ABSTRACT
In porcine placenta, abnormal development of the placental vasculature leads to placental insufficiency. The aim of this study was to determine the mRNA expression of angiogenic growth factors and to determine the vascular characteristics in placenta at day 40 of pig gestation. Samples were collected from maternal-chorioallantoic interface (n = 21) for the measurement of mRNA expression of VEGFA, ANGPT1, ANGPT2, FGF2 and its receptors KDR, TEK, FGFR1IIIc, FGFR2IIIb respectively, and for immunohistochemistry analysis of CD31 and VEGFA. Immunohistochemical analysis of CD31 and VEGFA, morphometric measurement of blood vessels, high-resolution light microscopy and transmission electron microscopy were performed. Capillary area density, number of blood vessels and capillary area were significantly higher on the maternal side than on the fetal side (p < .05). The ultrastructural finding of blood vessels demonstrates close contact with the trophoblastic epithelium. The relative mRNA expression of VEGFA and its receptor KDR was higher compared with the other angiogenic genes. In conclusion, a high mRNA expression of VEGFA and its receptor KDR added to the immunohistochemical results suggest a potential role of these genes in this pathway associated with an increase in the density of the capillary area on the maternal side and a reduction in the hemotrophic diffusion distance at the interface for nutrient exchange.
Subject(s)
Placenta , Trophoblasts , Pregnancy , Female , Animals , Swine , Placenta/metabolism , Fetus/blood supply , Morphogenesis , RNA, Messenger/metabolismABSTRACT
Bovine tritrichomonosis, a sexually transmitted disease caused by the protozoan Tritrichomonas foetus, is characterized by producing reproductive alterations in cattle. Carbohydrates on the surface of the uterine epithelium are involved in the process of adhesion and colonization of the protozoan. The murine model has proved to be an inexpensive, practical and representative alternative to study the lesions produced in the natural host. For this work, during the first stage, 6-8 week old female BALB/c mice were inoculated with 24 different T. foetus isolates in order to classify them according to their pathogenicity. Then, seven isolates were selected and processed with lectin histochemistry to determine if the differences in pathogenicity corresponded to the changes found in the uterine carbohydrate expression pattern. In this work, we demonstrate the differences in the expression of the carbohydrate pattern between infected and uninfected mice. In addition, within the group of infected mice, differences were found in the degree of pathogenicity of the isolates, thus evidencing their biological variability.
Subject(s)
Cattle Diseases , Protozoan Infections, Animal , Rodent Diseases , Tritrichomonas foetus , Cattle , Animals , Female , Mice , Mice, Inbred BALB C , Uterus/pathology , Cattle Diseases/metabolism , Carbohydrates , Protozoan Infections, Animal/metabolism , Protozoan Infections, Animal/pathologyABSTRACT
The liver has multiple functions that change throughout ontogeny. South American camelids (SAC) have unique characteristics related to adaptation to extreme environments and metabolism. However, the process of hepatic cell differentiation has not been studied in any SAC. We study the patterns of cell differentiation and proliferation in the liver of the alpaca at different times of the ontogeny, excluding the hematopoietic components. Immunohistochemical techniques were performed in 66 specimens, including embryos, fetuses, neonates and adults. Supplementary analyses were performed by lectinhistochemistry. The hepatocytic differentiation was performed by the identification of Hepatocyte (Clone: ââOCH1ES Dako®). It began in the specimens of 1.8-2.5 cm of crown to rump length (CRL), from Days 25-29 (ovulation = Day 0), continued during gestation and intensified towards its end. The cholangiocytic differentiation was performed by the identification of cytokeratin 7 (CK7, Dako®). It was manifested at the final of gestation (specimens of 28.4 cm CRL, from Day 223 onwards). Parenchymal cells underwent a process of gradual differentiation (differentiation of hepatocytes preceded that of cholangiocytes). Cell proliferation was observed along gestation using the nuclear proliferation antigen (PCNA) and Ki-67. Hepatic organogenesis in the alpacas shares similar differentiation and proliferation mechanisms with other altricial, but phylogenetically distant, species.
