Estradiol induces E-cadherin degradation in mouse uterine epithelium during the estrous cycle and early pregnancy

Mouse uterine epithelium is a tissue that undergoes cyclic endocrine-regulated cell dissociation and regeneration. It shows a dramatic cell loss following normal estrus. If pregnancy ensues, cell loss is averted during the first 2.5–3.5 days. However, this is followed by a precipitous loss of basal-lateral cell adhesion and apoptosis in preparation for blastocyst invasion. By comparing epithelia isolated by protease treatment, we show that a reduction of lateral cell adhesion is a primary event in these instances of normal tissue loss. It was readily induced in ovariectomized adult and immature mice by injections of estradiol (E₂), and to some extent also by progesterone (P₄). The reduction of lateral adhesion induced by including ethylene glycol-bis (β-aminoethyl ether)-N,N,N′,N′-tetraacetic acid (EGTA) in the isolation medium mimicked and was additive to the effect of E₂ injection. However, the E₂ effect was different in not being prevented by adding Ca²⁺. The E₂ effect also was mimicked by the action on isolated epithelium of monoclonal antibody against the calcium-dependent cell adhesion molecule, E-cadherin, suggesting that inactivation of E-cadherin was induced by E₂. In detergent extracts of estrous and metestrous epithelium there was an increase in 80-kDa extracellular domain of E-cadherin relative to the intact 120-kDa molecule. The loss of adhesion between 3.5 and 4.5 days of pregnancy was associated with a loss of both intact membrane-associated 120-kDa E-cadherin and cleavage products. Cleavage of 80-kDa E-cadherin was uniquely induced by E₂ in ovariectomized adult and immature mice; P₄ was without effect. The cleavage of E-cadherin correlated with increased basal accumulation of E-cadherin antigen in estrous and E₂-injected mice and a loss of both basal and lateral antigen at 4.5 days of pregnancy. Only the E-cadherin antigen within junctional complexes appeared unaffected. The data are consistent with the hypothesis that the cyclic and pregnancy-dependent disruption of uterine epithelial integrity are promoted by E₂-dependent modification of E-cadherin, including its extracellular cleavage. © 1996 Wiley-Liss, Inc.     

Lymphocyte number and distribution in the rat uterine epithelium during estrous cycle and early pregnancy

Summary The number of intraepithelial lymphocytes (IEL) in the luminal and glandular epithelium of the uterus of virgin rats was analysed in diestrus, proestrus and estrus, and in nulliparous rats on days 5, 7 and 9 of pregnancy. IEL number was calculated either with respect to the number of epithelial cells or to the length of epithelium section. It was found that in diestrus, the number of IEL was, on average, 3.7 per 100 luminal epithelial cells or 6.7 per 1 mm of epithelium section, whereas in proestrus, it decreased to 0.9 and 1.2 IEL, respectively. On day 5 of pregnancy (before implantation) the number of IEL decreased further to 0.45 per 100 luminal epithelial cells or 0.9 per 1 mm of epithelium. On days 7 and 9 of pregnancy, IEL number further decreased in implantation sites, whereas in interimplantation sites it remained at the level calculated for day 5 of pregnancy. The population of uterine IEL consisted of small (82–99%) and large (1–18%) lymphocytes. In all stages of the estrous cycle, IEL occurred with a frequency of 68–87% in the basal region, 8–20% in the middle region and 4–12% in the apical region of the luminal epithelium width.     

The ultrastructure of the uterine epithelium of the pig during the estrous cycle and early pregnancy

Summary In the compound eye of the moth Antheraea polyphemus, three types of visual pigments were found in extracts from the retina and by microspectrophotometry in situ. The absorption maxima of the receptor pigment P and the metarhodopsin M are at (1) P 520–530 nm, M 480–490 nm; (2) P 460–480 nm, M 530–540 nm; (3) P 330–340 nm, M 460–470 nm. Their localization was investigated by electron microscopy on eyes illuminated with different monochromatic lights. Within the tiered rhabdom, constituted of the rhabdomeres of nine visual cells, the basal cell contains a blue-and the six medial cells have a greenabsorbing pigment. The two distal cells of most ommatidia also have the blue pigment; only in the dorsal region of the eye, these cells contain a UV-absorbing pigment, which constitutes a portion of only 5% of the visual pigment content within the entire retina. The functional significance of this distribution is discussed.     

