Apoptosis in the porcine uterine endometrium during the estrous cycle, early pregnancy and post partum

The mammalian uterus changes dramatically during the estrous cycle, pregnancy, and involution post partum. Dynamic changes in the uterine endometrium are a type of homeostasis and proceed with proliferation and exclusion of cells. Homeostasis of the uterus is closely related to apoptosis involving various hormones and cytokines. The objective of the present study was to determine the morphological features and occurrence of apoptosis in the porcine endometrium during the estrous cycle, early pregnancy, and post partum. Cyclic changes in the morphology of the surface epithelium were observed during the estrous cycle. The heights of surface epithelia were significantly high on day 4 of the estrous cycle and the early pregnancy. The heights of the surface epithelium remained low from days 1 to 31 post partum. We then used terminal deoxynucleotidyl transferase-mediated biotinylated deoxyuridine triphosphate nick end-labeling (TUNEL) of the 3'-terminal of fragmented DNA, which is effective for detection of apoptosis in various tissues. We found that apoptosis in the porcine endometrium contributed to homeostasis of the endometrium during the estrous cycle through control of cell proliferation and exclusion. Conversely, apoptosis on days 4 and 8 of gestation before the implantation window depended on the plasma estrogen and progesterone levels; however, suppressive homeostasis of apoptosis occurred at the time of implantation on days 15, 18 and 21 of gestation. Our study is the first to demonstrate apoptotic cell death in the porcine endometrium directly by TUNEL method. The results strongly suggest that uterine homeostasis is mainly controlled by apoptosis during the estrous cycle and early pregnancy.     

secretion of the major progesterone-induced proteins of the sheep uterus by caruncular and intercaruncular endometrium of the pregnant ewe from days 20–140 of gestation

An experiment was performed to evaluate the types and quantity of proteins secreted by intercaruncular endometrium at days 20, 60, 100 and 140 of gestation and caruncular endometrium from days 100 and 140 of gestation. Tissues were obtained from ewes made unilaterally pregnant. Based on SDS polyacrylamide gel electrophoresis (SDS-PAGE), the major proteins present in uterine fluid at days 60–140 of gestation were the uterine milk proteins (UTM-proteins), a pair of structurally-related, progesterone-induced polypeptides with molecular weights of 55,000 and 57,000. These proteins were also present in uterine fluid at day 20, but the major protein at this time migrated coincident with albumin. Cultured explants of endometrium at all days of pregnancy produced UTM-proteins as their major radiolabelled product for both caruncular and intercaruncular endometrium. The amount of protein secretion in vitro was greater (P<.04) for intercaruncular endometrium than for caruncular endometrium but was not significantly affected by stage of gestation or local presence of the conceptus. Immunohistochemical analysis revealed that UTM-proteins were present in glandular and luminal epithelium of intercaruncular endometrium and in both epithelial and stromal elements of caruncular endometrium. It was concluded that the UTM-proteins are produced earlier than previously described (i.e., day 20). In addition, caruncles contribute to the uterine secretory protein milieu through the secretion of proteins that are similar to that produced by the glandular intercaruncular epithelium.     

产后奶牛子宫内膜的扫描电镜观察

应用扫描电镜观察了 5头奶牛产后不同时期子宫内膜的变化。结果发现 ,分娩后早期 ,奶牛子宫内膜部分破损 ,与破损部位相邻的细胞在产后 1周内相继脱落 ;未破损内膜上皮细胞表面的纤毛和微绒毛形状不整、数量减少。产后2周 ,基底细胞开始生长 ,至产后 4 5 d时生长完成 ;产后 4周 ,内膜破损区域缩小 ,上皮细胞表面纤毛和微绒毛形状规则、数量增加 ;产后 4 5 d,偶见上皮细胞缺失 ;至产后 6 0 d,子宫内膜上皮细胞完整、排列有序、被覆纤毛和微绒毛。上述结果表明 ,奶牛子宫内膜在分娩后 6 0 d内完成修复。     

Distribution of androgen receptor in the porcine uterus throughout pregnancy.

