Fetal Epithelium Research Articles

Glycoconjugate-Specific Developmental Changes in the Horse Vomeronasal Organ

The vomeronasal organ (VNO) is a tubular pheromone-sensing organ in which the lumen is covered with sensory and non-sensory epithelia. This study used immunohistochemistry and lectin histochemistry techniques to evaluate developmental changes, specifically of the glycoconjugate profile, in the horse VNO epithelium. Immunostaining analysis revealed PGP9.5 expression in some vomeronasal non-sensory epithelium (VNSE) cells and in the vomeronasal receptor cells of the vomeronasal sensory epithelium (VSE) in fetuses, young foals, and adult horses. Olfactory marker protein expression was exclusively localized in receptor cells of the VSE in fetuses, young foals, and adult horses and absent in VNSE. To identify the glycoconjugate type, lectin histochemistry was performed using 21 lectins. Semi-quantitative analysis revealed that the intensities of glycoconjugates labeled with WGA, DSL, LEL, and RCA₁₂₀ were significantly higher in adult horse VSE than those in foal VSE, whereas the intensities of glycoconjugates labeled with LCA and PSA were significantly lower in adult horse VSE. The intensities of glycoconjugates labeled with s-WGA, WGA, BSL-II, DSL, LEL, STL, ConA, LCA, PSA, DBA, SBA, SJA, RCA₁₂₀, jacalin, and ECL were significantly higher in adult horse VNSE than those in foal VNSE, whereas the intensity of glycoconjugates labeled with UEA-I was lower in adult horse VNSE. Histochemical analysis of each lectin revealed that various glycoconjugates in the VSE were present in the receptor, supporting, and basal cells of foals and adult horses. A similar pattern of lectin histochemistry was also observed in the VNSE of foals and adult horses. In conclusion, these results suggest that there is an increase in the level of N-acetylglucosamine (labeled by WGA, DSL, LEL) and galactose (labeled by RCA₁₂₀) in horse VSE during postnatal development, implying that they may influence the function of VNO in adult horses.

Effect of porcine reproductive and respiratory syndrome virus 2 on angiogenesis and cell proliferation at the maternal-fetal interface.

Angiogenesis and cell proliferation in reproductive tissues are essential events for the maintenance of pregnancy, and alterations can lead to compromised fetal development and survival. Porcine reproductive and respiratory syndrome virus 2 (PRRSV-2) induces reproductive disease with negative financial and production impact on the swine industry. PRRSV-2 infection alters placental physiology through inflammatory and apoptotic pathways, yet fetal susceptibility varies. This study aimed to evaluate angiogenesis and cell proliferation in the porcine maternal-fetal interface (MFI) and determine if these physiological processes were altered by PRRSV-2 infection. Thirty-one pregnant gilts were inoculated with PRRSV-2 at gestation day 86 ± 0.4 (mean ± SD). Seven control gilts were sham-inoculated. All gilts were euthanized at 12 days postinoculation. Angiogenesis and cell proliferation were determined through the detection of vascular endothelial growth factor (VEGF) and Ki-67, respectively, using immunofluorescence of the MFI from 4 fetal resilience groups: uninfected (UNIF), high viral load–viable (HVL-VIA), and HVL-meconium-stained (MEC) from PRRSV-infected gilts, as well from sham-inoculated (CON) gilts. VEGF immunolabeling in the uterine submucosa was significantly lower in MEC compared with UNIF and HVL-VIA groups. Significantly greater Ki67 immunolabeling was detected in the trophoblasts of CON fetuses versus all other groups, and in uterine epithelium of CON and UNIF fetuses versus HVL-VIA and MEC. These results suggest that fetal resilience may be related to greater cell proliferation in uterine epithelium, and fetal compromise with reduced uterine submucosal angiogenesis, except fetuses with intrauterine growth restriction, in which inherently lower submucosal angiogenesis may be protective against PRRSV infection.

Differential H. pylori-Induced MAPK Responses Regulate Lewis Antigen Expression and Colonization Density on Gastric Epithelial Cells Between Children and Adults.

