-P conjugate, and“link-antibody”was formed between the antigen and BSA as revealed by the immunodiffusion technique due to the precipitin line observed using the BSA-absorbed antiserum. In the Graafian follicle, both steroids have been demonstrated within round or oval single cells of the theca interna, although the intensity of the reaction was higher with the E₁ immunity, and correlated with the localization of sudanophilia, G-6-PDH,3β-ol SDH, and lysosomal enzymes. The cells appeared to be theca gland cells. A similar reaction was observed in the majority of the interstitial cells including the hilar cells. Some groups of these, however, exhibited distinct granular and orange colored autofluorescence, and showed contours of macrophages that had engulfed steroid producing cells. In the corpora lutea, the same steroids were found in the theca-lutein cells. The granulo-lutein cells had no antigenicity to E₁, but showed a very weak immuno-positive reaction to P, moderate sudanophilia, and strong activities of G-6-PDH and 3β-ol SDH. The atretic follicles and interstitial glands were immuno-positive to E₁. In human placenta at term, E₁and P were localized at the syncytiotrophoblast layer, although the Langhans cell region was not identified. However, a distinct and unequivocal identification between the two steroids investigated was not possible.