The uterus is a well-known target of endocrine, paracrine and autocrine acting molecules among which steroid hormones (oestrogens, androgens and progesterone) are of special importance. The uterine tissues (endometrium and myometrium) undergo morphological and physiological changes which are associated with changes in expression of steroid hormone receptors. Androgen receptors (AR) that mediate the action of androgens have already been detected in porcine uteri during the oestrous cycle and early pregnancy. To evaluate the role of AR in uterine physiology, the presence of ARmRNA and AR protein localization in the porcine uterus from day 10 to day 90 of pregnancy and in the uterus from the foetus of day 90 postcoitum (p.c.) and from the neonatal 1-day-old piglet was studied. ARmRNA was detected in the porcine endometrium up to day 18 p.c., while AR protein was detectable in glandular epithelium and stromal cells as through day 90 of pregnancy. AR was also detected in the myometrium on all investigated days of pregnancy; however, on day 90, the immunostaining was present only in a limited number of cells. AR immunostaining was clearly demonstrated in the uterus of the female foetuses on day 90 as well as in the uterus of 1-day-old piglets. The physiological relevance of this finding needs further elucidation.     

Distribution patterns of mast cells in the uterus of pregnant Meishan pigs

The present study was performed to investigate the numerical distribution of mast cells (MCs) in the uteri of pregnant Meishan pigs to explore the functions of MCs in pig pregnancy. The uterine samples from pregnant (on days 15, 26 and 50 of gestation) pigs were obtained respectively and stained with toluidine blue. The results were as follows: MCs were constitutively located in the uterus of the Meishan pig, with the distribution varying with gestational stages. On days 15 and 26 of gestation, MCs were mainly distributed around the blood vessels and uterine glands within the endometrium. On day 50 of gestation, MCs were mostly distributed in the myometrium. These results indicated that uterine MCs possibly have versatile functions in pig pregnancy.     

Expression of amphiregulin during the pre- and post-implantation period in the mouse reproductive tract

In mammals, embryo implantation is an essential step in reproduction. Implantation is a phenomenon that involves crosstalk between the blastocyst and the maternal endometrium. However, the molecular basis of the connections between the blastocyst and endometrium is not yet fully understood. Amphiregulin is a member of the epidermal growth factor family and is known to be expressed in the luminal epithelium of the mouse uterus on 3.5 days post coitum (dpc). Thus, to clarify the mechanism of amphiregulin at fetomaternal interface, we analyzed the expression pattern of amphiregulin mRNA in the oviducts and uteri of pregnant and psuedopregnant mice by means of real-time PCR. Amphiregulin expression in the pregnant uterus dramatically increased on 2.5 dpc, peaked on 3.5 dpc, and declined by 5.5 dpc. Furthermore, to analyze the effect of the presence of an embryo on amphiregulin expression, we determined the expression pattern of amphiregulin mRNA in the uterus after embryo transfer on 0.5 and 1.5 dpc. A previous study showed that the expression of amphiregulin mRNA depends on the concentration of progesterone. However, our present results indicate that amphiregulin mRNA is upregulated by the presence of fertilized eggs in the lumen of the oviduct on 0.5 dpc.     

八点黑獭兔不同发育时期子宫组织结构变化的研究

观察母兔妊娠20、26、29天和15、30、60、90日龄八点黑獭兔的子宫组织,以探讨八点黑獭兔子宫的形态学发育特点。结果表明：獭兔的子宫随着年龄的增长．其内膜的粘膜上皮由假复层柱状纤毛上皮逐渐分化成紧密规则的单层柱状上皮;固有层在15～30日龄之间出现．内含大量间质细胞;妊娠期固有层发生水肿和充血;子宫肌层在幼龄时分层不明显,各层肌纤维互相交织,随平滑肌细胞分裂增生,3月龄时肌层出现明显的内环、外纵两层,中间形成一层环状排列的血管：獭兔出生后其子宫内膜腺由子宫内膜上皮细胞内陷形成,随后腺上皮细胞经有丝分裂增生,15日龄幼兔已形成单管子宫腺。该研究结果可为兔的解剖学、生殖生理学增添新的内容,也为进一步研究兔生殖系统疾病的治疗与预防提供参考资料。     

Expression and Hormonal Regulation of IL-11Rα in Canine Uterus During Early Pregnancy