Helicobacter pylori causes gastrointestinal diseases, the manifestations of diseases are more serious in adults than in children. Lewis antigen expressions on the gastric epithelium serves as receptors targeted by H. pylori. Moreover, the MAPK signaling pathway involves glycoprotein synthesis of Lewis antigens. We aimed to investigate whether differences in H. pylori-induced MAPK responses mediate gastric Lewis antigens expression and colonization density differently in children and adults. We used human stomach fetal epithelium (HSFE) and SV40-immortalized human normal gastric epithelial (GES-1) cell lines to mimic primary gastric epithelium of children and adults, respectively. H. pylori colonization intensity and Lewis antigens were significantly higher in GES-1 than in HSFE cells, whereas IL-8 and IL-6 levels were significantly higher in HSFE than in GES-1 cells after infection. c-Jun N-terminal kinase (JNK) siRNA and inhibitor (SP600125) experiments showed that Lewis antigen expression and H. pylori colonization were reduced in GES-1 cells but increased in HSFE cells. Furthermore, p-p38 intensity was significantly higher in the superficial epithelium of the children than in the adults with/without H. pylori infection. The overexpression of p38 in GES-1 cells downregulated H. pylori-induced JNK activity mimicking H. pylori infection in children. In conclusion, a higher p38 expression in gastric epithelium counteracting JNK activity in children may contribute to lower Lewis antigen expression and colonization density than in adults after H. pylori infection.

Distinct properties of pure- and mixed-type high-grade fetal lung adenocarcinomas by genetic profiling and transcription factor expression

The clinicopathological differences among high-grade fetal lung adenocarcinomas completely comprising tumor cells that resemble fetal lung epithelium (pure type) and those with fetal lung-like components admixed with conventional adenocarcinoma cells (mixed type) remain undetermined. Here, we examined the clinicopathological, immunohistochemical, and molecular features of 11 lung adenocarcinomas with fetal lung-like morphology among 3895 consecutive cases of primary lung cancer based on the expression pattern of transcription factors. According to the current WHO classification, two cases (0.05%) were categorized as low-grade fetal adenocarcinoma, two cases (0.05%) were pure-type high-grade fetal adenocarcinoma, five cases (0.1%) were mixed-type high-grade fetal adenocarcinoma, and the remaining two cases (0.05%) were lung adenocarcinoma with high-grade fetal features (fetal lung-like morphology occupied less than 50%). CTNNB1 mutations were exclusively identified in low-grade fetal adenocarcinomas. In contrast, mixed-type high-grade fetal adenocarcinoma or lung adenocarcinoma with high-grade fetal features frequently harbored mitogenic drivers including EGFR mutations. Furthermore, almost all tumor cells expressed CDX2 and HNF4α in both cases of pure-type high-grade fetal lung adenocarcinoma, but lacked TTF-1 positivity. In contrast, TTF-1 was frequently expressed in mixed-type high-grade fetal lung adenocarcinoma and in lung adenocarcinoma with high-grade fetal features. Our data suggest similar prevalence of low-grade fetal lung adenocarcinoma and pure-type high-grade fetal lung adenocarcinoma, and indicate that pure- and mixed-type high-grade fetal lung adenocarcinomas are distinct, with the former akin to low-grade fetal adenocarcinoma with respect to purely embryonic morphology and absence of common lung adenocarcinoma mitogenic drivers, and the latter being genetically and transcriptionally related to conventional lung adenocarcinoma.

Intestinal epithelial tuft cell induction is negated by a murine helminth and its secreted products.

Helminth parasites are adept manipulators of the immune system, using multiple strategies to evade the host type 2 response. In the intestinal niche, the epithelium is crucial for initiating type 2 immunity via tuft cells, which together with goblet cells expand dramatically in response to the type 2 cytokines IL-4 and IL-13. However, it is not known whether helminths modulate these epithelial cell populations. In vitro, using small intestinal organoids, we found that excretory/secretory products (HpES) from Heligmosomoides polygyrus blocked the effects of IL-4/13, inhibiting tuft and goblet cell gene expression and expansion, and inducing spheroid growth characteristic of fetal epithelium and homeostatic repair. Similar outcomes were seen in organoids exposed to parasite larvae. In vivo, H. polygyrus infection inhibited tuft cell responses to heterologous Nippostrongylus brasiliensis infection or succinate, and HpES also reduced succinate-stimulated tuft cell expansion. Our results demonstrate that helminth parasites reshape their intestinal environment in a novel strategy for undermining the host protective response.

