Ovine uterine morphogenesis: histochemical aspects of endometrial development in the fetus and neonate

Uterine tissues obtained from fetal (d 60 to term; n = 17; d 0 = day of mating) and neonatal (n = 9; d 0 = birth) lambs were subjected to alcian blue-8GX and fluorescein isothiocyanate (FITC)-lectin histochemistry to determine if alcianophilic properties of the epitheliomesenchymal interface (EMI) changed during endometrial morphogenesis and to characterize distribution of binding sites for seven FITC-lectins during uterine development. Neonatal lambs were subjected to bilateral ovariectomy and unilateral hysterectomy (BOHX; n = 3) or unilateral ovariohysterectomy (UOHX; n = 3) on d 0. Remaining tissues were recovered on d 14. Procedures allowed within-animal comparisons of endometrial responses and assessment of the role of the ovary in endometrial morphogenesis. Uteri also were obtained from three intact neonatal lambs by hysterectomy (d 14, d 15 and d 26). Alcianophilic properties of the EMI characteristic of polyanionic glycosaminoglycans (GAG) changed with onset of endometrial remodelling after fetal d 60 and were characterized by loss of EMI alcianophilia at or above .3 M MgCl2 and at low pH. Alcianophilic properties of the neonatal endometrium suggested restabilization of lumenal EMI and destabilization of the EMI in developing endometrial glands. Five of seven FITC-lectins bound to both fetal and neonatal uterine tissue. Tissues from UOHX, BOHX and intact ewes were indistinguishable histochemically. Data provide evidence of a role for GAG in ovine endometrial morphogenesis, ovary-independent initiation of endometrial glandular development, and illustrate potential uses of FITC-lectin conjugates in studies of ungulate uterine tissues.     

Crb1在小鼠生后不同发育阶段子宫组织中的表达

应用HE染色方法和免疫荧光组织化学技术对小鼠生后不同发育阶段子宫组织结构的发育以及极性调控蛋白Crb1的定位表达进行了研究,结果显示,随个体发育,子宫管腔及子宫腺腔不断扩大,子宫内膜上皮持续增厚、固有层内子宫腺逐渐发达;4周龄时子宫腔面出现纤毛,随发育进程越来越发达;在各发育阶段的小鼠子宫组织中均有Crb1的表达,随个体发育表达呈现先增强后减弱的趋势,在8周龄时达到最强;Crb1主要定位于子宫内膜上皮细胞和子宫腺上皮细胞的胞膜和胞质,提示Crb1可能与小鼠子宫内膜上皮细胞和腺上皮细胞的极性建立和维持有关。     

Neonatal porcine endometrial development and epithelial proliferation affected by age and exposure to estrogen and relaxin

In the pig, temporospatially regulated proliferation of uterine luminal (LE) and glandular (GE) epithelium between birth (postnatal day=PND 0) and PND 15 is essential for success of endometrial development. Exposure of gilts to estrogen (E) or relaxin (RLX) during this period disrupts uterine development, and neonatal E exposure can compromise adult uterine function. Neonatal uterotrophic effects of E and RLX, administered for 2 days beginning on PND 12, can be inhibited with the antiestrogen ICI 182,780 (ICI) indicating crosstalk between RLX and E signaling systems. Here, objectives were to determine effects of: (study 1) neonatal age and (study 2) exposure to E, RLX, and ICI on porcine neonatal uterine histoarchitecture and patterns of epithelial cell proliferation as reflected by proliferating cell nuclear antigen labeling index. In study 1, uteri were obtained on PND 0, 3, 6, 9, 12 and 15. Glandular epithelium, absent at birth, was observed by PND 3. Overall, epithelial labeling index increased from birth to PND 3, declined from PND 6-9 in LE and GE, and increased to PND 15 in GE. In study 2, uteri were collected on PND 14 after administration of vehicle, E, or RLX for 2 days, or following pretreatment with ICI. Alone, E was uterotrophic and adenogenic and increased labeling index in both LE and GE. Both RLX and ICI increased proliferation in GE. Effects of E and RLX were attenuated by ICI, providing further support for crosstalk between these signaling systems in the developing neonatal porcine endometrium.     

