Laurel E. Glass

Localization of autologous and heterologous serum antigens in the mouse ovary.

Abstract The ovarian localization of systemically injected bovine plasma albumin, rabbit globulin, and autologous serum antigens was examined in the mouse by the fluorescent antibody technique. Bovine plasma albumin injected intravenously was localized primarily in the follicular fluid, granulosa cell cytoplasm, zona pellucida, and oocyte in tertiary follicles and in the ovarian connective tissues. It was not detected in primary and secondary follicles nor in corpora lutea. Rabbit globulin was not detected in the ovary except when injected intraperitoneally at relatively high concentrations; localization in such instance was in low amount in the follicular fluid and granulosa cell cytoplasm of tertiary follicles. Antigenic molecules similar to or identical with mouse serum macromolecules were detected in the cytoplasm of most ovarian cell types including the oocytes of primary, secondary, and tertiary follicles. In addition, the autologous serum molecules were present in the follicular fluid and zona pellucida of tertiary follicles; they were not detected in ovarian connective tissues. These data demonstrate that antigens may be transferred from maternal serum to the oocyte in the mouse; they suggest strongly that macromolecular transfer occurs. It is hypothesized that similar serum-to-oocyte transfer of large molecules synthesized by the maternal organism is a necessary concomitant of normal mammalian oogenesis.

Studies of induced malignant tumors of placental and uterine origin in the rat

Abstract 1. Induced malignant tumors of placental and uterine origin were studied in the rat. Dimethylbenzanthracene dissolved in beeswax was applied to 1,042 embryonic sacs of 144 adult Long-Evans rats after fetectomy on the tenth to twelfth day of pregnancy. At various intervals (7 to 40 weeks) following the operation, the rats were sacrificed and the induced tumors and their metastases were examined histologically. 2. Malignant tumors were induced in 65 rats (45 per cent). Eleven pulmonary metastases (17 per cent) and 10 lymph node metastases (15.4 per cent) were observed in the tumor-bearing animals. 3. Seven choriocarcinomas were produced, an incidence of less than 1 per cent of all embryonic sacs. Three out of the 7 choriocarcinomas had pulmonary metastases, whereas no lymph node metastasis was found. 4. As to malignant tumors of uterine origin, 7 sarcomas (leiomyosarcomas and endometrial stromal sarcomas), 2 carcinosarcomas, 46 malignant tumors forming glandular spaces (adenocarcinomas and malignant mesotheliomas), 4 adenoacanthomas, and 16 squamous cell carcinomas were induced. The pathogenesis of these induced tumors is discussed.Abstract 1. Induced malignant tumors of placental and uterine origin were studied in the rat. Dimethylbenzanthracene dissolved in beeswax was applied to 1,042 embryonic sacs of 144 adult Long-Evans rats after fetectomy on the tenth to twelfth day of pregnancy. At various intervals (7 to 40 weeks) following the operation, the rats were sacrificed and the induced tumors and their metastases were examined histologically. 2. Malignant tumors were induced in 65 rats (45 per cent). Eleven pulmonary metastases (17 per cent) and 10 lymph node metastases (15.4 per cent) were observed in the tumor-bearing animals. 3. Seven choriocarcinomas were produced, an incidence of less than 1 per cent of all embryonic sacs. Three out of the 7 choriocarcinomas had pulmonary metastases, whereas no lymph node metastasis was found. 4. As to malignant tumors of uterine origin, 7 sarcomas (leiomyosarcomas and endometrial stromal sarcomas), 2 carcinosarcomas, 46 malignant tumors forming glandular spaces (adenocarcinomas and malignant mesotheliomas), 4 adenoacanthomas, and 16 squamous cell carcinomas were induced. The pathogenesis of these induced tumors is discussed.