Subject(s)
Antigens, Differentiation/metabolism , Camelids, New World/embryology , Cell Differentiation , Cell Proliferation , Hepatocytes/metabolism , Liver/embryology , Animals , Female , MaleABSTRACT
Seals (Phocidae) undergo an annual cycle of moulting that implies hair regeneration, and in the case of southern elephant seals, it also involves the superficial strata of the epidermis. Therefore, surviving the moulting period is crucial for their obligate and permanent ectoparasites. Throughout evolutionary time, sucking lice (Echinophtiriidae) have developed morphological, behavioural and ecological adaptations to cope with the amphibious lifestyle of their hosts. Lepidophthirus macrorhini, the Southern elephant seal louse species, faces the additional challenge of surviving attached to the host during the moulting period. Since lice live on the skin, L. macrorhini has developed a unique survival strategy by piercing the skin of their host, thus keeping them protected from moulting. During fieldwork in Patagonia and Antarctica, skin samples with lice within were collected for histological analysis to assess whether these parasites caused damage to the host. Lice generate an inflammatory process in the host's dermis, and these lesions could alter the normal chemical and mechanical protective properties of the skin facilitating secondary infections. Further studies that analyse the potential pathogens in those skin lesions are necessary to properly assess the real impact of ectoparasites on their host health.
Subject(s)
Anoplura , Seals, Earless , Animals , Molting , Seals, Earless/parasitology , Seals, Earless/physiology , SkinABSTRACT
Cutaneous papillomas (CPs) and cutaneous squamous cell carcinomas (CSCCs) are usual epidermal tumours in dogs. CPs and CSCCs probably arise from the neoplastic transformation of the keratinocytes within the stem cell compartment, since these cells are the only keratinocytes that would reside long enough to accumulate the number of molecular alterations to drive the progression towards a tumour cell phenotype. However, the role of these cells in common epidermal tumours in dogs is still unknown. Thus, the purpose of this study was to evaluate the immunohistochemical expression pattern of p63 together with CK5, molecular markers of epidermal stem cells, on sections of tissue microarrays constructed from canine samples of CP and CSCC to investigate the contribution of stem cells in those canine tumours. p63/CK5 coexpression was retained in most basal and some suprabasal cells in CPs and CSCCs. In addition, increased coexpression of these molecules was observed in a group of CPs and CSCCs, as a result of a higher p63 expression. These results suggest that the coexpression of p63/CK5 may mark epidermal keratinocytes that possess self-renewal capacity rather than only stem cells, and suggest that transit amplifying cells, and even differentiated keratinocytes, may also contribute to the pathogenesis of epidermal tumours in dogs.
Subject(s)
Carcinoma, Squamous Cell/veterinary , Dog Diseases/metabolism , Papilloma/veterinary , Skin Neoplasms/veterinary , Animals , Biomarkers, Tumor/metabolism , Carcinoma, Squamous Cell/genetics , Dogs , Papilloma/genetics , Skin Neoplasms/pathologyABSTRACT
The aims of this study were: to ultrasonograhically describe and compare testicular parenchyma echogenicity and heterogeneity using digital image analysis in: I) prepubertal (PREP), peripubertal (PERI) and mature (MAT) cats; II) Normal and abnormal mature felids. Secondary, the relationships between histomorphological and ultrasonographic attributes of the testes were also determined. I) Fourteen, PREP, PERI and MAT male cats were ultrasonographically examined and then castrated. II) Seven adult cats were ultrasonographically examined before and after a GnRH antagonist administration and then castrated. All the testes were grossly and histomorphometrically assessed. In the frozen digital images of the longitudinal ultrasound sections, 3 regions of interest (ROI, 1 mm2) were selected. Within each ROI the echogenicity and the heterogeneity of the testicular parenchyma were digitally analyzed. In experiment I, testicular volume (0.15 ± 0.0 vs. 0.49 ± 0.1 vs. 1.65 ± 0.1; P < 0.01) and gonadosomatic index (0.04 ± 0.0 vs. 0.05 ± 0.0 vs. 0.08 ± 0.0; P < 0.01), echogenicity (56.54 ± 0.75 vs. 81.87 ± 5.88 vs.94.67 ± 3.62; P < 0.01) and heterogeneity (10.2420 ± 1.3740 vs.13.65 ± 0.65 vs. 14.67 ± 1.49; P < 0.01) augmented throughout PRE, PERI, and MAT. In experiment II, testicular volume (1.00 ± 0.09 vs. 0.85 ± 0.09; P < 0.05), echogenicity (87.74 ± 1.53 vs. 83.32 ± 1.54; P 0.01) but not heterogeneity (14.09 ± 0.26 vs. 14.19 ± 0.29; P > 0.05) decreased in the post GnRH antagonist abnormal testes. For both experiments, testicular volume, seminiferous tubular diameter, percentage of spermatids as the most mature cell type, and luminal/intertubular ratio were highly correlated (P < 0.01) with their echotextural attributes. Computer-assisted image analysis of B mode ultrasonogram appears as a good indicator of pubertal development and mild alterations of spermatogenesis in felids.
