Two glycoproteins, tentatively designated as PZ-alpha and PZ-beta, have been isolated and purified to homogeneity from porcine zonae pellucidae by a simple purification procedure producing a high yield. The procedure included the dissolution of the zona material in 0.1 M sodium borate buffer pH 10.0, Sephadex G-100 column chromatography and preparative SDS-polyacrylamide gel electrophoresis. The purified glycoproteins gave a single band on polyacrylamide gel and had molecular weights of 60 000 (PZ-alpha) and 96 000 (PZ-beta). Glutamic acid was detected as the NH2-terminal residue in both glycoproteins, using the dansyl chloride method. Though their amino acid compositions were similar, their carbohydrate contents were slightly different (PZ-alpha: 24.9%; PZ-beta; 19.6%), but these components contained the same types of monosaccharides: fucose, mannose, galactose, NAcGlc and sialic acid. The antigenic properties of the two glycoproteins were indistinguishable by immunodiffusion tests. The PZ-beta could be converted in part to smaller molecular weight components, though not to PZ-alpha, by treatment with beta-mercaptoethanol. Thus clear differences between PZ-alpha and PZ-beta could not be detected by chemical or immunological analyses except for the difference in the behaviour on SDS-polyacrylamide gel electrophoresis.

Amino Acids↗

PubMed1983

Interrelation between luteal cell types in steroidogenesis in vitro of human corpus luteum.

Slices of human corpora lutea of menstruation at different stages, or of inner and outer layers consisting of pure granulosa lutein cells and combined theca and granulosa lutein cells respectively, were incubated with [4-14C]-pregnenolone or with [4-14C]-androstenedione respectively. Conversion of 14C-radioactivity to progestins, androgens and/or estrogens was assessed by the reverse dilution technique with recrystallization to constant specific activity. Although progesterone was the major steroid formed from pregnenolone, a transient increase in androgen formation with a concomitant decrease in estrogen formation in developing corpora lutea was followed by an increase in estrogen with a decrease in androgen formation in mature corpora lutea. The reciprocal relation between estrogen and androgen formation is associated with the behaviour of theca lutein cells since the cells disappeared at the former and clearly differentiated at the latter stages of corpora lutea. The outer layers consistently produced more 17-hydroxyprogesterone, androgens and estrogens from pregnenolone than the inner layers, while the inner layers converted constantly more androstenedione to estrogens than the outer layers. It is suggested that aromatase activity of granulosa lutein cells in co-existence with theca lutein cells is required by human corpora lutea for fulfillment of the function of estrogen synthesis.

Androstenedione↗

PubMed1983

Ultrastructural study of minute uterine leiomyomas.

Minute uterine leiomyomas, less than 3 mm in diameter, were studied by electron microscopy. In five of 15 cases, morphologically different types of smooth muscle cell were identified. In the central region of leiomyomas, most myoma cells were characterized by filaments sporadically located in the cytoplasm and well-developed organelles. These cells were interpreted as immature smooth muscle cells. In the outer layer of nodules, the cells were a more mature form of smooth muscle cell and resembled normal myometrial cells. The differences suggest that differentiation of the smooth muscle cells occurs early in the growth of uterine leiomyoma.

Adult↗

PubMed1983

Plasminogen activator in rat ovary during the ovulatory process: independence of prostaglandin mediation.

To elucidate the possible roles of increased plasminogen activator (PA) in follicular rupture and to investigate whether prostaglandins participate in ovarian PA synthesis in vivo, enzyme activities were sequentially measured by a method using the chromogenic substrate S-2251 in immature rat ovaries primed with pregnant mare serum gonadotrophin (PMSG) followed by human chorionic gonadotrophin (hCG) either alone or with a concurrent injection of indomethacin. Before hCG injection PA activity was 0.006 +/- 0.006 (S.D.) mumol/1.6 mg ovarian tissue (wet wt) per 30 min: PA activity of a saline-treated group remained at low levels (less than 0.018 +/- 0.003 mumol/l X 6 mg tissue per 30 min). In contrast, PA activity of animals given hCG alone increased after the treatment, reaching a peak value of 0.112 +/- 0.071 mumol/l X 6 mg tissue per 30 min 12h later, before decreasing to 0.023 +/- 0.014 mumol/l X 6 mg tissue per 30 min at 32 h. Contrary to expectations, a dose of indomethacin which completely blocked ovulation had no effect on either the magnitude or the time-course of PA synthesis after hCG administration (P greater than 0.05). These results indicate that prostaglandins are not involved in the preovulatory synthesis of PA induced by hCG in rat ovaries and that PA is not a primary proteolytic enzyme for follicular rupture. It is suggested that PA has possible roles in cumulus detachment and/or proliferation of granulosa cells during the ovulatory process.

Animals↗

PubMed1983

Discordant pregnancy tests in detection of alpha-subunit--producing tumors.

Ectopic production of isolated alpha-subunit of glycoprotein hormones was detected in a 32-year-old Japanese woman with metastatic lung tumor by a discordance in the immunologic pregnancy tests for native human chorionic gonadotropin (hCG) assays. Quantitative discrepancy of titrations among different types of pregnancy tests, ie, competitive (hemagglutination or latexagglutination inhibition) and noncompetitive (hemagglutination or latexagglutination) reactions, was also confirmed experimentally using a highly purified ectopic alpha-subunit from the patient's urine. Thus the discrepancy between the 2 types of assay systems was due, at least partly, to a cross-reaction of high concentrations (20,000 ng/ml) of ectopically secreted free alpha-subunit of glycoprotein hormones.

Adult↗

PubMed1983

[Tumor markers in gynecologic malignancies].

Human chorionic gonadotropin (hCG) is widely used as a tumor specific index substance for the trophoblastic disease. The radioimmunoassay (RIA) of the C-terminal peptide of hCG-beta subunit can detect hCG up to the level of 0.5 IU/day in urine or 0.5mIU/ml in serum without cross-reactivity with luteinizing hormone. Therefore, this RIA would be used as a hCG-specific assay method in order to ensure the disease is in complete remission and to detect early recurrence. Alpha-fetoprotein and lactic dehydrogenase can be used as a sensitive tumor marker for embryonal carcinoma and dysgerminoma respectively. Carcinoembryonic antigen may serve as a valuable aid in monitoring the response to the therapy of the patient with ovarian or cervical carcinoma, but may not be useful for early detection of these diseases.

Carcinoembryonic Antigen↗