Embryo implantation is critical for the successful establishment of pregnancy. Interleukin-11 (IL-11) is essential for adequate decidualization in the mouse and human via binding to the specific IL-11 receptor α (IL-11Rα). But the expression and regulation of IL-11 and IL-11Rα in the canine endometrium remain unknown. The aim of this study was to investigate the differential expression of IL-11Rα in canine uterus during early pregnancy and its regulation under different conditions by in situ hybridization. Interleukin-11Rα mRNA was mainly localized in glandular epithelium in canine uterus. There was a low level of IL-11Rα expression in the glandular epithelium on days 6, 12 and 17 of pregnancy. On day 20 of pregnancy when embryo implanted, IL-11Rα mRNA was highly expressed in the glandular epithelium surrounding the embryo, but not in the luminal epithelium and stroma. On day 23 of pregnancy, the expression of IL-11Rα mRNA maintained a constant level compared with the expression of day 20 and increased on day 28 of pregnancy. During the oestrous cycle, a high level of IL-11Rα mRNA expression was seen in the oestrous uterus. Progesterone slightly induced the expression of IL-11Rα mRNA in the ovariectomized canine uterus. These results suggest that IL-11Rα expression is closely related to canine implantation and up-regulated by progesterone.     

The dynamic expression of extracellular matrix in the bovine endometrium at implantation

Remodeling of uterine endometrial extracellular matrix (ECM) is pivotal to successful implantation and placentation, and has been well described in the rodents and humans. However, bovine endometrial ECM remodeling is still vaguely defined, especially at the time of implantation. Therefore, this study investigated the distribution of four ECMs namely, types I and IV collagen, laminin and fibronectin, from days 0 to 30 of gestation in bovine endometrium by immunofluorescence microscopy. A change in the distribution pattern of ECMs was evident by day 14 of gestation as features at this stage were clearly different from those of day 14 of the estrous cycle. The immunoreactivity of type I collagen, fibronectin and laminin decreased from day 14 of gestation and was obscured by day 24 of gestation. The type I collagen fibers formed were of thinner consistency than those of the estrous cycle and showed a coarser meshwork within the epithelium sites during the implantation period. In addition, the type IV collagen and laminin immunoreactivities of epithelial basement membrane also remarkably declined at exactly the same time. By day 30 of gestation, the four ECMs had regenerated with the formation of the placentome. In conclusion, this study reveals that remodeling of ECM is essential for the successful establishment of pregnancy in the bovine.     

Expression of Oestrogen Receptor α and Oestrogen Receptor β in the Uterus of the Pregnant Swine

The uterus is a well‐known target of endocrine, paracrine and autocrine acting molecules among which steroid hormones are of special importance. The objective of our work was to localize oestrogen receptors (ERα and ERβ) mRNA and protein in the pig uterus throughout pregnancy (10, 18, 32, 50, 71, 90 days post coitum) using RT‐PCR, Western‐blot and immunohistochemistry. The present study is the first one to demonstrate the presence of ERs protein in the porcine uterus not only at the beginning but also at mid‐ and late pregnancy. In the pregnant swine, ERα was immunolocalized in the luminal epithelium (LE) and glandular epithelium (GE) and the myometrium of the uterus with differences in the intensity of staining at different stages of pregnancy studied. The LE and GE of pregnant swine stained for ERβ regardless of the day of pregnancy examined, whereas only a few cells within the myometrium showed a weak immunoreactivity. Western blot analysis confirmed the presence of ERα and ERβ proteins on all investigated days of gestation. The expression of ERα and ERβ mRNA was detected by RT‐PCR in all examined samples corresponding to each of the consecutive stages of pregnancy. The obtained results show that ERα is more abundant in comparison to ERβ within the porcine pregnant uterus. The presence of ERα and ERβ in all compartments of the pig uterus during pregnancy may indicate direct action of oestrogens on proliferation and differentiation of these cells.     

小鼠子宫内膜凝集素结合特性的生殖周期改变

用４种酶标植物凝集素（ＣｏｎＡ,ＷＧＡ,ＳＢＡ,ＲＣＡ）作探针,研究了小鼠发情周期、妊娠期和产后子宫内膜上皮、固有膜细胞和腺上皮凝集素结合特性的变化。子宫内膜与这４种凝集素的结合,随发情周期而波动。与间情期相比,妊娠期子宫内膜上皮与ＣｏｎＡ的结合,由阴性转为阳性,并随妊娠的持续逐渐加强,产后又突然转为阴性。子宫内膜上皮与ＷＧＡ的结合,在间情期和妊娠１ｄ均为阳性,妊娠７ｄ减为弱阳性,继之又加强,产后达最强。无论妊娠期还是产后,子宫内膜上皮与ＲＣＡ的结合均为强阳性,与ＳＢＡ的结合均为阴性。结果提示,生殖周期子宫内膜与凝集素结合特性的规律性变化,可能与激素调节、胚泡的着床和分娩有关     

