Blastocyst implantation in the domestic cat. Light and electron microscopic study]

Blastocyst implantation in the domestic cat A light and electronmicroscopic investigation Implantation in the cat begins 121/2 days post coitum and lasts to the 14th day. Several hours before implantation begins the extraembryonic trophoblast, in what could be considered a precontact or sensitization phase, shows fine structural evidence of secretory activity. The uterine epithelium at the same times gives evidence of being at first resorptive and, after a differentiation, of being secretory. Secretion also in the gland epithelium can be observed. The attachment of the embryo to the endometrium begins with an appositional phase. The trophoblast is now distinctly resorptive and forms with the uterine epithelium microvilli-free areas of contact lying between the openings of the uterine glands. Then follows an adhesion phase during which additional junctional complexes are formed between an actively proliferating trophoblast and the uterine epithelium which shows now symplasmic differentiation. Finally, in the intrusion phase the uterine epithelium completely disintegrates. In this manner the syncytiotrophoblast which differentiates from the cytotrophoblast invades the maternal tissues until it reaches the endothelium of the uterine blood vessels.     

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.     

Unusual phenomena during pregnancy in the cat and cow.

Unusual situations were observed during examination of pregnant uteri from a cat and a cow. The feline specimen revealed three corpora lutea at the surface of each ovary with three fetuses developing normally in the left and two in the right uterine horn. The right horn had been ruptured between two of the locular enlargements but the damage was sealed by the omentum. Three conceptuses were present in the cow's uterus. Two of these embryos, each with its individual allantois, were developing within a single chorion within one uterine horn. The adjacent tips of these allantoic vesicles were entangled into a complex knot which could provide opportunity for vascular anastomosis.     

Ultrastructural Studies of Chicken Embryo Chorioallantoic Membrane during Incubation

Structural changes taking place in the chorioallantoic membrane (CAM) of embryonated hen's eggs during incubation were followed up by histological, histochemical and electron microscopic methods. The allantoic sac surrounds the amnionic sac and also the yolk sac by the 7th to 11th day, respectively, and forms together with the chorion the CAM. The mesenchymal layer placing between the chorion of ectodermal and the allantoic layer of entodermal origin develops from the somatic and splanchnic-pleure of the parietal mesoderm. The CAM is composed of three different layers, i.e. chorionic epithelium, mesenchyme and allantoic epithelium. The chorion comprises 2 layers of cubical epithelial cells. In between the 2 layers capillaries and directly under the shell membrane vascular sinuses can be found. Blood circulating in the sinuses is separated from the air in the pores of the shell membrane by the adacent epithelial cell-projections of 0,2 μm thickness, the basal membrane (0.1 μm) and the sinus endothelial layer of 0.2 μm thickness. The mesenchymal cells are of star-like form. The allantioc epithelium is built of one layer of fusiform cells with oval-shaped or rod-like nuclei.     

猪子宫腔液外泌体来源的TIMP2蛋白对胚胎附植的影响

旨在研究猪子宫腔液外泌体来源的TIMP2蛋白对妊娠早期胚胎附植的影响.本研究采用超速离心法分离妊娠第9天(n=3)和第12天(n=3)大白猪子宫腔液外泌体,利用Western blot(WB)检测外泌体中TIMP2蛋白的变化,并通过免疫组化试验研究TIMP2蛋白在子宫中的表达定位.进一步用外泌体与猪胚胎滋养层细胞(po...     

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.     

Structure and functions of the placenta in common minke (Balaenoptera acutorostrata), Bryde’s (B. brydei) and sei (B. borealis) whales

The structure and functions of placentas were examined in 3 species of rorqual whales, common minke (Balaenoptera acutorostrata), Bryde’s (B. brydei) and sei (B. borealis) whales, with the aim of confirming the structural characteristics of the chorion, including the presence of the areolar part, and clarifying steroidogenic activities and fetomaternal interactions in the placentas of these whales. Placentas were collected from the second phase of the Japanese Whale Research Program under Special Permit in the North Pacific (JARPN II). Histological and ultrastructural examinations revealed that these whale placentas were epitheliochorial placentas with the interdigitation of chorionic villi lined by monolayer uninucleate cells (trophoblast cells) and endometrial crypts as well as folded placentation by fold-like chorionic villi. Moreover, well-developed pouch-like areolae were observed in the placentas, and active absorption was suggested in the chorionic epithelial cells of the areolar part (areolar trophoblast cells). Berlin blue staining showed the presence of ferric ions (Fe³⁺) in the uterine glandular epithelial cells and within the stroma of chorionic villi in the areolar part. An immunohistochemical examination revealed tartrate-resistant acid phosphatase (TRAP; known as uteroferrin in uteri) in the cytoplasm of glandular cells and areolar trophoblast cells. This result suggested that, in cetaceans, uteroferrin is used to supply iron to the fetus. Furthermore, immunoreactivity for P450scc and P450arom was detected in trophoblast cells, but not in areolar trophoblast cells, suggesting that trophoblast cells synthesize estrogen in whale placentas. Therefore, we herein immunohistochemically revealed the localization of aromatase and uteroferrin in cetacean placentas during pregnancy for the first time.     