Chronic utero-ovarian vein catheterization with subsequent occlusion prolongs the estrous cycle and changes electromyographic activity in the myometrium of ewes

The objective of our study was to determine the effect of chronic utero-ovarian vein catheterization in ewes on estrous cycle length, plasma progesterone (P) concentration, and myometrial electromyographic activity. Cyclic ewes with inferior vena cava catheters were used as controls. Estrus was synchronized in ten ewes and 10 to 12 d following estrus, the ewes were anesthetized, fitted with myometrial electromyograph leads and with utero-ovarian vein (n = 5) or inferior vena cava (n = 5) catheters. After surgery, ewes returned to estrus as expected (16 to 18 d interestrus interval). The second cycle of four of five ewes with utero-ovarian vein catheters were prolonged (40 to 58 d). The inferior vena cava catheterized ewes had normal length second cycles. Plasma P concentrations reflected the estrous cycles: low ( 0.05).     

Antioxidant capacity of follicular fluid in relation to follicular size and stage of estrous cycle in buffaloes

The present study was designed to evaluate the changes in the concentrations of different antioxidants, such as glutathione (GSH), glutathione reductase (GR), superoxide dismutase (SOD), and catalase (CAT), in the follicular fluid collected from different follicular size categories in relation to stage of estrous cycle in buffaloes. In addition, malondialdehyde (MDA) as an indicator for lipid peroxidation was also estimated. Fifty pairs of buffalo ovaries were collected from a local slaughterhouse. Based on ovarian structures, the cycle was divided into follicular and luteal phase. The follicles on each pair were classified into three groups; small (≤3 mm), medium (4-9 mm) and large (≥10 mm). The concentrations of SOD, CAT, GSH, and GR in the follicular fluid of each group as well as MDA were estimated. Results indicated that there was a significant decrease (P < 0.05) in the average numbers of small follicles obtained at the follicular phase than those obtained at the luteal phase of the cycle. However, the mean numbers of the large sized follicles was significantly increased (P < 0.05) in the follicular phase than in the luteal phase. Large follicles obtained at the luteal phase had a significantly higher (P < 0.05) concentration of GSH than that obtained from small ones. A significant (P < 0.05) effect of follicular size on GR concentrations was observed. The concentration of SOD tended to be higher in large follicles obtained at the follicular phase than that collected at the luteal phase (56.7 ± 3.7 vs. 28.1 ± 6.7 U/mL, respectively). On the contrary, a significantly higher concentration (P < 0.05) of SOD was recorded in small follicles as compared with medium and large follicles collected at the luteal phase. CAT concentrations did not significantly differ among different follicular sizes between follicular and luteal phases as well as within each phase. Malondialdehyde concentration was significantly decreased (P < 0.05) in the follicular fluid obtained from small follicles collected at the follicular phase compared with those obtained at the luteal phase. In conclusion, the present study showed that the concentrations of enzymatic antioxidants except for CAT vary according to the follicle size and the stage of the estrous cycle suggesting their possible role in the process of follicular development during estrous cycle in buffaloes.     

The estrous cycle in the dog: a review

Clinical, morphologic and hormonal changes throughout the estrous cycle in the dog are discussed in relation to the well established stages in this species — proestrus, estrus, metestrus and anestrus. Although most efforts in the past have been devoted to the period of reproduction, events throughout the remaining months of the cycle are presented to characterize the dog as a monocyclic animal in contrast to polycyclic species.     

