Post-mortem Examination of Sows Genital Organs Culled for Reproductive Disturbances and Immunohistochemical Studies on ERα and PR A Receptors in the Anoestral Sows Uterus

The aim of present study was post-mortem examination of ovaries, uterus and plasma oestradiol-17beta (E2) and progesterone (P4) concentrations in the blood of sows with reproductive disturbances and the distribution of oestradiol receptor (ERalpha), as well as progesterone (PR-A) in the anoestrous sows uteri. Reproductive organs of 150 crossbred sows (Lithuanian White x Danish Landrace) culled for the reasons of reproductive disturbances, were collected in local abattoir over a period of 3 months (September-November). Organs were assessed to determine the stage of the oestrous cycle or anoestrus and their development. Blood samples were collected for E2 and P4 analysis from the jugular vein 1 h prior to slaughter. For this study uterine samples only from pathological anoestrous sows were subjected to immunohistochemical staining to assess the distribution of ERalpha and PR-A in surface epithelium, subepithelial connective tissue, glandular epithelium and myometrium. Macroscopic examination of the ovaries showed that 68.7% sows had active cycling ovaries, 26.6% sows were anoestrus, ovaries were small without CL, and 4.7% of sows had multiple follicular cysts. In anoestrous sows (n = 27) the number and intensity of the nuclear staining of ERalpha varied between different uterine tissue compartments. The highest number (>80%) and the strongest intensity (+++) of positively stained cells for ERalpha was seen in myometrium and glandular epithelium. In other uterine wall compartments the number and intensity of positively stained for ERalpha nuclei was lower (+/++). The PR-A was absent from all tissue compartments. The intensity of the nuclear staining for ERalpha varied not only between the different uterine compartments but also between the sows. The 11.1% of the sows presented ERalpha in surface epithelium, 74.1% of the sows in glandular epithelium and 63.0% of sows in the myometrium.     

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.     

The Endometrium of the Anoestrous Female Pig: Studies on Infiltration by Cells of the Immune System

The aim of this study was to investigate the distribution of immune cells in the endometrium of anoestrous female pigs, five sows in anoestrus by lactation and five pre-pubertal gilts (Swedish Landrace x Swedish Yorkshire). Uterine samples, taken immediately after slaughter, were fixed, embedded in plastic resin and stained with toluidine blue or cryo fixed and stored in a freezer at -70 degrees C until analysed by immunohistochemistry with an avidin-biotin peroxidase method. Immune cells in the surface (luminal) and the glandular epithelium as well as the subepithelial and the glandular connective tissue layers were counted using light microscopy. In the surface (luminal) and the glandular epithelia of gilts and sows, lymphocytes were the predominant immune cells found. There were no significant differences between gilts and sows. Macrophages were detected in the glandular epithelium of sows but not in gilts. In the subepithelial and the glandular connective tissue layers of both gilts and sows, lymphocytes were also the most common immune cells found. The numbers of lymphocytes and macrophages were significantly higher in the sows than in the gilts (p 相似文献   

Oxytocin receptors in dioestrous and anoestrous canine uteri

The aim of the study was to localize oxytocin receptors (OTR) and measure mRNA expression of OTR in the canine uterus with and without the influence of progesterone. Uterine samples were taken from nine anoestrous and eight dioestrous bitches during ovariohysterectomy. Histological changes were evaluated in haematoxylin and eosin (HE)‐stained samples. Purified polyclonal antibody for OTR was used in immunohistochemistry to localize receptors in uterine layers. Relative mRNA concentration of OTR was evaluated with real‐time PCR from full‐thickness uterine samples taken from the middle horn and the body. Myometrial smooth muscle cells, endometrial luminal epithelium (LE) and deep and superficial glandular epithelium were positively stained for oxytocin receptors in non‐pregnant animals. No significant difference in staining intensity was detected between uterine middle horn and body. However, the staining intensity of LE was significantly higher in dioestrous than in anoestrous uteri (p < .05). Leucocytes and endothelium of blood vessels were also positively stained for OTR. Real‐time PCR showed no significant differences in OTR mRNA expression between the middle horn and the body of the uterus, or between anoestrous and dioestrous uterus. No correlation was noted between OTR mRNA expression and blood progesterone concentration. In conclusion, despite the apparent inactivity, the uterus of the non‐pregnant bitch expresses OTR. The distribution or relative expression of OTR does not differ between uterine horn and body in dioestrus or anoestrus except in LE. LE may have more oxytocin‐dependent activity during dioestrus than anoestrus.     

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.     

