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Background The objective of the present study was to define the morphological changes that occur in the epithelium of the isthmus oviduct of the sheep during the first few weeks of pregnancy. Materials: Isthmus oviducts were obtained from ovariectomized, estrous (day O) and pregnant (day 2, 3, 4, 6, and 16) ewes and analyzed using light and electron microscopy. Results The epithelium was low cuboidal in the isthmus from ovariectomized sheep, significantly increased in height at estrus, underwent an additional increase at day 3, and was significantly reduced by day 4 of pregnancy. Ciliated and nonciliated epithelial cells were present at all reproductive stages. Ciliated cells were always the predominant cell type. The percentage of ciliated and nonciliated cells did not significantly change at any stage examined. In addition, a third cell type, located adjacent to the basement membrane, was present. These “basal” cells were rounded in appearance and more frequently observed in the epithelial lining of ovariectomized ewes and after day 3 of pregnancy. The secretory organelles of nonciliated epithelial cells obtained from ovariectomized ewes were poorly developed. In estrous ewes, the Golgi apparatus in nonciliated cells consisted of stacked cisternae that atrophied later in pregnancy. The most striking alterations in the secretory apparatus occurred in ciliated cells. Two populations of ciliated cells were observed at estrus-one with abundant, membrane bound supranuclear granules and one without granules. The granules in ciliated cells were rare after day 3 of pregnancy and no evidence of secretion was ever observed. Large lipid droplets were present in the cytoplasm of ciliated cells at estrus. Lysosomes were abundant in nonciliated cells by day 16 of pregnancy. Regularly arranged microvilli were present in nonciliated cells obtained from ovariectomized, estrous, and pregnant ewes. No appreciable change in nuclear shape or chromatin content was observed at any reproductive stage examined. The ultrastructural characteristics and relative abundance of other cytoplasmic organelles such as the RER, SER, mitochondria, and glycogen varied slightly during pregnancy in ciliated and nonciliated cells. Conclusions These data show that the epithelial lining of the isthmus oviduct in the sheep undergoes subtle, yet distinctive, structural changes during the first few weeks of pregnancy. Anat. Rec. 247:368–378, 1997. © 1997 Wiley-Liss, Inc.

Data from our laboratory have shown that an estrogen (E2)-dependent M(r) 90,000-92,000 protein and its mRNA are synthesized and expressed in abundant amounts at estrus from the fimbria and ampulla, not isthmus, oviduct of the sheep. Immunocytochemical studies have shown that the M(r) 90,000-92,000 protein is contained in apical secretory granules of oviduct epithelial cells. The objective of this study was to determine whether the mRNA for the E2-dependent oviduct protein was localized and compartmentalized in similar manner. Fimbria, ampulla, and isthmus oviducts obtained from estrous ewes were flash frozen in liquid nitrogen, cryosectioned, fixed in 4% paraformaldehyde, hybridized with digoxigenin (DIG)-labeled oviduct-specific riboprobes, incubated in anti-DIG antibodies conjugated with alkaline phosphatase, and developed in color substrate. Oviduct protein-specific transcripts were localized to basal perinuclear compartments and, surprisingly, at sites distant from the nucleus in the apical cytoplasm of epithelial cells in the fimbria and ampulla. No specific reaction product was observed in the underlying mucosa or smooth muscle layers. Oviduct protein mRNA was contained predominantly in the apical cytoplasm of epithelial cells at the free margins of mucosal folds and in the basal regions of cells located at the crypts of longitudinal folds. No reaction product was present when sections of the fimbria and ampulla oviduct of estrous ewes were incubated in sense riboprobe to the oviduct protein. In addition, when sections of the isthmus oviduct obtained from estrous ewes or fimbira and ampulla oviducts from long-term ovariectomized ewes were hybridized with antisense riboprobes no specific reaction product was detected. Electron microscopy of oviduct protein mRNA containing areas revealed the presence of secretory granules, rough endoplasmic reticulum (RER) and Golgi in the apical cytoplasm, and RER in the basal regions of epithelial cells. These data show that the mRNA encoding an E2-dependent oviduct-specific protein is distributed in epithelial cells at perinuclear foci and at sites distant from the nucleus, which are also the sites of protein localization and protein synthesizing organelles, implying translation at unique cytoplasmic foci.