Subject(s)
Spermatogenesis , Testis , Animals , Cats , Image Processing, Computer-Assisted , Male , Spermatids , Testis/diagnostic imaging , Ultrasonography/veterinaryABSTRACT
Clinically relevant epidermal tumours in dogs include cutaneous papillomas (CPs) and cutaneous squamous cell carcinomas (CSCCs). The development of CPs and CSCCs involves dysregulation in expression of E-cadherin/ß-catenin; however, knowledge about the contribution of these molecules to epidermal tumourigenesis in dogs is limited. This study examined the immunohistochemical expression pattern of E-cadherin/ß-catenin in samples of normal canine epidermis, CPs, preneoplastic epidermis and CSCCs, using tissue microarrays, in order to elucidate whether the dysregulated expression of these molecules may contribute to the pathogenesis of clinically relevant epidermal tumours in dogs. We also investigated the correlation between the immunohistochemical expression pattern of E-cadherin/ß-catenin in these tissue microarrays to further evaluate whether the disruption of the adherens junction interactions plays a relevant role in canine epidermal tumourigenesis. In samples of CP and preneoplastic epidermis, the membrane immunoreactivity of E-cadherin/ß-catenin was conserved, while in CSCC, the immunoreactivity of these molecules was significantly reduced, independently of the tumour location. There was significant correlation between the membrane expression of E-cadherin/ß-catenin in CSCC. ß-catenin also showed cytoplasmic and nuclear expression in samples of CP, preneoplastic epidermis and CSCC. These results support the hypothesis that dysregulated expression of E-cadherin/ß-catenin may play a critical role in the pathogenesis of relevant canine epidermal tumours, not only due to the disruption of the intercellular adherens junctions, but also due to the dysregulated activity of the signalling pathways in which these molecules are involved.
Subject(s)
Cadherins/metabolism , Dog Diseases/metabolism , Dog Diseases/pathology , Skin Neoplasms/veterinary , beta Catenin/metabolism , Animals , Carcinogenesis/metabolism , Carcinogenesis/pathology , DogsABSTRACT
Zika virus (ZIKV) infection during pregnancy is associated with a spectrum of developmental impairments known as congenital Zika syndrome (CZS). The prevalence of this syndrome varies across ZIKV endemic regions, suggesting that its occurrence could depend on cofactors. Here, we evaluate the relevance of protein malnutrition for the emergence of CZS. Epidemiological data from the ZIKV outbreak in the Americas suggest a relationship between undernutrition and cases of microcephaly. To experimentally examine this relationship, we use immunocompetent pregnant mice, which were subjected to protein malnutrition and infected with a Brazilian ZIKV strain. We found that the combination of protein restriction and ZIKV infection leads to severe alterations of placental structure and embryonic body growth, with offspring displaying a reduction in neurogenesis and postnatal brain size. RNA-seq analysis reveals gene expression deregulation required for brain development in infected low-protein progeny. These results suggest that maternal protein malnutrition increases susceptibility to CZS.