Neonatal estradiol exposure alters uterine morphology and endometrial transcriptional activity in prepubertal gilts

Porcine endometrial development between birth (postnatal day = PND 0) and PND 56 involves differentiation of glandular epithelium (GE) from luminal epithelium (LE) and estrogen receptor-alpha (ER) expression. Juvenile ER architecture evolves after birth, as stroma and nascent GE first express ER. Mature ER architecture is evident after PND 30, when stroma, GE and LE are ER-positive. When administered during discrete periods between PND 0 and 56, effects of estradiol-17beta valerate (EV) on the neonatal porcine uterus relate to endometrial ER architecture. Transient EV exposure from birth reduces embryo survival in pregnant adult gilts. Effects of EV, administered as juvenile endometrial ER architecture develops (P1, PND 0-13), or after mature ER architecture is established (P2, PND 42-55), were evaluated in uteri from gilts treated with corn oil or EV in P1 or P2 and hysterectomized on PND 100 without additional steroids (NSt), on PND 102 after EV on PND100-101 (EV2), or on PND 117 after EV2 followed by progesterone on PND 102-116 (EP). Neonatal EV reduced uterine weight (P < 0.02), size (P < 0.01), luminal protein content (P < 0.07), and percent incorporation of 3H-leucine into nondialyzable endometrial products in vitro (P < 0.01). Group (NSt, EV2, EP) -specific treatment effects detected for endometrial ER, progesterone receptor, uteroferrin, and/or retinol binding protein mRNA levels were frequently related to period (P1,P2). Results support the idea that estrogen-sensitive postnatal organizational events, including those defined, in part, by endometrial ER architecture, are likely components of genetic and epigenetic programs governing uterine morphogenesis and ontogeny of endometrial function in the pig.     

Interrelationships among conceptus size, uterine protein secretion, fetal erythropoiesis, and uterine capacity

The interrelationships among d-11 conceptus size, d-105 placental weight, placental efficiency (the ability of the placenta to support fetal growth and development), fetal erythropoiesis, and uterine capacity were examined in 1/2 Meishan, 1/2 White crossbred gilts that were unilaterally ovariohysterectomized at 90 to 100 d of age. In Exp. 1, gilts were mated after at least one normal estrous cycle and then slaughtered at 105 d of gestation and number of fetuses and CL, placental weights, fetal weights, hematocrits, fetal plasma iron, and fetal plasma folate were measured. In Exp. 2, gilts were mated and plasma progesterone was measured on d 2 and 3 of gestation. On d 11, the length of the remaining uterine horn was recorded and the uterine horn was flushed with minimal essential medium. Number of CL, conceptus number, conceptus diameters, and total uterine flush retinol-binding protein (tRBP), acid phosphatase (tAP), and folate-binding protein (tFBP) were measured. Gilts were mated again and slaughtered at 105 d of pregnancy and the same traits measured in Group 1 were recorded. Plasma progesterone concentrations on d 2 and 3 were correlated with average conceptus diameter on d 11 (r = 0.60, P < 0.01, for each day). In contrast, tRBP (r = 0.49, P < 0.01), tAP (r = 0.53, P < 0.01), and tFBP (r = 0.51, P < 0.01) in uterine flushings on d 11 were only correlated with d-3 plasma progesterone concentrations. No correlations between d-11 average conceptus diameter or d-11 uterine length with d-105 uterine capacity were observed. Uterine capacity was negatively correlated with placental weight, fetal weight and fetal hematocrit (r = -0.36, P < 0.01; r = -0.44, P < 0.01; r = -0.32, P < 0.01; respectively). Hematocrits were correlated with fetal plasma iron (r = 0.50, P < 0.01) and folates (r = 0.44, P < 0.01). Hematocrit, plasma iron, and plasma folate were each correlated with residual fetal weights after adjusting for placental weight (a measure of placental efficiency), and accounted for 11% of the variation in this trait. These data suggest that conceptus diameter and uterine protein secretion on d 11 may be influenced by the onset of progesterone secretion by the CL, but do not support an influence of conceptus growth during early pregnancy on uterine capacity. These results also suggest that reducing placental and fetal weights will likely result in increased uterine capacity.     