Characterization and distribution of cholinesterase activity in mouse uterine horns: changes in estrous cycle

1. Both butyrylcholinesterase (BChE) and acetylcholinesterase (AChE) are present in the mouse uterus, BChE being more abundant.2. Their molecular forms were sequentially solubilized by different extraction media obtaining three ChE fractions whose specific activity was different, depending on the stage of the estrous cycle: hydrosoluble (estrous: 75.5 ± 6.6 and diestrous: 47.9 ± 8.7 μU/mg prot); detergent-soluble or amphiphilic (estrous 26.6 ± 2.4 and diestrous 14.7 ± 3.3 mU/mg prot.), and high ionic strength-soluble (estrous: 18.7 ±4.2 and diestrous 12.8 ± 1.2 mU/mg prot.).3. Histochemical procedures demonstrated a different distribution for both ChE activities. AChE was found in nerves next to smooth muscle cells of the circular layer and blood vessels, while BChE was concentrated in the longitudinal stratum surrounding the smooth muscle cells. Under the predominance of progesterone, BChE was also found in the endometrial glands.4. Maximal contractions evoked by the addition of ACh to the isolated organ bath were concentration dependent and greater in estrous than in diestrous. Nevertheless the difference at the two stages of the estrous cycle disappeared when contractions were normalized to smooth muscle cross-sectional area.5. BChE but not AChE inhibition augmented maximal contractions elicited by ACh in longitudinal but not in circular smooth muscle.6. The effect of BChE inhibition on the contractile force developed was greater at lower concentrations of ACh and did not depend on the stage of the estrous cycle.     

Peripheral inhibin levels in relation to climatic variations and stage of estrous cycle in buffalo (Bubalus bubalis )

The present study investigated the peripheral plasma inhibin levels in relation to 1) the stage of estrous cycle and the effect of climatic variations. Blood samples were collected from cyclic buffalo (n=5) once daily for 32 consecutive days during the tropical hot humid (summer) and cold (winter) seasons. Estrus was recorded by parading a vasectomized bull as well as by plasma progesterone determination. In the winter season, peripheral inhibin concentrations which were lowest (0.35 +/- 0.02 ng/ml) during the mid-luteal phase of estrous cycle (Day 6 to Day 14, Day 0 = day of estrus) increased significantly (P < 0.02) to 0.47 +/- 0.04 ng/ml during the late luteal phase (Day -4 to Day -2) and then further to 0.52 +/- 0.03 ng/ml (P< 0.02) during the periestrus phase (Day -1 to Day 1). Inhibin concentrations then decreased significantly (P < 0.02) to 0.40 +/- 0.03 ng/ml during the early luteal phase (Day 2 to Day 5). In the summer season the differences in peripheral inhibin concentrations among different phases of estrous cycle were found to be nonsignificant. A comparison of the circulating inhibin concentrations between the two seasons indicated that inhibin concentrations were significantly higher in the late luteal phase (P < 0.01) and periestrus phase (P < 0.05) during the winter season compared with corresponding periods during the summer season. The present study suggests that peripheral inhibin concentrations change in the estrous cycle during cooler breeding season and that environmental heat stress can cause a reduction in peripheral inhibin concentrations.     

Changes in the distribution of ZO-1, occludin,and claudins in the rat uterine epithelium during the estrous cycle

During the estrous cycle, the endometrium epithelium experiences marked cellular structural changes. For fertilization to proceed, maintenance of an adequate uterine environment by ovarian hormones is essential. Epithelial cells lining the uterine lumen are associated with each other by tight junctions (TJs), which regulate the passage of ions and molecules through the paracellular pathway. The aim of the present study was to assess by confocal immunofluorescence the distribution pattern of the TJ proteins ZO-1, occludin, and claudins 1–7 in the rat uterus during the estrous cycle. Our results reveal that on proestrus, the day when mating takes place, ZO-1, occludin, and claudins 1 and 5 are located in the TJs, while claudins 3 and 7 display a basolateral distribution. In contrast, on metestrus day, when no sexual mating occurs and the uterine lumen is devoid of secretions, none of these proteins were detected in the TJ region, and only a diffuse cytosolic staining was observed for some of the proteins. On estrus and diestrus days, an intermediate situation was encountered, since ZO-1 localized in the TJs, whereas occludin was no longer detectable in the TJs. The distribution of claudins during these stages varied from the lowermost portion of the basolateral membrane to its apex. In conclusion, the results show that the protein composition of TJs present in the luminal epithelial cells of the uterus changes during the different days of the estrous cycle, and suggest that the expression of TJ proteins participates in providing an adequate environment for a successful fertilization.This work was supported by grants PAPIIT (IN210902, IX228504) and PAIP (6190-08) from the National Autonomous University of Mexico (UNAM), and by grants G34511-M and 37846-N from the Mexican National Council on Science and Technology (CONACYT).     