Studies of induced malignant tumors of placental and uterine origin in the rat: I. Survival of placental tissue following fetectomy

Abstract 1. The survival of placental and embryonic tissues after fetectomy was studied in the rat. At the ninth, tenth, eleventh, or twelfth day of pregnancy, fetuses were removed from gestation sacs in the left uterine horn of 46 adult Long-Evans rats; after fetectomy, a beeswax pellet was inserted and the sac was ligated. At the eighteenth, twenty-fifth, thirty-second, or fortieth day after conception, the rats were sacrificed and the fetectomized gestation sacs were removed, sectioned, and examined histologically. 2. Trophoblastic cells regressed within 3 weeks after fetectomy in most of the embryonic sacs (94.1 per cent) although they survived in the myometrium of 5.4 per cent for more than 4 weeks after fetectomy. Cortisone injection had no effect on trophoblast survival. Yolk sac epithelium survived only 2 weeks after fetectomy in 4.9 per cent. 3. Embryonic cells, injected subcutaneously into the mothers at the time of operation or persistent in the uterine wall, survived to the fortieth day after conception in 36.4 per cent of the animals; fetal tissues had developed in all of these. The survival rate of embryonic cells was much higher than that of trophoblastic cells. 4. No differences in the survival of trophoblast, yolk sac, or embryonic tissue were observed whether animals were fetectomized on the ninth, tenth, eleventh, or twelfth day of pregnancy.

Studies of induced malignant tumors of placental and uterine origin in the rat: III. Identification of experimentally induced choriocarcinoma by detection of placental hormone

Abstract In order to verify a placental derivation, the luteotropic and mammotropic activities of experimentally induced choriocarcinoma were examined by the deciduoma test in traumatized endometrium and by the induction of lobuloalveolar proliferation of the mammary glands of hypophysectomized rats. All 5 assay animals injected with the tissue extract of a choriocarcinoma showed mammotropic activity (lobuloalveolar growth of mammary gland). Nineteen induced malignant tumors other than choriocarcinoma had no such hormonal activities by these tests. These results lead to the convincing conclusion that the induced tumors which were diagnosed as choriocarcinoma histologically are true choriocarcinomas. Our findings also appear to favor the concept that the source of luteomammotropic substance is the trophoblastic giant cell.

ELECTROPHORESIS OF SERUM PROTEINS AND SELECTED ENZYMES IN MALES, NON-PREGNANT, PREGNANT, AND LACTATING FEMALE MICE.

Summary Serum proteins and selected enzymes in non-pregnant, pregnant, lactating females and in male mice were studied by starch gel electrophoresis. Esterase and cholinesterase activity were present in all protein regions; alkaline and acid phosphatase, aminopeptidase, and cytochrome oxidase activity was associated with the protein beta region. Total protein was approximately the same in all categories of sera studied, but zymograms and protein stained gels showed that the relative concentrations of certain serum components were different in different sex, pregnancy or lactation categories. The relationship between these variations and possible transfer of maternal serum molecules to the oocyte or embryo is discussed.

An Immunologic Approach to Contraception: Localization of Antiembryo and Antizona Pellucida Serum during Mouse Preimplantation Development**Supported by National Institute of Child Health and Human Development Contract NIH 70-2309.

Rabbit antisera were developed to mouse preimplantation eggs at 3 developmental stages: 1) to the unfertilized just ovulated cumulusoocyte mass; 2) to morulae; and 3) to uterine blastocyst. Unabsorbed antisera and antisera absorbed with whole mouse serum were characterized against homologous and heterologous embryonic antigen and against whole mouse serum by immunoelectrophoresis by in vitro embryo culture and by fluorescent antibody methods. In fluorescent antibody studies each antiserum localized slightly on ooplasm at homologous stages and strongly on the zona pellucida at all ovarian and preimplantation stages. Embryos died when cultured in media containing antiembryo sera. The possiblity that antizona pellucida serum could be developed as a contraceptive antibody in instances of single coitus was discussed. Also it was suggested that the zona pellucida may be usable as a reproduction-specific antigen for direct contraceptive immunization of the female.