Ovine uterine morphogenesis: effects of age and progestin administration and withdrawal on neonatal endometrial development and DNA synthesis

To determine effects of age and administration and withdrawal of a synthetic progestin (P) on endometrial development and DNA synthesis, ewe lambs were ovariectomized on d 0 (birth) and assigned to one of four groups (n = three/group) that provided (by means of hemihysterectomy) the following uterine tissue types: 1) d 0 control, 2) d 13 control, 3) d 26 control, 4) d 13 after 13 d exposure to P (13P) and 5) d 26 after P exposure from d 0 to 13 (26P). Uterine tissues were processed for histology or explanted with [methyl-3 H] thymidine for autoradiography. Labeling index (LI) was determined for stroma and epithelium in caruncular and intercaruncular endometrial areas and for lumenal and glandular epithelium in uteri with glands. Endometrial glands were absent on d 0, evident at d 13 and well developed by d 26. Day 0 LI was greater (P less than .05) in caruncular than in intercaruncular areas, and greater in stromal than in epithelial tissues. Relationships were reversed in d 13 endometrium (day X endometrial area, P less than .07). Caruncular stromal LI was greater on d 0 than later (P less than .02), whereas intercaruncular epithelial LI was greater after d 0 (P less than .05), but decreased from d 13 to 26 (P less than .05). Glandular epithelial LI was higher on d 13 than on d 26 (P less than .03). Administration of P inhibited endometrial gland development and suppressed d 13P intercaruncular LI (P less than .05). Withdrawal of P was followed by endometrial gland development and increased (P less than .01) intercaruncular epithelial LI in d 26P uteri. Ovary-independent initiation of endometrial gland development involves age- and region-specific alterations in DNA synthesis and could involve negative control.     

Origin of enzootic intranasal tumor in the goat (Capra hircus): a glycohistochemical approach

Enzootic intranasal tumor (EIT) appears glandular in type and has recently been classified as an adenocarcinoma of low malignancy. The aim of this study was to characterize the secretion of surface glycoconjugates (GCs) in EIT and in normal respiratory and olfactory mucosae of the goat by means of conventional and lectin histochemistry, in order to shed light on the histogenesis of EIT. Morphologic and ultrastructural investigations showed two growth types of EIT: i.e., tubular and papillary patterns. Conventional histochemistry revealed the presence of neutral and carboxylated GCs in the olfactory glands and in the tubular part of EIT, as well neutral and sulphated GCs in the respiratory glands and in the papillary part of EIT, suggesting that the papillary pattern tumor arises from the respiratory glands, whereas the tubular portion of EIT arises from the olfactory glands. Lectin histochemistry gave further information on the expressed GCs.     

Structure and Steroidogenesis of the Placenta in the Antarctic Minke Whale (Balaenoptera bonaerensis)

There are few reports describing the structure and function of the whale placenta with the advance of pregnancy. In this study, therefore, the placenta and nonpregnant uterus of the Antarctic minke whale were observed morphologically and immunohistochemically. Placentas and nonpregnant uteri were collected from the 15th, 16th and 18th Japanese Whale Research Programme with Special Permit in the Antarctic (JARPA) and 1st JARPA II organized by the Institute of Cetacean Research in Tokyo, Japan. In the macro- and microscopic observations, the placenta of the Antarctic minke whale was a diffuse and epitheliochorial placenta. The chorion was interdigitated to the endometrium by primary, secondary and tertiary villi, which contained no specialized trophoblast cells such as binucleate cells, and the interdigitation became complicated with the progress of gestation. Furthermore, fetal and maternal blood vessels indented deeply into the trophoblast cells and endometrial epithelium respectively with fetal growth. The minke whale placenta showed a fold-like shape as opposed to a finger-like shape. In both nonpregnant and pregnant uteri, many uterine glands were distributed. The uterine glands in the superficial layer of the pregnant endometrium had a wide lumen and large epithelial cells as compared with those in the deep layer. On the other hand, in the nonpregnant endometrium, the uterine glands had a narrower lumen and smaller epithelial cells than in the pregnant endometrium. In immunohistochemical detection, immunoreactivity for P450scc was detected in most trophoblast cells, but not in nonpregnant uteri, suggesting that trophoblast epithelial cells synthesized and secreted the sex steroid hormones and/or their precursors to maintain the pregnancy in the Antarctic minke whale.     