Evolution of the flagellar waveform of ram spermatozoa in relation to the degree of epididymal maturation.

Motility and flagellar movement of ram spermatozoa along the epididymis were analysed in vitro. From the caput to the cauda of the epididymis, the percentage of motile and progressive spermatozoa increases. No flagellar bending was observed in spermatozoa from the testis or the epididymal anterior caput. When spermatozoa reached the distal caput of the epididymis, a static curvature, associated with an initiation of the flagellar beating, appeared on the flagella. This curvature normally disappeared during epididymal transit. Its disappearance was associated with an increase in the flagellar beat efficiency. Our results suggest that the initiation of motility is related to two mechanisms involving: (1) the presence of a transient static curvature, and (2) the establishment of a symmetric regular beating of the flagellum.     

Diffusion barriers in the vaginal epithelium during the estrous cycle in guinea pigs

Summary In the present study the permeability barriers of the multilayered vaginal epithelium were examined using tracer perfusion techniques, freeze-fracture and thin sectioning. During diestrus and proestrus the upper layers of mucified epithelial cells exhibit tight-junctional belts, which restrict tracer molecules such as lanthanum and horseradish peroxidase. When the highly mucified cells begin to degenerate toward the end of proestrus the underlying epithelium is already keratinized as typical for estrus. The keratinized epithelial cells have a tight-junctional network that joins the basal plasma membranes with the apical membranes of subjacent cells and blocks paracellular diffusion of the tracer molecules. During conversion of the cornified epithelium to a mucified epithelium in metestrus the intercellular space of the epithelium is stained by tracer molecules even though tight-junctional belts can be observed.These results indicate that during cyclic changes of the vaginal epithelium tight junctions can, in general, be considered for the restriction of paracellular diffusion. In metestrus, however, junctions become functionally leaky although they remain morphologically intact.Intercellular lipids, which are normally common in cornified epithelia, are extremely rare and cannot constitute an effective barrier to diffusion in the vagina of the guinea pig. The significance of a strategy that bases the regulation of the permeability on tight junctions rather than on intercellular lipids is discussed.     

Ultrastructural changes in the oviductal epithelium of Merino ewes during the estrous cycle

Isthmic and ampullary oviductal epithelia sampled from Merino ewes at days -1, 1, 3, and 10 of the estrous cycle (estrus = day 0) were studied by scanning and transmission electron microscopy after fixation by vascular perfusion. Secretory cells, ciliated cells, and lymphocytelike basal cells were observed in both isthmic and ampullary epithelium at all stages of the estrous cycle studied and their ultrastructural features were analyzed. Synthesis of lamellated secretory granules occurred in the ampullary secretory cells during the follicular and early luteal phases, and their contents were released by exocytosis into the oviductal lumen during the luteal phase. Granule release was associated with nucleated apical protrusion of these cells into the oviductal lumen. No such secretory activity was displayed by isthmic secretory cells even though a few cells contained nonlamellated granules. Apocrine release of apical vesicles and accompanying cytoplasmic material from apical protrusions of ciliated cells occurred in the isthmus around estrus but not in the ampulla. This unexpected feature has not previously been reported in any other mammal. Dendritic basal cells were distinguished in the lower part of the epithelium by their heterochromatic nuclei, electron-lucent cytoplasm, and lack of attachment zones. No migration of basal cells was observed, and their ultrastructural features were similar in the ampulla and isthmus and at all stages of the estrous cycle examined. The function of these lymphocytelike cells in the epithelium is uncertain, but the presence of phagocytic bodies and lysosomes in 20% of them may indicate a phagocytic role.