This study was undertaken (1) to devise a method of inducing multiple follicular development and subsequent ovulation in the Djungarian or Siberian hamster (Phodopus sungorus) and (2) to assess the quality of ovulated oocytes collected from PMSG/hCG treated animals in comparison to naturally ovulating animals. Hamsters (4–5 weeks; n = 70) received 5 IU PMSG followed 50–52 hr later by 10 IU hCG. Ovulated oocytes were collected 14–20 hr after hCG injection. Ovulated oocytes were flushed from oviducts of cycling animals (7–12 weeks; n = 30) exhibiting two consecutive estrous cycles. Oocytes were fixed and subjected to triple fluorescence immunostaining using anti-tubulin antibodies, fluorescein phalloidin, and Hoechst 33258. The mean number of ovulated oocytes collected from cycling animals was 4.8 ± 0.4 (range 1–7). Ovulation occurred in 73% of the PMSG/hCG-stimulated animals. The mean number of oocytes ovulated from stimulated animals was 9.2 ± 0.8 (range 0–22). The ovaries of animals that did not ovulate or that ovulated few oocytes did respond to PMSG, as indicated by the presence of multiple follicular development and pre-ovulatory stigmata. There was no evidence of a polar body in ovulated oocytes collected from PMSG/hCG-treated or cycling animals, indicating that oocytes were arrested in meiosis I. In the majority (80%) of ovulated oocytes from PMSG/hCG-treated and cycling animals, cortically placed chromosomes were aligned on a metaphase plate equidistant from a bipolar spindle. Sparse f-actin staining was observed in the region of the ooplasm surrounding the chromosomes. As the interval between hCG injection and the time of collection increased, chromosomes lost their proper alignment and migrated away from the cortex of the oocyte concomitant with a disruption of spindle integrity. This collapse of proper chromosome alignment and disruption of spindle architecture also characterized aging oocytes collected from cycling animals. These data show that in the Djungarian or Siberian hamster (Phodopus sungorus), (1) there is individual animal variation in responsiveness to hCG following PMSG treatment, (2) there are no cytological differences in the quality of oocytes collected from hormonally treated animals when compared to cycling animals, and (3) oocytes are ovulated in meiosis I. © 1995 Wiley-Liss, Inc.

Oviducts and uteri were obtained from cats at estrus and up to 14 days after coitus. Progesterone binding sites were estimated by Scatchard analysis, serum levels of progesterone were determined by radioimmunoassay, and key morphological parameters of the lining of the epithelium of the oviduct and the deep uterine glands were quantitated. Progesterone cytosol binding was hightest at estrus, significantly depleted by day 4, and continued a gradual decline out to day 14 in both tissues. Nuclear binding was markedly elevated at day 4; with time the binding decreased until by day 14 it was equal to or lower than detected at estrus. The initial depletion of cytosol binding and rise in nuclear binding coincided with the initial rise in serum progesterone levels. Within the oviduct, the increase in nuclear binding coincided with the onsdet of regressive changes within the epithelium. These changes included atrophy, deciliation, and apotosis. Within the uterine glands, the increase in nuclear binding coincided with the onset of the progestational response. This response included cell hypertrophy, glycogen synthesis and deposition, and with longer exposure to elevated levels of progesterone, the disappearance of these glycogen deposits. These results clearly illustrate the immediate effect of low levels of progesterone on the subcellular compartmentalization of the progesterone receptor system, and the down regulation of the total receptor system when target tissues are exposed to a continuous elevation of serum progesterone. These result also illustrate that similar changes in receptor compartmentalization induce different morphological responses within target tissues from the same animal.