Infertility associated with the absence of endometrial progesterone receptors in a bitch

A three‐year‐old intact female Old English sheepdog was presented for evaluation of infertility. A uterine biopsy was performed during dioestrus, and the microscopic appearance was inconsistent with progesterone stimulation; the glands were sparse, simple and failed to show coiling, while the glandular epithelium was cuboidal instead of columnar. There was very little evidence of glandular activity. Due to the inappropriate appearance of the glands for the stage of the cycle, immunohistochemistry for progesterone receptors was performed. No progesterone receptor‐positive immunoreactivity was identified in the endometrial luminal epithelium, glandular epithelium or stroma. Weak intranuclear immunoreactivity was identified within the smooth muscle cells of the myometrium. The absence of progesterone receptors within the endometrial glands is the most likely explanation for the abnormal appearance of the endometrium and for this bitch's infertility. To our knowledge, this is the first report of endometrial progesterone receptor absence in a bitch.     

Localization of interleukin-6 receptor mRNA in the pregnant and non-pregnant mouse uterus

To understand roles of interleukin 6 (IL-6) family cytokines for pregnancy in mice, localization of IL-6 receptor (IL-6R) mRNA was investigated in non- and early pregnant uteri by in situ hybridization. IL-6R mRNA was expressed in all non-pregnant uteri and in pregnant uteri from the third day (Day 3) to the sixth day of pregnancy (Day 6; the day of plug = Day 1). IL-6R mRNA signals were detected in non-pregnant mice in the luminal and glandular epithelium. Signal strength varied according to the sexual cycle. There was no correlation between the signal strength of the IL-6R mRNA and the serum concentrations of progesterone and 17beta-estradiol, which show a monophasic rise in the non-pregnant sexual cycle. In pregnant mice, slight signals were detectable in the luminal and glandular epithelium on Day 3. IL-6R mRNA messages increased with progression towards Day 4, however, localization changed drastically on Day 5. Stromal cells abruptly expressed their mRNA on Day 5, and these cells strongly expressed it on Day 6. The function of IL-6R in the luminal and glandular epithelium might be different from that in the stroma during the implantation period. In addition, few signals were identified in the stromal cells adjacent to the luminal epithelium on Day 6. This suggests that there are two types of stromal cells on Day 6 in mice.     

A comparative study of ovine placental lactogen and prolactin secretion patterns in genetically dissimilar fetal co-twins

Simultaneous fetal and maternal ovine placental lactogen (oPL) and prolactin (oPRL) measurements were determined from blood samples collected from six mixed breed (MB; gestation = 141.5 +/- .7 d) pregnancies and single Rambouillet (n = 2; Ra gestation = 148 and 149 d) and Finnish Landrace (n = 2; Finn gestation = 141 and 143 d) pregnancies for the purpose of examining a possible relationship between fetal-maternal oPL and oPRL levels in the prepartal period. The MB pregnancies were produced by embryo transfer and consisted of genetically different fetuses co-existing in a common uterine environment. Despite the apparent prematurity of Ra co-twins in MB pregnancies, similar oPL and oPRL patterns during the final 14 d of gestation were observed for both Ra and Finn co-twins. Fetal oPL levels decreased at parturition (range 23 to 90 ng/ml) to approximately 60% of values recorded 8 d prepartum (range 100 to 150 ng/ml) in both fetal siblings. Increases in fetal oPRL concentrations, first observed at approximately 4 d prepartum in all fetuses, reached peak concentrations (40 to 60 ng/ml) at term. Results of this study suggest a similarity in regulation of fetal oPL and oPRL secretion, despite genetic differences for gestation period, in fetuses in the M.B. pregnancy.     

妊娠期牦牛肉阜间区子宫腺的形态计量学分析

运用形态计量学方法对31头牦牛肉阜间区子宫腺的相关组织学数据进行了测定,并进行统计分析。研究发现,除了妊娠151~180 d,牦牛肉阜间区子宫腺上皮高度随着妊娠时间的增长极显著降低(P0.01);妊娠期子宫腺管周长和面积非常明显增大(P0.01);不同妊娠阶段之间体积密度差异极显著(P0.01),先增加后下降,而表面密度无显著差异(P0.05);除了妊娠151~180 d,扭曲腺管和管状腺管的比例随着妊娠时间的增长分别非常显著地增大或减小(P0.01);分泌腺管的比例在妊娠期间有明显变化(P0.05),牦牛肉阜间区还存在嵌套腺管。结果显示,妊娠期牦牛肉阜间区子宫腺形态结构有明显的变化,子宫腺变的更大、更长,形状更加扭曲。     