Expression of Survivin,Bcl‐2 and Bax Proteins in the Domestic Cat (Felis catus) Endometrium During the Oestrus Cycle

Apoptosis has been shown to be an important regulator of endometrium function. To clarify the regulation of apoptosis in the cat endometrium during the normal oestrus cycle, the expressions of the apoptosis‐related proteins (Bcl‐2 and Bax) and their correlation to the inhibitor of apoptosis protein Survivin were analysed using immunohistochemistry. The TUNEL technique (TdT‐mediated dUTP nick end labelling) was also used to detect DNA fragmentation characteristic of apoptotic cells. The results demonstrated that TUNEL labelling is not effective for the detection of apoptosis in cat endometrium. Survivin was expressed in the luminal and glandular epithelial cells of cat endometrium during all phases of the oestrus cycle. Survivin was localized in both the cytoplasm and nuclei of superficial and deep uterine gland cells during the luteal phase, while only cytoplasmic staining was observed during the follicular and anoestrus phases. Bax immunoreactivity in the cytoplasm of luminal and glandular epithelial cells as well as the smooth muscle cells of blood vessels was weak in the anoestrus phase. Compared with anoestrus, the intensity of Bax immunostaining was moderate in the follicular phase and increased dramatically in the luteal phase. Bcl‐2 immunostaining in the cytoplasm of luminal and glandular epithelial cells was moderate in the anoestrus phase. During the early follicular phase, cytoplasmic Bcl‐2 immunostaining was detected mostly in glandular epithelial cells. In the mid‐follicular phase, in glands, the amount of Bcl‐2 protein increased progressively from the superficial to the deep layer. In contrast, the expression of Bcl‐2 decreased in the secretory phase, being very low or absent in the mid‐ and late luteal phases. The overall results suggest that Survivin, Bax and Bcl‐2 proteins may cooperatively contribute to cell apoptosis and cell proliferation in the cat uterus during the oestrus cycle.     

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.     

Ovarian mucinous cystadenoma with smooth muscle proliferation in a cynomolgus monkey (Macaca fascicularis)

An ovarian mucinous cystadenoma was found in a 5-year-old female cynomolgus monkey (Macaca fascicularis). The tumor was composed of various sizes of multilocular cystic glands lined by a single layer of mucin-filled epithelium. Each of these cystic glands was surrounded by a large amount of solid fibrous stroma resembling smooth muscle. The ovarian surface epithelium showed partial invagination into the ovarian cortex, and a transition was observed between the surface epithelium and the mucinous cyst-forming epithelium. Immunohistochemically, the stromal cells were positive for alpha-smooth muscle actin and proliferating cell nuclear antigen. Ultrastructurally, the glandular epithelium had numerous mucinous secretory granules and microvilli. The stromal cells had numerous parallel microfibrils with focal density. It is rare to encounter evidence of a transition from the surface epithelium to the mucinous tumor epithelium and to show stromal smooth muscle proliferation in a mucinous cystadenoma.     

哺乳动物子宫内膜腺的发育与调控

哺乳动物的子宫内膜腺可合成和分泌或转运一些蛋白及相关物质 ,这些物质在胎儿存活、发育、妊娠识别信号的启动及胚胎着床等方面起重要的作用。子宫内膜腺的发育包括内膜上皮出芽形成内膜腺上皮、内膜腺上皮管穿过固有层及内膜腺上皮卷曲和分支等几个连续的过程。哺乳动物子宫内膜腺的形态发生受许多因素的调控 ,但确切的调控机理目前尚不完全清楚     

Isolation and Culture of Luminal Oviductal and Glandular Uterine Epithelia from the Pig (Sus scrofa)