Areolae of the Placenta in the Antarctic Minke Whale (Balaenoptera bonaerensis)

In this study, we examined the existence and structure of areolae and the steroidogenesis of areolar trophoblast cells in the Antarctic minke whale placenta morphologically and immunohistochemically. Placentas were collected from the 15th, 16th and 18th Japanese Whale Research Program under Special Permit in the Antarctic (JARPA) and 1st JARPA II organized by the Institute of Cetacean Research in Tokyo, Japan. The opening and cavity of fetal areolae formed by taller columnar trophoblast cells (areolar trophoblast cells) with long microvilli and a bright cytoplasm, as compared with the trophoblast cells of the chorionic villi interdigitating with the endometrial crypts, were recognized in observations of serial sections. The opening of the areolar cavity was hidden by chorionic villi with areolar trophoblast cells. Furthermore, a closed pouch-like structure lined by tall columnar cells similar to areolar trophoblast cells within the stroma of chorionic villi was noticed and continued to the areolar cavity, with the opening seen on serial sections. In a surface investigation of the chorion and endometrium by SEM, maternal (endometrial) areolae irregularly surrounded by endometrial folds were obvious. Moreover, we distinguished areolar trophoblast cells with long microvilli attached with many blebs from trophoblast cells. In our immunohistochemical observations, a steroidogenic enzyme, cytochrome P450 side chain cleavage enzyme (P450scc), was detected with strong immunoreactivity in trophoblast cells. However, areolar trophoblast cells showed weak or no immunoreactivity for P450scc.     

绵羊妊娠识别、建立和维持的信号系统

妊娠的建立和维持归因于胚体的信号和黄体对孕酮的产生,在反刍动物,胚体滋养层分泌的激素抑制子宫内膜产生前列腺素PGF2α。在发情周期的绵羊,孕酮下调子宫内膜腔上皮和子宫腺上皮孕酮受体基因的表达,并伴随着上皮雌激素受体和催产素受体的增加。在催产素的作用下,子宫开始分泌溶黄体作用的前列腺素F2α。在妊娠绵羊,孕体滋养层产生的干扰素可作用于子宫内膜而直接抑制雌激素受体基因和催产素受体基因的转录。孕酮、干扰素、胎盘催乳素和生殖激素共同组成了一个"作用网络"来调节子宫的功能分化和子宫腺的形态发生,从而维持绵羊妊娠。本文综述了绵羊妊娠识别、建立和维持的信号系统。     

Expression of uterine sensitization-associated gene-1 (USAG-1) in the mouse uterus during the peri-implantation period

Rat uterine sensitization-associated gene-1 (USAG-1) mRNA is expressed in the uterus during the peri-implantation period, and its mRNA expression in uterine epithelial cells is highest on day 5 of pregnancy. On the other hand, since changes in USAG-1 mRNA expression in the mouse uterus are not seen during the estrous cycle, USAG-1 expression might be specifically regulated by embryonic factors rather than by the maternal environment. However, the expression pattern and function of USAG-1 in the mouse uterus have not been determined. Thus, we examined the tissue-specific USAG-1 mRNA expression in the uteri of ICR mice during peri-implantation using real-time quantitative PCR. Uterine tissues, such as the myometrium, luminal epithelium, and stroma, were collected by laser capture microdissection at 3.5-6.5 dpc. USAG-1 mRNA was expressed in the uteri of pregnant mice from 3.5 dpc to 6.5 dpc, and the highest level of expression was seen at 4.5 dpc (P<0.01). Significantly high USAG-1 mRNA expression was detected in the luminal epithelium at 4.5 dpc (P<0.05). The stroma and myometrium exhibited unchanged expression levels of USAG-1 mRNA at 3.5-5.5 dpc. USAG-1 mRNA was undetectable in blastocysts and implanting embryos. Expression of USAG-1 mRNA appears to be associated with blastocyst implantation to the luminal epithelium, suggesting that physiological or biochemical contact of the blastocyst to the uterus is required for USAG-1 expression.     