Expression of interleukin-18 receptor mRNA in the mouse endometrium

Interleukin-18 (IL-18) is a proinflammatory cytokine involved in chronic inflammation, autoimmune diseases, and a variety of cancers, and is expressed in mouse uteri. Our previous study suggested that IL-18 acts as a paracrine factor, regulating endometrial function. To elucidate the physiological roles of IL-18 in the mouse endometrium, the expression of the IL-18 receptor (IL-18R) alpha subunit was analyzed. IL-18Ralpha mRNA was expressed in several mouse organs in addition to the endometrium. In situ hybridization analysis using a biotin-labeled mouse IL-18Ralpha riboprobe demonstrated that IL-18Ralpha mRNA expression was detected in glandular epithelial cells, stromal cells around uterine glands, and myometrial cells in the mouse uterus, suggesting that these cells are targets for IL-18. The uterine IL-18Ralpha mRNA expression level changed with the estrous cycle. The uterine IL-18Ralpha mRNA levels of estrous mice were higher than those of diestrous mice. In addition, the IL-18Ralpha mRNA levels in uteri at 3 and 14 days after ovariectomy were higher than those at diestrus and decreased following treatment with estradiol-17beta or progesterone. These findings suggest that IL-18Ralpha gene expression is regulated by estrogen and progesterone and that the uterine IL-18 system is involved in the regulation of uterine functions in a paracrine manner.     

Steroid receptor expression and HER-2/neu (c-erbB-2) oncoprotein in the uterus of cats with cystic endometrial hyperplasia-pyometra complex

The presence of oestrogen-alpha receptor (ER), progesterone receptor (PR), and HER-2/neu (c-erbB-2) oncoprotein in the uterine walls of 10 healthy cats and 20 subjects with cystic endometrial hyperplasia-pyometra (CEH-P) were evaluated. Lesions were graded according to the severity of cystic dilation, hyperplasia and inflammation, and were classified as normal, mild uterine hyperplasia and severe uterine hyperplasia. The ER, PR and c-erbB-2 expression in the endometrium, glandular epithelium, stromal fibroblasts and myometrial smooth muscle cells was quantified by immunohistochemistry. The ER, PR and c-erbB-2 staining patterns differed between normal uteri and uteri with CEH-P. The ER expression was tended to be higher in the endometrial surface and glandular epithelium in the severe hyperplasia group (P > 0.05) and significantly lower in the mild hyperplasia cases compared with normal endometrium (P < 0.05), whereas the PR expression in both severe and mild hyperplasia cases tended to be higher in stromal cells and glandular epithelium than those in the normal uteri. C-erbB-2 immunoreactivity was observed only in the endometrial surface and glandular epithelium of the uterine wall and immunostaining was found to be highest in cases with severe hyperplasia. As a conclusion, we suggest that c-erbB-2 oncoprotein may play a role in the pathogenesis of the CEH together with the ER and PR in cats, and that ER does not have a role in the mechanism of pyometra, whereas PR plays a role in the pathogenesis of both CEH and pyometra.     

Retinol and estradiol regulation of retinol binding protein and prostaglandin production by porcine uterine epithelial cells in vitro

Secretion into the uterine lumen follows a precise pattern during early pregnancy. Near the end of the second week of pregnancy and coincident with elongation of conceptuses, retinol, retinol binding protein (RBP), estradiol (E2), and prostaglandins E (PGE) and F (PGF) increase in the uterine lumen, and RBP mRNA increases in the endometrium. In the present studies the potential for E2 (0.1 microM) and retinol (10 microM) to regulate RBP and PG production by cultured luminal (LEC) and glandular (GEC) epithelial cells collected from postpubertal females and LEC from prepubertal gilts was examined. Endometrial tissue was collected surgically from cyclic and pregnant females (n = 8) on d 10 and 13 postestrus (first day of estrus = d 0) and from 120- and 150-d-old prepubertal gilts that were treated with progesterone (P4) (2.2 mg x kg(-1) x d(-1), n = 6) or corn oil (n = 6) for 14 d prior to tissue collection. The LEC from postpubertal females responded to retinol with increased (P < 0.05) RBP, PGE, and PGF in culture medium and increased (P < 0.07) RBP mRNA but E2 decreased (P < 0.05) RBP and RBP mRNA and had no effect on prostaglandins. No E2 or retinol effects on secretions of GEC occurred in vitro, but a day x pregnancy status interaction (P < 0.06) affected PGE output by the GEC. Secretion of PGE was greater when GEC were collected on d 10 of pregnancy than from d-10 cyclic or d-13 pregnant or cyclic females. Both E2 and retinol stimulated (P < 0.05) secretion of RBP by LEC isolated from prepubertal gilts, but their effects were not additive. In vivo treatment of prepubertal gilts with P4 increased (P < 0.05) RBP and decreased (P < 0.05) PG production by LEC in vitro. Therefore responses to E2 and retinol differ between pre- and post-pubertal females, and retinol may function in the regulation of endometrial RBP and PG secretion.     