Contents Pig luminal oviductal and glandular uterine epithelia were isolated and cultured to monolayers. Within 4–5 days after isolation the monolayers from glandular uterine epithelium reached confluency, while cultures of luminal oviductal epithelium needed 6–7 days to reach this stage. The epithelial character of the cultured cells was confirmed by scanning electronmicroscopy (SEM) and by demonstration of intermediate filaments using immunohitochemistry. SEM analysis revealed that the monolayers from luminal oviductal origin consisted of microvilli rich epithelial cells only. In SEM preparations from glandular luterine epithelium we observed microvilli bearing as well as ciliated epithelial cells. All isolates from glandular uterine epithelium contained connective tissue derived fibroblasts. In cryosections of fresh oviductal and endometrial tissues, epithelial cells did react specifically with anti-keratin while anti-vimentin was not bound. Cultured epithelial cells, however, reacted with both anti-vimentin and anti-keratin indicating that cells from oviductal or urterine origin might dedifferentiate with respect to intermediate filament expression when brought into culture. Inhalt: Isolierung und Kultur von Epithelzellen aus dem Eileiterlumen und von glandulären uterinen Epithelzellen beim Schwein (Sus scrofa) Porcine Epithelzellen aus dem Eileiterlumen und glanduläre uterine Epithelzellen wurden isoliert und als Monolayer kultiviert. Innerhalb von 4–5 Tagen nach der Isolierung waren die uterinen Epithelzellen zusammengewachsen. Epithelzellen des Eileiters benötigten 6–7 Tage um dieses Stadium zu erreichen. Der epitheliale Charakter der Zellen wurde durch elektronenmikroskopische Untersuchungen (SEM) und durch den immunhistologischen Nachweis intermediärer Filamente bestätigt. DieelektronenmikroskopischeUntersuchung zeigte, daβdie Monolayer aus den Eileiterzellen nur aus Mikrovilli-reichen Epithelzellen bestanden. Dagegen wiesen die glandulären Uterinzellen sowohl Zellen mit Mikrovilli als auch mit Zilien besetzte Epithelzellen bei der elektronenmikroskopichen Untersuchung auf. Alle isolierten Epithelzellen uterinen Ursprungs enthielten Fibroblasten aus dem verbindenden Gewebe. In Cryoschnitten frischer Eileiter- und Endometriumsgewebe reagierten die Epithelzellen spezifisch mit Anti-Keratin, wohingegen Anti-Vimentin nicht gebunden wurde. Epithelzellen aus der Kultur reagierten jedoch mit beiden Substanzen. Dies kann als Hinweis gewertet werden, daβ die Zellen in der Kulturphase in Abhängigkeit der Intermediärfilamente entdifferenziert werden.     

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.     

Expression and Hormonal Regulation of E‐Cadherin in Canine Uterus During Early Pregnancy

E‐cadherin, a Ca^{2 +} ‐dependent cell adhesion molecule, is necessary for endometrial receptivity to blastocyst implantation. The aim of this study was to investigate the differential expression of E‐cadherin in canine uterus during early pregnancy and its regulation under different conditions by in situ hybridization. E‐cadherin mRNA expression was at a low level in the glandular epithelium on days 6, 12 and 17 of pregnancy. On days 20 and 23 of pregnancy, E‐cadherin mRNA was highly expressed in the glandular epithelium surrounding the embryo, but not in the luminal epithelium and declined in villi and placenta on day 28 of pregnancy. During oestrous cycle, a moderate level of E‐cadherin mRNA expression was found in the luminal and glandular epithelium of canine uteri at oestrus stage. The same expression was also found at anoestrus stage. Progesterone slightly induced the expression of E‐cadherin mRNA in the luminal and glandular epithelium of ovariectomized canine uterus. These results suggest that E‐cadherin expression is closely related to canine implantation and can be up‐regulated by progesterone.     

Immunohistochemical Studies on Oestrogen Receptor Alpha (ERα) and the Proliferative Marker Ki-67 in the Sow Uterus at Oestrus and Early Pregnancy

Oestrogen receptor alpha (ERalpha), the main subtype in the uterus, is involved in the regulation of uterine growth/proliferation. A relationship between ERalpha and proliferative activity has been shown in the cyclic sow uterus, but to our knowledge, no study has been carried out on early pregnant sows. Therefore, by means of immunohistochemistry and use of mouse monoclonal antibodies to ERalpha and a proliferative marker, Ki-67, the localization of these proteins was investigated in the sow uterus during early pregnancy. Eighteen crossbred multiparous sows were artificially inseminated once at 20-15 h before expected ovulation. After artificial insemination (AI), they were slaughtered at five different times: at oestrus, 5-6 h after AI (n = 4), 20-25 h after ovulation (n =4), 70 h after ovulation (n = 4), on day 11 (the first day of standing oestrus = day 1, n = 3) and on day 19 (n = 3). Immediately after slaughter, uterine samples were collected at the mesometrial side of the uteri, fixed in 10% formaldehyde and embedded in paraffin. Immunohistochemistry was performed by using mouse monoclonal antibodies to ERalpha (C-311) and Ki-67 (MM1). All sows slaughtered after ovulation were pregnant. In general, positive immunostaining for ERalpha and Ki-67 was found in the nuclei. Variations in staining intensity and proportion of positive nuclei were observed in different uterine compartments and stages of early pregnancy. The highest level of ERalpha presence in the surface epithelium and myometrium was found at oestrus (5-6 h after AI), and low levels of ERalpha in these compartments were observed as early as 20-25 h after ovulation. In the glandular epithelia, presence of ERalpha was highest at 70 h after ovulation. The largest number of ERalpha-positive cells in the stroma was observed at oestrus and early after ovulation. Low proliferation was observed, and with no significant difference in tissue compartments except in the glandular epithelium. High proliferative activity in the glandular epithelium at 70 h after ovulation indicated involvement in preparation for secretory activity and growth during pregnancy establishment. Significant positive correlation was found between the number of ERalpha-positive cells in the stroma and Ki-67-positive cells in the surface epithelium. In conclusion, the present study showed differences in immunolocalization of ERalpha and the proliferative marker Ki-67 in different tissue compartments of the sow uterus at oestrus and early pregnancy. In some uterine compartments, the patterns of ERalpha and Ki-67 immunostaining seemed to be influenced by insemination and the presence of embryos, in addition to the effects of steroid hormones.     