Subject(s)
Malnutrition/complications , Zika Virus Infection/congenital , Zika Virus Infection/complications , Animals , Animals, Newborn , Body Weight , Brain/enzymology , Brain/pathology , Brazil/epidemiology , Diet, Protein-Restricted , Disease Outbreaks , Embryo, Mammalian/pathology , Female , Gene Expression Regulation, Developmental , Malnutrition/virology , Mice, Inbred C57BL , Microcephaly/complications , Microcephaly/virology , Neurogenesis , Organ Size , Pregnancy , Syndrome , Viral Load , Zika Virus Infection/virologyABSTRACT
The molecular mechanisms contributing to the development of cutaneous papillomas (CPs) and cutaneous squamous cell carcinomas (CSCCs) are still poorly understood, limiting the ability to identify molecular suitable targets for the development of novel therapies. Persistent activation of the phosphatidylinositol 3-kinase/Akt/mammalian target of rapamycin (PI3K/Akt/mTOR) signalling pathway is a component of epidermal carcinogenesis in dogs. The present study describes the immunohistochemical expression pattern of two key regulatory molecules involved in the PI3K/Akt/mTOR signalling pathway, phosphorylated epidermal growth factor receptor (pEGFR)Tyr1068 and phosphatase and tensin homologue (PTEN), in samples of normal canine epidermis, CP, preneoplastic epidermis and CSCC using tissue microarrays to determine whether the deregulated activity of these molecules is involved in the pathogenesis of these relevant epidermal tumours of dogs. Expression of pEGFR and PTEN was dysregulated in most samples of CP, preneoplastic epidermis and CSCC. Overexpression of pEGFR, together with decreased expression of PTEN, may facilitate the progression of some canine CPs and CSCCs by deregulation of the key cellular functions in which the PI3K/Akt/mTOR signalling pathway is involved. These findings suggest that the PI3K/Akt/mTOR signalling molecules may be potential therapeutic targets for canine patients with CP and CSCC.
Subject(s)
Carcinoma, Squamous Cell/veterinary , Dog Diseases/metabolism , ErbB Receptors/metabolism , PTEN Phosphohydrolase/metabolism , Papilloma/veterinary , Skin Neoplasms/veterinary , Animals , Carcinogenesis/metabolism , Carcinoma, Squamous Cell/metabolism , Dogs , Papilloma/metabolism , Phosphorylation , Skin Neoplasms/metabolismABSTRACT
The aim of this study was to describe the seminal, histomorphological and hormonal effects of the oral indenopyridine RTI-4587-073(l) on feline testicle. Clinical side effects were also recorded. Sixty testicles of 30 adult cats that had been treated (d 0) with RTI-4587-073(l) 12.5 mg/kg PO and randomly hemiorchiectomized twice on: day -14 (n = 8), 6 h (n = 6), 12 h (n = 8), 24 h (n = 6), day 7 (n = 8), day 14 (n = 6), day 21 (n = 6), day 35 (n = 6) or day 42 (n = 6) were studied. Before each hemiorchiectomy, fecal samples for testosterone (T) measurement were collected and the testes were grossly and ultrasound examined. This indenopyridine did not cause changes in testicular weight (P > 0.1), volume (P > 0.1), echostructure, gonadosomatic index (P > 0.1), fecal T concentrations (P > 0.1), nor clinical side effects. A severe disorganization of the cytoarchitecture of the seminiferous epithelium, sloughed cells and fluid, were observed in the 6 h samples up to a maximum at 24 h. Tubular diameter (P < 0.01) increased twice, during the first 24 h and on d 35. Germinal epithelium achieved its minimum height on d 14 to rapidly recover thereafter. This treatment caused a significant decrease in the volume of all the seminiferous cell components, except spermatogonias. All histotological parameters normalized by the end of the study. It was concluded that RTI-4587-073(l) severely disrupted spermatogenesis during the first 24 h after treatment returning to normality in approximately one spermatic cycle without clinical side effects.
Subject(s)
Cats , Contraceptive Agents, Male/pharmacology , Indenes/pharmacology , Orchiectomy/veterinary , Piperidines/pharmacology , Testis/drug effects , Animals , Male , Orchiectomy/methods , Random Allocation , Seminiferous Epithelium/drug effects , Sperm CountABSTRACT
Cutaneous squamous cell carcinoma (cSCC) represents one of the most common malignant skin tumors in dogs. Research aimed at clarifying how the deregulated activity of signalling pathways contributes to cSCC progression can help to identify molecular suitable targets for the development of novel therapies. The present study describes the immunohistochemical expression pattern of two proteins (pAktSer473 and pS6Ser235/236, the latter combined with Ki-67) involved in the phosphatidylinositol 3-kinase/Akt/mammalian target of rapamycin (PI3K/Akt/mTOR) signalling pathway in canine specimens of normal epidermis, pre-neoplastic epidermis, and cSCC using tissue microarrays. The results suggest that the PI3K/Akt/mTOR signalling pathway has a low expression in the normal canine epidermis, and that selected molecules involved in this signalling pathway are dysregulated during the canine epidermal carcinogenesis process. These findings provide important evidence that the persistent activation of the PI3K/Akt/mTOR signalling pathway represents one of the key events during cSCC progression in canine patients, pointing to the PI3K/Akt/mTOR pathway as a potential therapeutic target.