Uteroferrin (ACP5) in the Ovine Uterus:Ⅰ. Regulation by Pregnancy and Progesterone

Uteroferrin,also known as type 5 tartrate resistant acid phosphatase ( ACP5 ) or TRAP,is an iron-containing glycoprotein secreted by uterine gland epithelium (GE) in response to progesterone and transported across the placental areoalae into the fetal circulation and allantoic fluid to deliver iron and to stimulate hematopoeisis in pigs.This study determined if ACP5 was expressed in the ovine uterus in response to pregnancy,progesterone,interferon tau,placental lactogen,and placental growth hormone.ACP5 protein was present in uterine GE of cycfic and early pregnant ewes,particularly between days 18 and 120 of pregnancy.ACP5 mRNA was expressed in uterine GE of cyclic and pregnant ewes in the same temporal and cell-specific manner.ACP5 was present in secretions from uterine glands,i.e.,uterine milk,and aUantoic fluid from days 40 to 80 of pregnancy,and in uterine flushings from cyclic and early pregnant ewes.Progesterone induced expression of ACP5 mRNA and intrauterine infusion of recombinant ovine interferon tau further stimulated ACP5 expression in uterine GE of ewes,but intrauterine injections of ovine placental lactogen and ovine growth hormone had no effect on ACP5 expression in uterine GE.These results indicate that ACP5 is:1 ) expressed only in GE in response to progesterone ;2 ) secreted into the uterine lumen and transported into the conceptus via plaeental areolae during pregnancy;and 3) present in secretions from uterine GE and in allantoic fluid.The roles of ACP5 in the ovine uterus may include transport of iron across the placenta and stimulation of hematopoiesis.     

The impacts of uterine environment and fetal genotype on conceptus size and placental vascularity during late gestation in pigs.

Meishan and Yorkshire gilts were bred exclusively to Yorkshire and Meishan boars, respectively, resulting in similar Meishan x Yorkshire fetuses gestating in the uteri of both maternal breeds. Gilts of both breeds were slaughtered on d 90 and 110 of gestation, and the weight and volume of each uterine horn was determined. Fetal weights and crown-rump lengths were recorded. A section of the chorioallantoic-endometrial attachment site was collected for each conceptus and evaluated for placental and endometrial vascular density. An intact placenta was recovered for each conceptus and weighed, and its surface area was determined. Fetal weight and crown-rump length increased (P < .001) markedly between d 90 and 110 of gestation in Meishan and in Yorkshire uteri, but they were markedly less (P < .001) in Meishan than in Yorkshire uteri. Placental weight and placental surface area were reduced by 40% in Meishan, compared with Yorkshire, uteri; however, placental size did not increase between d 90 and 110 in either uterine type. Placental vascular density and associated endometrial vascular density were similar (P > .20) for conceptuses in Meishan and Yorkshire uteri on d 90 and 110 of gestation. Additionally, even though the ratio of fetal weight: placental weight (placental efficiency) increased between d 90 and 110, placental efficiency was similar for conceptuses in Meishan and Yorkshire uteri. In a previous study using straightbred Meishan or Yorkshire conceptuses gestated in either a Meishan or Yorkshire uterus, we found Meishan conceptuses had a markedly greater placental efficiency than did Yorkshire conceptuses, regardless of the type of uterus in which they were gestated. These data indicate that uterine type determines conceptus size and conceptus genotype controls placental efficiency.