山羊子宫内胆碱能神经分布及妊娠时的变化

采用乙酰胆碱酯酶组化法，研究了山羊子宫内胆碱能神经的分布，结果，山羊子宫颈是经较丰富，在浆膜和肌层内有神经束伴血管而行并分支分布于血管壁，在粘膜及其皱褶上皮下，粘液腺上皮有神经丛分布，妊娠时子宫颈部的神经分布与未妊娠时相比无明显变化，子宫角部神经密度均低于子宫颈部，其内环行肌层中及其与内膜交界处神经密度略高，神经束伴血管而行并分支分布于血管壁，在子宫腺上皮下及内膜上皮下无神经分布，妊夺时作胎盘内无神经分布外，仍有神经束伴血管而行交分支分布于血管壁，在分布于血管壁的神经纤维减少，结果提示，胆碱能神经主要支配山羊子宫内血管壁及颈部粘液腺上皮和粘膜上皮，妊 时子宫颈部胆碱能神经无明显变化，而子宫角内支配血管壁的胆碱能神经纤维减少。     

Sialomucin Complex (Muc4) Expression in Porcine Endometrium During the Oestrous Cycle and Early Pregnancy

The non‐invasive type of implantation in the pig is characterized by the maintenance of a thick glycocalyx coating on the uterine epithelial surface microvilli. Present study investigated the alteration in the sialomucin complex (Muc4) expression during the oestrous cycle and early pregnancy in the pig. Endometrial tissue samples were immunostained with the primary antibody to the Muc4 transmembrane subunit ASGP‐2. Muc4 immunostaining increased in the surface and glandular epithelia between days 5 and 10 of oestrous cycle. Immunostaining continued to increase on day 12 with the greatest intensity of uterine Muc4 immunostaining detected on day 15 of the oestrous cycle and early pregnancy. Endometrial Muc4 expression in cyclic gilts decreased dramatically during early proestrous but continued to remain abundant in the surface and glandular epithelium of pregnant gilts during the period of conceptus attachment to the uterine surface.     

X-ray Microanalysis of the Secretory Epithelium of the Endometrial Glands and Intraluminal Uterine Fluid in Oestrus Mares

X‐ray microanalysis was used to determine the occurrence of transudation as a source of intrauterine fluid in mares during oestrus. The content of some selected elements was studied in secretory vesicles of the epithelium of endometrial glands and in intraluminal fluid of healthy mares of varying age and parity. An in situ cotton tampon was used to absorb intrauterine fluid from 23 gynaecologically healthy mares during oestrus. Immediately after tampon withdrawal, endometrial tissue for biopsy was removed from the uterine body/horn junction while videoendoscopy was performed. The tissue specimens were frozen ultra‐rapidly against a copper surface chilled with liquid N₂. X‐ray microanalysis of frozen fully hydrated endometrial samples and of microdroplets of intraluminal fluid (n = 20) was performed, using a SEM microscope equipped with a cryo‐stage and an X‐ray microanalysis system. The concentrations of sodium (Na), potassium (K), chlorine (Cl), sulphur (S), phosphorus (P) and calcium (Ca) in the epithelium of endometrial glands and in intraluminal fluid were compared with those in serum. There was a significant difference in the concentrations of all analysed elements between the secretory gland epithelium and the intraluminal fluid apart from S. The major elements in glandular epithelium were K and P. Although there was a significant difference in the concentrations of Na, Cl, S and Ca between the uterine fluid and serum, there was still a conspicuous resemblance in the relationship between the different elements in uterine fluid and serum, with Na and Cl being the dominant elements. Sulphur and especially Ca were present only in small amounts throughout. Neither age nor parity affected the elemental concentrations of Na, K, Cl, S, P and Ca in gland epithelium, uterine fluid or serum. It was concluded that serum transudation contributes to a great extent to the composition of uterine fluid, having a diluting effect upon the specific secretion of the uterine glands.     