Cesarean section and ovariohysterectomy after severe uterine torsion in four cows

OBJECTIVE: To report a technique for intra-partum ovariohysterectomy (OVH) in cows after severe uterine compromise after uterine torsion. STUDY DESIGN: Clinical case report. ANIMALS: Four cows with uterine torsion. METHODS: After a ventrolateral cesarean section, uterine closure, and uterine detorsion, the laparotomy incision was elongated (20 cm) caudally so that the uterus can be completely exteriorized. The uterus was separated from broad ligaments by transection between 2 rows of overlapping interrupted sutures placed in the broad ligaments to a point approximately 2 cm caudal to the external os of the cervix. The vagina was occluded with 2 clamps directly caudal to the external os of the cervix and closed with 5 single cruciate sutures that penetrated all layers, and then the vagina was transected between the suture line and external os to remove the uterus. The abdominal incision was closed in layers. Results- All cows survived the surgery; however, 1 died the next day from Escherichia coli sepsis and 1 was euthanatized at 8 days because of unresponsive E. coli mastitis. One cow, 28 days after surgery developed a 15 cm perivaginal abscess, which resolved after drainage. This cow and the fourth cow survived and lactated. CONCLUSIONS: Intra partum OVH is technically possible after cesarean section in cows with uterine compromise or necrosis after uterine torsion. CLINICAL RELEVANCE: Ovariohysterectomy should be considered in cows with severe uterine torsion that have uterine tissue compromise to improve survival.     

Expression Pattern of CD34 at the Maternal–Foetal Interface During Pregnancy in Pigs

The pig exhibits a non‐invasive, epitheliochorial placentation. Adhesion molecules are indispensable for successful implantation and establishment of placentation. CD34 is an adhesion molecule belonging to the immunoglobulin superfamily (IgSF). To take the first step to investigate the role of CD34 in placentation, we examined the expression pattern of CD34 at the maternal–foetal interface in Yorkshire gilts on days 15, 26, 50 or 95 and in Meishan gilts on days 26, 50 or 95 of pregnancy (n = 3 gilts/breed/day of pregnancy) by immunohistochemical technique. The CD34‐positive signals were detected in uterine luminal epithelium and trophectoderm in Yorkshire pigs; the staining for CD34 was located in trophectoderm but barely detectable at the uterine luminal epithelium on day 15 of pregnancy. Then, the expression of CD34 increased dramatically in both the uterine luminal epithelium and trophectoderm by day 26, and weak staining intensity was observed at the maternal–foetal interface on days 50 and 95 of pregnancy. The expression pattern of CD34 in Meishan pigs is similar to that in Yorkshire pigs except that only a few positive signals were observed at the luminal epithelium on day 26 of pregnancy. These results suggest that CD34 may be involved in mediating the cell‐to‐cell adhesion between trophectoderm and the luminal epithelial cells during early pregnancy in pigs.     

Pregnancy‐Associated Glycoprotein (PAG) Profile of Holstein–Friesian Cows as Compared to Dual‐Purpose and Beef Cows

Pregnancy‐associated glycoproteins (PAGs) are produced by mono‐ and binucleate trophoblast cells in the ruminant placenta. PAG appears in maternal blood and, from approximately 4 weeks after fertilization onward, may serve as a reliable means of diagnosing pregnancy. A range of factors are said to affect plasma PAG concentrations, such as number and sex of foetus, mass of calf and placenta, level of milk production and genetic constitution. In this study, PAG pregnancy profiles of a dual‐purpose (Simmental) and two beef breeds (Uckermark and Aubrac) are compared with the profile of the specialized dairy breed Holstein–Friesian. Holstein–Friesian cows were sampled weekly; the levels of the other breeds were presented at 3‐week intervals. The overall significant breed difference (p = 0.013) was founded on deviations during the initial 3 weeks of pregnancy and from 23 weeks onward. During the period critical for the detection of pregnancy, between four and 22 weeks, agreement between PAG levels of various breeds was close (p > 0.05). No significant effect of body mass of cow or calf (relative to mass of dam) was detected. These findings imply that the PAG pregnancy test may be executed uniformly irrespective of breed or type of cow, affirming the suitability of the test as a valuable asset for the cattle industry.     

Pseudo-placentational Endometrial Cysts in a Bitch

Cystic alterations of the canine endometrium compromise reproduction and fertility of the bitch and may lead to life-threatening diseases, such as pyometra. Even without clinical evidence, reduction of the uterine lumen by cysts implicates disturbances during migration, nidation and development of the embryo. Several studies point to the high variability of morphology of uterine endometrial cysts but they lack detailed analyses of alterations. In the present study, immunohistochemistry was used to investigate the expression of steroid hormone receptors (oestrogen, progesterone), proliferation activity, inflammation and infection in the cystic affected tissue regions in contrast to the normal endometrium. Oestrogen receptor expression showed a high density of receptors throughout the surface epithelial cells, crypt epithelial cells, glandular epithelial cells and stromal cells of the normal endometrium as well as the cystic affected regions. Proliferation in the cysts was verified in the middle and basal cells of the crypts. Neither in the endometrium nor in the cysts inflammatory processes or evidence of infection could be detected. Furthermore, lectin histochemistry and electron microscopic methods showed that lectin binding patterns and cell morphology of internal epithelial lining and surface epithelium of the cysts can be used to characterize and distinguish different types of cystic alterations. Analogies between epithelial cells of the glandular chambers of the canine placenta and the cystic cellular morphology, steroid hormone receptor distribution as well as lectin binding patterns of the endometrial cysts, as observed in this study, suggest to introduce the term 'pseudo-placentational endometrial cysts'.     

