Immunogold Localization of Porcine Oviductal Secretory Proteins within the Zona Pellucida, Perivitelline Space, and Plasma Membrane of Oviductal and Uterine Oocytes and Early Embryos1

Abstract

The objectives of the present study were to develop an antibody probe to the porcine estrogen-dependent oviductal glycoproteins and to determine, by use of immunogold electron microscopy, whether these glycoproteins become associated with oviductal and uterine oocytes and early embryos. Polyclonal antibody, prepared using the M(r) 75,000-85,000 glycoprotein, separated from other proteins by two-dimensional SDS-PAGE, specifically recognized all three estrogen-dependent glycoproteins (acidic 75,000-85,000 M(r); acidic 100,000 M(r); basic 100,000 M(r)). In ampullary tissue collected from ovariectomized and estrogen-treated gilts and from gilts at Day 1 of estrus, gold particles were clustered over putative secretory granules restricted to the apical region of secretory epithelial cells. While follicular oocytes did not react with immunoreactive colloidal gold, oviductal and uterine unfertilized oocytes were found to be densely and uniformly labeled by colloidal gold throughout the zona pellucida, associated with flocculent material in the perivitelline space, and associated with microvilli and vitelline membrane. Similarly, in oviductal (1-4-cell) and unhatched uterine (4-cell/blastocyst) embryos, colloidal gold particles were distributed throughout the zona pellucida, heavily associated with flocculent material in the perivitelline space, and associated with the plasma membrane of the blastomeres. Immunoreactive colloidal gold remained detectable within Day 7 hatched uterine embryos, but not with embryos from later days. These results further support the proposal that porcine estrogen-dependent oviductal glycoproteins are released into the oviductal lumen, become associated with oviductal and uterine oocytes and early embryos, and are retained by oocytes and early embryos in the uterus.

Oviduct-specific glycoprotein and heparin modulate sperm–zona pellucida interaction during fertilization and contribute to the control of polyspermy

“…6 A and B) and, once fertilization and blocking of polyspermy have occurred, the factor is either partially and slowly removed from the zona during its transit toward the uterus (Fig. 6D) or incorporated through the perivitelline space to the plasma membrane, as has been shown in pigs for the OGP (35). The cumulus cells accompanying the oocyte from the follicle are physiologically removed shortly after ovulation, although their presence does not hamper the OGP binding to the ZP (36).…”

Amount of Hyaluronan Produced by Mouse Oocytes and Role of Hyaluronan in Enlargement of the Perivitelline Space

“…On the other hand, it is known that proteins and HA are present in perivitelline spaces of oocytes before ovulation [11][12][13][14]39] and that secretions from the oviducts [40][41][42][43][44] are also present after ovulation. When the perivitelline space decreases in zize, the quantities of substances in the perivitelline space also decrease.…”

Localization of Immunoreactive Gonadotropin-releasing Hormone and Relative Expression of Its mRNA in the Oviduct During Pregnancy in Rats

“…Involvement of oviductal epithelium of the isthmus in the secretion of an oviduct-specific high molecular mass glycoprotein during the estrous cycle in rats has been reported (Abe 1996). Reports regarding the synthesis and release of proteins/glycoproteins by oviductal epithelium in other mammals like mice (Zhang and Martin-DeLeon 2003), pigs (Buhi et al 1993(Buhi et al ,1997, cows (Gerena and Killan 1990), and baboons (Boice et al 1990) also exist. These proteins/glycoproteins secreted by the oviductal epithelium were found to associate with either zona pellucida of the oviductal ova or the early embryos (Kan et al 1993), implicating their paracrine role in fertilization and early embryonic development.…”