犬正常发情期与患子宫蓄脓的子宫和卵巢组织结构及乳铁蛋白表达的对比

本研究旨在探明犬子宫和卵巢在正常发情期与患子宫蓄脓时，其组织结构及乳铁蛋白（lactoferrin，LF）表达的变化特征。应用Masson’s、VVG、PAS组织化学染色方法观察乏情期、发情期、患子宫蓄脓时犬子宫及卵巢的组织结构特点，用免疫组织化学SP法观察LF的分布特征。结果显示，正常发情期犬：乏情期、发情期子宫内膜肌层厚度比分别为0.762 0、0.924 3；乏情期子宫固有层中胶原纤维含量大于发情期，发情期卵巢中胶原纤维含量大于乏情期；子宫血管层及卵巢血管内弹性膜清晰完整；子宫腺在乏情期时浅层管腔小，深层较大、腺管上皮为单层柱状上皮、上皮细胞及管腔内PAS阳性反应较强，发情期子宫腺管腔变大、腺管上皮为单层立方上皮、上皮细胞及管腔内有PAS阳性反应；子宫黏膜上皮在乏情期和发情期均为单层柱状上皮，但胞核位置不同，乏情期胞核位于中央，发情期胞核位于顶部。患子宫蓄脓犬：子宫内膜肌层厚度比为1.615 0；子宫固有层和卵巢中胶原纤维含量少于正常发情期；子宫腺管腔大，且形状不规则，管腔内有炎性细胞浸润，腺管上皮为单层立方上皮，有淋巴细胞位于基膜，上皮细胞及管腔内PAS阳性反应较弱；子宫血管层及卵巢血管内弹性膜较正常发情期变薄，且有断裂现象；子宫黏膜上皮为单层柱状上皮，胞核位于基底。LF在乏情期子宫腺上皮和卵巢中的表达水平高于发情期，而在子宫黏膜上皮中发情期的表达水平高于乏情期。患子宫蓄脓时犬子宫和卵巢中LF的表达水平均较低。综上表明，犬正常发情期与患子宫蓄脓时的子宫和卵巢组织结构特点显著不同，LF的表达水平也存在差异。     

Immunohistochemical Detection of Receptors for Oestrogen and Progesterone in Endometrial Glands and Stroma during the Oestrous Cycle in Nelore (Bos taurus indicus) Cows

The aim of the present study was to monitor endometrial distribution and concentrations of oestrogen receptors α (ERα) and progesterone receptors (PR) by immunohistochemistry in Nelore cows (Bos taurus indicus) during the oestrous cycle. Blood samples were collected for progesterone measurement and endometrial samples were taken from the uterine horn contra lateral to the corpus luteum in 16 cows at days 0 (ovulation), 5, 9, 13 and 19 of the oestrous cycle. Immunostaining evaluation for ERα and PR in the glandular epithelium and uterine stroma was performed by two methods: positive nuclei counting and staining intensity of the nuclei. Specific positive staining reactions for both receptors were limited to cell nuclei and they were not identified in the cytoplasm. The proportion of ERα positive nuclei had a temporal variation throughout the oestrous cycle in both cell types evaluated and was higher in uterine stroma than the glandular epithelium (p < 0.05). The greatest proportion of ERα stained nuclei was observed at oestrus and during the initial and mid luteal phase (days 5, 9 and 13) (p < 0.05) in the glandular epithelium and at days 0, 5 and 9 in the uterine stroma (p < 0.01). The proportion of PR positive nuclei remained constant throughout the entire oestrous cycle for both cell types evaluated (p > 0.05). A higher proportion of PR positive nuclei was measured in the uterine stroma compared with the glandular epithelium (p < 0.05). Intensity of staining for ERα and PR varied throughout the oestrous cycle (p < 0.01). There was a higher staining intensity at days 0 and 5 in the stroma for ERα (p < 0.01) and PR (p < 0.01) and in the glandular epithelium at days 0, 5, 9 and 13 for ERα (p < 0.01) and at days 0, 5 and 9 for PR (p < 0.01) when compared with the other evaluated days. These data demonstrate that ERα and PR expression varied throughout the oestrous cycle in Nelore cows, in general with highest concentrations at oestrus and the lowest during the luteal phase. This is similar to patterns observed in Bos taurus taurus.     

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'.