Subject(s)
Carcinoma, Squamous Cell/veterinary , Dog Diseases/metabolism , Immunohistochemistry/veterinary , Phosphoproteins/analysis , Skin Neoplasms/veterinary , TOR Serine-Threonine Kinases/metabolism , Animals , Carcinoma, Squamous Cell/chemistry , Dogs , Phosphatidylinositol 3-Kinase/metabolism , Proto-Oncogene Proteins c-akt/metabolism , Signal Transduction/physiology , Skin Neoplasms/chemistryABSTRACT
The aim of this study was to describe the lectin-binding pattern in the placentas of cows infected experimentally with Neospora caninum. Four cows were inoculated intravenously with 1 × 108 tachyzoites of the NC-1 strain of N. caninum at 150 ± 7 days of pregnancy. Two control cows were administered a placebo. An indirect fluorescence antibody test (IFAT) was performed on serum samples obtained before and after the inoculation. The cows were killed at 30 and 37 days post inoculation. Samples of placenta were taken for histopathology and lectin histochemistry. Fetal tissues and fluids were collected for histopathology and IFAT, respectively. All infected cows had high antibody titres. All fetuses had characteristic histopathological lesions, including non-suppurative meningoencephalitis, myocarditis, hepatitis and myositis, suggesting N. caninum infection. Only two infected fetuses developed specific antibodies. Mild non-suppurative inflammatory infiltrates were recorded in the placentae. Differences in the lectin-binding pattern were observed between infected animals and controls in the glycocalyx (CON-A and WGA) and apical cytoplasm (RCA-I and CON-A) of the trophoblastic cells; giant trophoblastic cells (CON-A and DBA); glycocalyx (PNA, WGA) and apical cytoplasm (CON-A, WGA, PNA, DBA and RCA-I) of endometrial cells; trophoblast of the interplacentomal region (WGA); endothelium (CON-A, SBA, RCA-1 and WGA); and finally, mesenchyme (CON-A, RCA-1, SBA, PNA and DBA). These findings indicate that there is a distinctive pattern of lectin binding in the placenta of cattle infected with N. caninum. The direct effect of the presence of the protozoa as well as the altered expression of cytokines could explain these changes in the maternofetal interface.
Subject(s)
Cattle Diseases/metabolism , Coccidiosis/veterinary , Lectins/analysis , Placenta/metabolism , Animals , Cattle , Female , Fluorescent Antibody Technique, Indirect , Neospora , Placenta/microbiology , PregnancyABSTRACT
Feline injection site sarcomas (FISSs) are mesenchymal neoplasms that develop at the sites of delivery of vaccines or other injectable products. Vaccine adjuvants can trigger an intense and persistent inflammatory response that may lead to neoplastic transformation. The proinflammatory role of cyclo-oxygenase (COX)-2 is well known and its overexpression has prognostic value in multiple neoplastic processes. One hundred and seventeen FISSs were evaluated for the degree of inflammation and anaplasia. Immunohistochemistry was used to determine the expression of COX-2 in these sarcomas. There was a significant association between the degree of inflammation and the expression of COX-2 by neoplastic cells. COX-2 expression was lower in tumours with higher degrees of anaplasia. These findings may be useful in predicting the sensitivity of FISSs to treatment with COX-2 inhibitors. The potential therapeutic use of such agents could then be restricted to tumours with lower degrees of anaplasia.
