Oviductal Glycoprotein Research Articles

Study of the differentiation of secretory cells in the golden hamster oviductal epithelium by use of a monoclonal antibody

The ontogeny of an oviductal zona pellucida glycoprotein, designated ZP-0, in the golden hamster oviductal epithelium was investigated by means of light and electron microscopic immunolabeling studies using a monoclonal antibody. Light microscopic immunohistochemistry showed that ZP-0 appeared in the oviductal epithelium from 9.5 to 10.5 days after birth. At first, ZP-0 appeared in the Golgi region of the epithelial cells, and then increased in amount as development progressed to fill the supranuclear cytoplasm. Electron microscopic observations showed that rough endoplasmic reticulum and Golgi apparatus were well developed in non-ciliated cells of the ampullar and isthmic segments at 10.5 days after birth. Secretory granules appeared near the Golgi apparatus and markedly increased in number until 15.5 days after birth. Ultrastructural immunocytochemistry showed that ZP-0 appeared in the well-developed Golgi apparatus and then was stored within the secretory granules of developing secretory cells. ZP-0 was never present in the ciliated cells. These results indicate a coincidence between the beginning of ZP-0 biosynthesis and the morphological cytodifferentiation of secretory cells in the golden hamster oviduct during postnatal development. This leads us to the conclusion that ZP-0 is a possible differentiation marker of the golden hamster oviductal secretory cell.

Effect of steroid hormones on sulfated oviductal glycoprotein secretion by oviductal explants in vitro.

Explants of rabbit ampullary and isthmic tissue were cultured 4 days with and without exogenous steroids, and the sulfated oviductal glycoprotein (SOG) concentration in the explant culture supernatants was determined. Tissues cultured with progesterone plus estrogen secreted significantly more SOG than control tissues, whereas tissues cultured with estrogen alone did not. Ampullary tissues cultured with progesterone plus estrogen secreted significantly more SOG than control tissues on Days 2 and 3, whereas SOG secretion by isthmic tissues was significantly above control secretion on Day 4. Ampullary and isthmic tissues differed significantly in their secretory capacity. Maximum ampullary SOG secretion was approximately 650 ng SOG/mg tissue/day. Maximum isthmic SOG secretion was approximately 30 ng SOG/mg tissue/day. These findings suggest that the oviduct is composed of discrete functional regions that provide support to gametes and developing embryos through the unique secretory characteristics of each region.

Sulfated Oviductal Glycoproteins in the Rabbit: Quantitation by Competitive Enzyme-Linked lmmunosorbent Assay1

The rabbit oviductal epithelium synthesizes and secretes a family of antigenically related, sulfated oviductal glycoproteins (SOG). Anti-SOG monoclonal antibodies (Mabs) were produced and two (Mab 1 and Mab 2) were selected for further characterization. Periodate oxidation of Western blots of oviductal fluid did not affect the binding of Mab 1 or Mab 2, thus suggesting that these antibodies recognized protein rather than carbohydrate epitopes on SOG. The specificity of Mab 1 was determined by Western blot analysis of tissues obtained from estrous rabbits and from the male rabbit reproductive tract. SOG was identified in tissue extracts of both the oviductal ampulla and isthmus. Cervix was the only non-oviductal tissue with which Mab 1 cross-reacted. Mab 1 was used to isolated SOG from whole oviductal fluid by immuno-affinity chromatography. Affinity-purified SOG and Mab 1 were used to develop a quantitative, SOG-specific, competitive enzyme-linked immunosorbent assay. This assay was used to quantify SOG in rabbit oviductal fluid collected during estrus and pseudopregnancy. SOG secretion during pseudopregnancy was resolved into two transient episodes of increased secretion. Maximum SOG secretion (X = 1039 +/- 199 micrograms/day) occurred within 48 h of the induction of pseudopregnancy. A second period of enhanced SOG secretion (X = 308 +/- 46 micrograms/day) occurred during the fifth and sixth days of pseudopregnancy. Baseline SOG secretion occurred during estrus at approximately 60% of maximum postovulatory secretion.