Use of ultrasonic Doppler waveforms to estimate changes in uterine artery blood flow and vessel compliance

Previous data from our laboratory have demonstrated that uterine blood flow (UBF) and uterine arterial smooth muscle tone vary regularly during the estrous cycle of the cow. Uterine blood flow is highest and uterine arterial tone is lowest at estrus, whereas UBF is lowest and uterine arterial tone is highest during the luteal phase of the cycle. Blood flow through arteries is highly pulsatile; changes in arterial wall properties affect shape of the velocity waveform. This study was conducted to evaluate changes in uterine arterial velocity waveforms throughout the estrous cycle of the cow and to relate these changes to fluctuations in UBF and concentrations of estrogen and(or) progesterone in systemic blood. Pulsatile velocity waveforms were obtained daily from pulsed-wave ultrasonic probes placed surgically on the middle uterine artery of five beef cows exhibiting estrous cycles of normal duration (d 0 = day of estrus). Velocity waveforms varied regularly during the estrous cycle of each cow in association with changes in UBF and steroid concentrations. Further, two distinct velocity waveform shapes were observed during the estrous cycle. The first waveform shape, which was observed during periods of high UBF (d -4 to +4 of the estrous cycle), was characteristic of a highly compliant vessel and was associated with a high estrogen:progesterone ratio. The second waveform shape, which was observed from d 7 to 14 of the estrous cycle, was characteristic of a less compliant vessel and was associated with a depressed estrogen:progesterone ratio. These data suggest that compliance of the uterine artery changes during the estrous cycle in association with the changing estrogen:progesterone ratio in blood.     

Postpartum reproductive function in Finnish Ayrshire and Friesian cows after three subsequent parturitions

The postpartum reproduction functions of 84 Finnish Ayrshire and Friesian cows were studied on an experimental farm. The cows were divided according to type of feed into 2 groups of equal size: a hay-urea group and a silage group. The duration of the study was 3 years. Eighty-four cows were followed during their first postpartum period, 63 during 2 and 57 during 3 consecutive post partum periods.The cows were examined clinically by rectal palpation, and milk samples for progesterone assay were taken 3 times a week between calving and first insemination. External signs of heat were checked 3 times a day by the herdsmen.Intervals from calving to completed uterine and cervical involution, appearance of first palpable follicle, onset of first luteal function and onset of regular ovarian function were recorded. Intervals from calving to the first observed heat and to pregnancy were measured and the rate of successful pregnancies at the first insemination was calculated.Least squares methods were used for evaluation of the effect of individual cow, breed, parity/year and type of feed on postpartum reproductive function.The cows in the herd seemed to have equal reproductive functions which were influenced by the conditions during each year/parity rather than by the breed or type of feed.Individual cow effect was significant for the rapidity of uterine involution. Breed had no significant effect on any of the parameters studied.The effect of type of feed alone was not significant for any parameter, but when studying the intervals from calving to uterine involution and initiation of luteal function differences between the feed groups were noted within parity/year.The percentage of cows showing heat before the first dioestrus (luteal phase) increased with parity/year, and the first dioestrus was shorter than the second. The percentage of cows showing postoestrous bleeding increased with number of oestrous cycles post partum.     

Histological and Histochemical Characterization of the Bovine Rete Ovarii through the Estrous Cycle and Gestation

The rete ovarii of cattle may be morphologically divided into the intraovarian rete, the connecting rete, and the extraovarian rete. The connecting rete may be subdivided into that portion which lies within the ovary proper and that portion that lies in the mesovarium. The portion of the connecting rete which lies in the mesovarium is connected to the infundibulum of the uterine tube by a patent tubo-retial communication.
Mitotic figures were observed in the epithelial cells of the rete ovarii during estrus, during metestrus, following superovulation, and in association with the cystadenomatous change which was observed in one ovary of an estrus cow. The epithelial lining of microcysts appeared to proliferate and produce material which filled the cysts. The occurrence of vesicles in the retial epithelium appeared more commonly during diestrus and gestation. The greatest amount of cells, cellular debris and amorphous luminal material was observed during anestrus, diestrus and gestation.