Subject(s)
Cat Diseases/etiology , Cat Diseases/pathology , Injection Site Reaction/veterinary , Sarcoma/veterinary , Soft Tissue Neoplasms/veterinary , Anaplasia/veterinary , Animals , Cat Diseases/metabolism , Cats , Cyclooxygenase 2/metabolism , Inflammation/veterinaryABSTRACT
The breeding of South American camelids is the main economic activity of the high Andean region of South America and it, is potentially, the most profitable resource in of the Puna environmental conditions of the Puna. The duration of the gestation in alpaca is 339.7 ± 12 days. The objective of the present work was to macroscopically and microscopically describe the ontogenic development of the splanchnic cavities of the alpaca and to determine the gestational time in which the post-cranial ossification centers are observed in the embryos/fetuses of this species, from day 21 to 107 of gestation. The documentation of normal ontogenic development, which is vacant for this period, is of the utmost importance to understand the consequences of the alterations at the different gestational times, as well as for the estimation of the gestational age in the case of abortions. Forty-seven alpaca specimens of both sexes, at different times of their gestational development, collected during slaughter at local slaughterhouses of the Department of Huancavelica, Peru, were evaluated. Specimens were assigned to seven groups according to their morphological characteristics. The embryogenesis in the alpaca was characterized by a series of changes comparable to those occurring in other mammals with similar gestational periods. Despite these similarities, species differences were found in some organs as stomach, which are observed too in adult individuals.
Subject(s)
Camelids, New World/embryology , Embryo, Mammalian/anatomy & histology , Animals , Embryonic Development , Female , Gestational Age , Male , Osteogenesis , Pregnancy , Stomach/embryologyABSTRACT
Mammalian pregnancy requires specific interactions between the conceptus and its mother that involve the endocrine system and adhesion molecules. The relation between adhesion molecules and their ligands at the fetal-maternal interface is crucial for developing a successful implantation. Progesterone (P4) and estrogen (E2) secreted by the porcine conceptus are required for the relation to be established. We investigated the expression of αvß3 integrin and its ligand, fibronectin (FN), at the placental interface, and E2 and P4 concentrations in both serum and maternal and fetal placental extracts during placentation in swine. Placental and serum samples of crossbred sows at 17, 30, 60, 70, and 114 days gestation and no pregnant uteri were used. The presence of αvß3 and FN were determined by immunohistochemistry, and E2 and P4 by chemiluminescence in homogenates of nonpregnant uterus (HoU), swine maternal placenta (HoPM), swine fetal placenta (HoPF) and serum. The expression of αvß3 and FN increased at the interface at 17, 30 and 60 days gestation. Immunostaining decreased by 70 days. Serum E2 levels peaked at 17 days, then decreased, then increased again near term. The highest concentration of P4 occurred in HoPF at 70 days gestation, then decreased coincident with a decline in integrin and FN expression at the placental interface. High P4 levels during swine gestation may regulate the expression of αvß3 integrin and FN at the placental interface for up to 70 days gestation. Other adhesion molecules and their ligands likely maintain the fetal-placental interface after 70 days.
Subject(s)
Estrogens/pharmacology , Fibronectins/metabolism , Integrin alphaVbeta3/metabolism , Placentation , Progesterone/pharmacology , Animals , Estrogens/blood , Female , Fibronectins/drug effects , Immunohistochemistry , Integrin alphaVbeta3/drug effects , Pregnancy , Progesterone/blood , Reference Standards , SwineABSTRACT
Many viruses alter different stages of apoptosis of infected cells as a strategy for successful infection. Few studies have addressed mechanisms of equine herpesvirus 1 (EHV-1) strain-induced cell death. We investigated the effect of an abortigenic strain (AR8 strain) on heterologous Madin-Darby bovine kidney cells and homologous equine dermis (ED) cells cell lines. We compared morphologic and biochemical features of early and late apoptosis at different postinfection times. We investigated translocation of phosphatidylserine to the cell surface, nuclear fragmentation and changes in the cytoskeleton using flow cytometry and annexin V/propidium iodide staining, DNA laddering, terminal deoxynucleotidyl transferase UTP nick-end labeling assay and immunofluorescence staining of cytokeratin 18 cleavage. AR8 EVH-1 strain interfered with apoptosis in both cell lines, particularly during the middle stage of the replication cycle; this was more evident in ED cells. Although this antiapoptotic effect has been reported for other alpha herpesviruses, our findings may help elucidate how EHV-1 improves its infectivity during its cycle.