Differences between mature ovarian and oviductal oocytes: a study using the golden hamster

To investigate whether mature ovarian oocytes are physiologically identical with recently ovulated oocytes, hamster oocytes collected from ovaries approximately 1 h before ovulation were compared with the oocytes which had been in the oviduct for 3-4 h. Although these two groups of oocytes were at the metaphase of the second meiosis, there were quantitative differences between the two with respect to (i) sensitivity of cumulus matrix to hyaluronidase and spermatozoa (oviductal greater than ovarian), (ii) size of the perivitelline space (oviductal greater than ovarian), (iii) viscosity of the ooplasm (ovarian greater than oviductal), (iv) responsiveness to Ca2+-ionophore (oviductal greater than ovarian), and (v) time needed in completing meiosis (ovarian greater than oviductal). The most prominent difference was found in the zona pellucida. The zona of the oviductal oocyte was 'heterogeneous' in its optical density and had a stronger acrosome-reaction-inducing ability than that of the ovarian oocyte. When cultured in artificial media (e.g. F-10 medium with serum) for 4 h, the ooplasm of ovarian oocytes became like that of oviductal oocytes. However, their zonae remained unchanged. Zonae of ovarian oocytes became like those of oviductal oocytes only when they were exposed to ampullary and/or isthmic fluids. The zona-altering factors in the oviductal fluid (oviductal glycoproteins), which are apparently integrated into the native zona, may act to enhance the various functions of the zona.

Oviductal Glycoproteins Associated with the Pig Zona Pellucida: Immunocytochemical Localization in the Oviductal Epithelium

Oviductal Glycoproteins Associated with the Pig Zona Pellucida: Immunocytochemical Localization in the Oviductal Epithelium

Characterization of an Oviductal Glycoprotein Associated with the Ovulated Hamster Oocyte1

Hamster oviducts in culture incorporate [35S]-methionine into secretory proteins. One of these proteins is immunoprecipitated by a monoclonal antibody specific to an antigen found in oviductal oocytes but not in ovarian oocytes. This antigen, called oviductin, is progressively added to the oocyte during its transit through the oviduct. Oviductin migrates as a diffuse band with a molecular mass between 160 and 250 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing conditions. The electrophoretic behavior of this protein suggests the presence of polysaccharide side chains. Chemical deglycosylation causes a decrease in molecular mass and removes the antigenic determinant originally present on the glycoprotein. By using the radiation inactivation method, the molecular mass of the core protein has been found to be approximately 44 kDa. These results indicate that the oviduct is an actual site of synthesis of the oviductin. This glycoprotein contains a high proportion of sugar residues, which account for antigenic determinant recognized by the monoclonal antibody.

Immunocytochemical localization and determination of hormone-induced synthesis of the sulfated oviductal glycoproteins.

Secretory products of the oviduct provide part of the milieu for the critical events of fertilization and embryo development. Past work from this laboratory has indicated that three large sulfated glycoproteins can be isolated from rabbit oviductal fluid and are synthesized by oviductal epithelium incubated in vitro. These three glycoproteins are antigenically similar. This paper presents evidence for their localization within the oviductal tissue and their hormonal control of synthesis. Utilizing goat antiserum to these oviductal glycoproteins and the immunoglobulin-horseradish peroxidase bridge method, these macromolecules have been localized in the ampulla and isthmus of the oviduct. Ten days after ovariectomy an oviduct was removed for immunolocalization. The does were then given estradiol for the next 4 days and the second oviduct was removed. Oviducts treated with estradiol showed immunostaining of virtually all of the secretory granules within the secretory cells of the isthmus. While light level immunocytochemistry suggested the possibility of two populations of secretory granules within the ampulla because some of the granules did not show immunocytochemical staining, the more sensitive immunocytochemistry at the electron microscopic level showed staining of all granules of the ampulla and isthmus. Absorption of the antiserum with pure antigen prevented all staining. After ovariectomy and hormone withdrawal, most of the immunostaining was lost in the isthmus and virtually no staining in the ampulla was observed. Oviductal cell suspensions were made to evaluate incorporation of [35S] sulfate and [3H] leucine as a function of hormonal priming of the tissue. Estrogen-primed oviductal cells incorporated the sulfate and leucine into these specific glycoproteins.(ABSTRACT TRUNCATED AT 250 WORDS)

Induction of avidin in chickens infected with the acute leukemia virus OK 10.

The presence of avidin, a progesterone-dependent oviductal glycoprotein, was studied in viral tumors. Newborn chickens were infected with the acute leukemia virus OK 10, and the first tumors occurred within 2-3 weeks. Avidin was assayed using a [14C]biotin-binding method and radioimmunoassay. In the control chickens, avidin concentrations were less than 0.3 microgram/g in the plasma and less than 1.5 microgram/g in various tissues including the immature oviduct. In the OK 10 virus-infected chickens, no significant induction was observed during the acute infection or any time thereafter if no tumors were seen. In chickens that developed tumors, avidin concentrations were significantly increased in tumorous tissue only located in the mesenterium and occasionally in the oviduct. In tumors occurring elsewhere (kidney, ovary, muscle, testis, liver, colon) avidin concentrations were not elevated. Tumor-associated avidin had extraordinary biotin-binding capacity after treatment at +90 degrees C similar to the progesterone-dependent avidin, whereas antibody-binding properties suggested that tumor-associated avidin may have a somewhat altered antigenic structure.