Interleukin-1 receptor antagonist expression in the equine endometrium during the peri-implantation period

To identify factors involved in the establishment of pregnancy in the mare, endometrium was collected from day 13 (day 0 = day of ovulation) cyclic and day 13, 19, and 25 pregnant animals. From initial cDNA subtraction studies, interleukin-1 receptor antagonist (IL-1RN) mRNA was found as a candidate molecule expressed uniquely in the pregnant endometrium. Expression of IL-1RN mRNA was markedly increased in day 19 and 25 gravid endometrium. In situ hybridization analysis revealed that IL-1RN mRNA was localized to the glandular epithelium. Interleukin-1 receptor antagonist (IL-1RN) protein was found in the extracts of day 25 gravid endometrium and was immunochemically localized to the glandular epithelium/luminal cavity of the pregnant uterus. High concentrations of estradiol-17β (E₂) were detected in day 25 conceptuses. Concentrations of E₂ were higher in the gravid endometrial portion than in other endometrial regions. On the other hand, progesterone concentrations did not differ among endometrial samples analyzed. Furthermore, the expression of IL-1RN mRNA was up-regulated in endometrium culture samples treated with 10 ng/mL E₂ and 10 ng/mL progesterone. In the analysis of related gene expression, increased amounts of IL-1α and IL-6 mRNA were also found in the day 25 gravid endometrium; however, these expressions in endometrial culture samples were not up-regulated by the steroid treatment. These results indicate that expression of IL-1RN in the endometrium is likely regulated by E₂ and progesterone and suggest that IL-1RN regulates the degree of IL-1 signal transduction and thereby plays an important role in the establishment of equine pregnancy.     

Tumour necrosis factor in the canine endometrium: an immunohistochemical study

Tumour necrosis factor (TNF), a pleiotropic cytokine that regulates cell growth and differentiation as well as the synthesis of other cytokines, has been identified in the uterus of several species describing a cyclic pattern, eventually under ovarian steroid regulation. Information is yet limited on the presence of TNF protein in the canine endometrium during the oestrous cycle and early pregnancy. This study depicts the temporal immunolocalization of TNF in the bitch endometrium along the oestrous cycle and changes associated with the early steps of embryo invasion. TNF immunolabelling was found in both the stromal fibroblasts and epithelial components of the canine endometrium in all stages studied. Stromal immunostaining was more intense than that of the epithelia, in all the stages of the oestrous cycle. In addition, a tendency for a decrease in the surface epithelium intensity score was found in early dioestrus. A positive glandular content was only observed in anoestrus and proestrus stages. In early pregnancy (days 13-16), TNF immunolabelling was detected at the embryo-maternal surface, in the syncytium cords and the trophoblast, as well in the endometrial stroma and the basal endometrial glands, but not in the lacunar epithelium. The overall TNF immunoreactivity was higher in early pregnancy samples in comparison with those of the early dioestrus and dioestrus stages, suggesting it plays a role during implantation.     

Uterine secretions of the cow contain proteins that are immunochemically related to the major progesterone-induced proteins of the sheep uterus

Proteins that cross-react with antiserum to the major progesterone-induced proteins found in the pregnant sheep uterus, the uterine milk proteins (UTM-proteins), were detected as radiolabelled secretory products of endometrium from pregnant cows. Cross-reactive proteins included a form at 57,000 molecular weight as well as other lower-molecular-weight variants found in lower amounts. Similar proteins were also detected in uterine fluid from a cow at day 270 of gestation. Using immunohistochemical procedures, proteins that cross-reacted with antiserum to sheep UTM-proteins could be localized to the epithelial cells of endometrial glands in the cow. Results indicate that UTM-protein-like molecules are secreted by the endometrium of the cow during pregnancy.     