Subject(s)
Herpesvirus 1, Equid/pathogenicity , Virus Replication , Animals , Apoptosis , Cattle , Cells, Cultured , Flow Cytometry , Herpesvirus 1, Equid/ultrastructure , Kidney/cytology , Kidney/virology , Microscopy, Electron, TransmissionABSTRACT
Cadmium (Cd) is an industrial and environmental pollutant that produces toxic effects on gametogenesis, pre- and post-implantation embryos, and the placenta. Because the effects of acute Cd intoxication on the placenta are not well understood, we investigated changes in its glycosylated components in Cd treated dams at days 4, 7, 10 and 15 of gestation using lectin histochemistry. CdCl2 was administered to pregnant rats; control animals received sterile normal saline. Placentas were processed for DBA, Con A, SBA, PNA, UEA-I, RCA-I and WGA lectin histochemistry to evaluate changes in the carbohydrate pattern of the placenta that might modify cell interactions and contribute to embryonic alterations. Lectin binding was analyzed in the yolk sac; trophoblast giant cells; trophoblast I, II and III; spongiotrophoblast cells and endovascular trophoblast cells in the chorioallantoic placenta. Our lectin binding patterns showed that Cd caused alteration of SBA and DBA labeling of trophoblast-derived cells, which suggested increased expressions of α and ß GalNAc. Cd also caused decreased UEA-1 binding affinity, which indicated fewer α-L-Fuc residues in placentas of Cd treated dams. The nonreactivity in trophoblast I of the control placentas incubated with Con-A contrasted with the labeling in placentas of experimental dams, which indicated increased expression of terminal α-D-Man, and α-D-Glc residues. We found that Cd altered the reactivity of placenta to several lectins, which indicated modification of the glycotype presented by the fetal component of the placenta. We report that Cd exerts a deleterious effect on the glycosylation pattern of the placenta.
Subject(s)
Cadmium/toxicity , Lectins/chemistry , Placenta/drug effects , Placenta/metabolism , Animals , Female , Glycosylation , Pregnancy , Rats , Rats, Wistar , Trophoblasts/drug effects , Trophoblasts/physiologyABSTRACT
The zonary and endotheliochorial dog placenta is the most invasive placenta of carnivores. The importance of matrix metalloproteinases (MMP) in placenta invasiveness has been determined in several mammals including species with haemochorial, epitheliochorial and endotheliochorial placentation. Regarding the latter, the expression of MMP enzymes has been studied in the cat and the mature canine placenta. The aim of this study was to analyse the expression and activity of MMP-2 and MMP-9 in the early dog placenta. Placentae from 18 to 30 days of pregnancy were collected from four bitches. Two placentae from each bitch were analysed. Placental tissue from one uterine horn was fixed in formaldehyde for immunohistochemistry, while marginal haematoma, labyrinth, non-implantative and implantative endometrium from the contralateral horn were immediately frozen in dry ice for the analysis of MMP expression (Western blot [WB]) and activity (zymography). MMP-2 and MMP-9 were evidenced in the labyrinth, maternal glands and marginal haematoma; this finding was directly correlated with levels of MMP expression by WB, and with the activity of MMP-2, mainly in the haematoma (the area of major remodelling of tissues). Thus, although MMP-9 is well expressed in the early canine placenta, it is not active. Given the important role of MMPs for invasiveness, maternal-foetal angiogenesis and the establishment of a correct foetal nutrition, the results are consistent with the findings in other species in which the MMP-2 activation precedes the MMP-9 one in early placentation.
Subject(s)
Endometrium/metabolism , Matrix Metalloproteinase 2/metabolism , Matrix Metalloproteinase 9/metabolism , Placenta/metabolism , Placentation , Animals , Dogs , Female , Immunohistochemistry , PregnancyABSTRACT
This study describes the changes observed in the placentas of mice experimentally infected with an abortigenic strain of EHV-1 at mid-pregnancy and euthanized at days 3 and 4 post-infection. We analyzed microscopic vascular alterations, cell proliferation and death by immunohistochemistry, and the expression of IFN-γ, TNF-α and the IL-10 by qPCR and flow cytometry. Infected mice showed slight respiratory signs and ruffled fur during the first two days post-infection. Virus isolation and DNA detection were positive only in the lungs of the infected mice. Vascular congestion, increase in the labyrinth area, and a significant reduction in fetal capillary endothelium surface of infected placentas were found. Cell proliferation was significantly reduced in the infected placentas, whereas the apoptosis was significantly increased. IL10, TNF and IFN-γ showed different expression in the infected placentas and uteri. The effects of EHV-1 during pregnancy depend on different pathogenic mechanisms in which vascular alterations, and cell death and proliferation and local cytokine changes are compromised.