Gastrointestinal Diseases of Marsupials

This article focuses on gastrointestinal diseases that affect 3 of the most common marsupials treated by veterinarians: the red kangaroo (Macropus rufus), Bennett’s wallaby (Macropus rufogriseus), and the sugar glider (Petaurus breviceps). Other marsupial species maintained in captivity include the eastern gray kangaroo (Macropus gigantus), the tammar wallaby (Macropus eugenii), and the wallaroo (Macropus robustus). Most all of the disease conditions discussed in this article are also diagnosed in the lesser species listed earlier. Marsupials derive their name from the presence of the marsupial bones (ossa marsupialia), which serve as the surface for the attachment of several abdominal muscles. The ossa marsupialia were once thought to support the marsupium (pouch), although this was an incorrect assumption. The metabolic rate of marsupials is generally considered to be approximately two-thirds that of eutherian or placental mammals. Marsupials belong to a unique group of animals whose development is characterized by a very short gestation, the birth of relatively underdeveloped young (joeys), and a lengthy period of lactational development that typically occurs in a pouch. The physiologic differences that exist between marsupials and eutherian mammals should be considered when managing disease conditions in these animals.     

Expression profiles of perforin,granzyme B and granulysin genes during the estrous cycle and gestation in the bovine endometrium

The conceptus is susceptible to destruction by maternal cytotoxic lymphocytes, which have cytotoxic potential. Therefore, it is expected that mechanisms for regulating cytotoxic lymphocytes exist, but little is known about the expression of cytotoxic genes in the endometrium. In the present study, we examined the spatial and temporal expression patterns of the cytotoxic genes perforin, granzyme B, and granulysin during the estrous cycle and gestation in the bovine endometrium. Endometrial tissues were collected from cows during the estrous cycle and gestation. The gene expression patterns of the three cytotoxic genes were examined using quantitative polymerase chain reaction and in situ hybridization, and cytotoxic lymphocyte subsets were characterized using immunohistochemistry. During mid‐ to late gestation in the intercaruncular (ICAR), granulysin expression was significantly increased, and a large number of granulysin‐expressing cells were localized in the luminal epithelium. Perforin and granzyme B displayed similar expression profiles and were highly expressed in the peri‐implantation endometrium, but few cells expressing these genes were found in the endometrial stroma. In conclusion, these findings suggest that in the ICAR epithelium granulysin may play important roles in the establishment and maintenance of gestation during normal pregnancy.     

Exogenous somatotropin alters IGF axis in porcine endometrium and placenta

The aim of this study was to examine whether exogenous somatotropin (ST) can alter the insulin-like growth factor (IGF) axis in the porcine epitheliochorial placenta. Crossbred gilts were injected either 6 mg of recombinant porcine ST or vehicle from days 10 to 27 after artificial insemination (term day 116). Control and ST-treated gilts were euthanized on day 28 (8 control/5 treated), day 37 (4 control/6 treated), and day 62 (4 control/6 treated) of gestation. Endometrium and placental tissue samples were collected and subjected to mRNA analyses. In control gilts, somatotropin receptor (STR) and IGF-I mRNA abundance in the endometrium decreased with gestation. Conversely, the amounts of IGF-II mRNA and of IGF binding protein (BP)-2 and -3 mRNA, which were analyzed in endometrium and placental chorion, increased with gestation. The endometrium contained less IGF-II mRNA but more IGFBP-2 and-3 mRNA than the placental chorion. In response to pST treatment, the amounts of endometrial STR and IGF-I mRNA were lower at days 28 and 37, but higher at day 62 of gestation. The content of IGF-II mRNA was higher in the endometrium of pST-treated than control gilts on day 37. The amount of IGFBP-2 mRNA was increased on day 37 in endometrium and placenta of pST-treated gilts, whereas no changes in IGFBP-3 mRNA were observed. The IGF-II/IGFBP-2 ratio was higher in the placenta in response to pST on day 28 of gestation. Results show that pST treatment of pregnant gilts during early gestation alters IGF axis in maternal and fetal placental tissues and suggest pST may exert an effect on fetal growth by altering the relative amount of IGFBPs and IGFs at the fetal-maternal interface.     

The uterus and early pregnancy failure in the mare

The environment the equine conceptus finds itself in when it arrives in the uterus some 6 days after ovulation will determine if it will thrive or die. The uterus will be its home for approximately the next 11 months and as such it needs to both coordinate the growth of the placenta and ensure there is enough nourishment passing to this organ for the fetus to develop normally. The trophoblast, endometrium, maternal ovaries and, later in pregnancy, the fetal gonads, all play roles in the hormonal changes that orchestrate these events. Although failures of these processes later in pregnancy can have catastrophic effects for the fetus, it is in early gestation that the foundations for a successful pregnancy are laid. This paper therefore concentrates on some of the noninfectious influences the uterus may have on survival of the young conceptus.