Decreased tight junction protein intensity in the placenta of porcine reproductive and respiratory syndrome virus-2 infected fetuses

IntroductionExisting strategies to control porcine reproductive and respiratory syndrome (PRRS) are not completely effective and require alternative approaches. Although intrauterine growth restricted (IUGR) fetuses are more resilient to transplacental PRRS virus-2 (PRRSV2) infection compared to normal fetuses, the exact mechanisms are unknown. The objective of this research was to assess abundance and localization of a subset of tight junction (TJ) proteins in the maternal-fetal interface and any alterations that may affect the movement of nutrients or PRRSV2 across the epitheliochorial placenta. MethodsParaffin-embedded samples of placenta from non-infected control (CTRL) and PRRSV2 infected fetuses (IUGR, non(N)-IUGR, meconium-stained (MEC) (n = 6 per group) were randomly selected from a large challenge trial and immunostained for claudins (CLDN) 1, 3, 4, 7 and tight junction protein 1 (TJP1). Immunostaining intensity was semi-subjectively scored by region. ResultsIntensity of CLDN1 was lower in placenta of IUGR, MEC, and N-IUGR fetuses compared to CTRL, mainly in fetal epithelium and maternal endothelial cells (MECL). CLDN4 intensity was lower in MECL of IUGR compared to CTRL and MEC fetuses. TJP1 intensity was lower in maternal and fetal epithelia of placenta within IUGR, MEC, and N-IUGR fetuses versus CTRL. DiscussionDifferences were mainly observed between PRRSV2 infected and non-infected groups indicating TJ integrity was affected by PRRSV2 infection. These results provide insights into the potential mechanisms of transplacental transmission of PRRSV2; however, since only CLDN4 differed amongst the infected groups, PRRSV2 induced changes in TJ integrity do not appear to explain variation in fetal outcomes after infection.

PSVIII-37 Late-Breaking Abstract: Effects of feeding a vitamin and mineral supplement and (or) an energy supplement on the abundance of SLC7A5 transporter in beef heifer placentomes at d 83 of gestation

Abstract Objectives were to evaluate effects of vitamin and mineral (VTM) and (or) energy (NRG) supplements on the abundance of the neutral amino acid (AA) transporter SLC7A5 in heifer placentomes at d 83 of gestation. Thirty-five crossbred Angus heifers (Body weight = 359.5 ± 7.1 kg) were randomly assigned to one of four treatments (n = 8 or 9/group) in a 2 × 2 factorial arrangement with main effects of vitamin and mineral (NoVTM or VTM) and energy (NoNRG or NRG) supplements. The VTM treatment was initiated 71 to 148 d before artificial insemination (AI). The NRG treatment was initiated at AI. Heifers were ovariohysterectomized at d 83 of gestation. Placentome cross sections (5 μm) were mounted onto glass slides, deparaffinized, and stained for SLC7A5. Fluorescent images of tissue areas including fetal membranes, caruncles (CAR), endometrial epithelium (EP), superficial glands (SG), deep glands (DG), and myometrium were obtained and analyzed for relative intensity of fluorescence within the areas of interest as an indicator of transporter abundance. The abundance of SLC7A5 was influenced by a VTM × NRG interaction (P = 0.05) in DG, with NoVTM-NRG being greater than heifers receiving VTM. Abundance of SLC7A5 was not influenced by NRG (P > 0.12) in any structures evaluated. In EP and SG abundance of SLC7A5 was less (P ≤ 0.05) in VTM than NoVTM heifers and tended to be less in CAR (P = 0.07). Our previous work revealed no differences in maternal serum AA concentrations, but VTM heifers had greater AA concentrations in allantoic fluid than NoVTM. We hypothesize that transporter efficiency may be reduced in NoVTM heifers, with greater transporter abundance being a mechanism of compensation to supply AA to the fetus. Together, these data help to further elucidate mechanisms of AA uptake and balance between the maternal and fetal compartments.

Lipid metabolism is altered in maternal, placental, and fetal tissues of ewes with small for gestational age fetuses†.

Nutrient restriction (NR) has the potential to negatively impact birthweight, an indicator of neonatal survival and lifelong health. Those fetuses are termed as small for gestational age (SGA). Interestingly, there is a spectral phenotype of fetal growth rates in response to NR associated with changes in placental development, nutrient and waste transport, and lipid metabolism. A sheep model with a maternal diet, starting at Day 35, of 100% National Research Council (NRC) nutrient requirements (n=8) or 50% NRC (n=28) was used to assess alterations in fetuses designated NR SGA (n=7) or NR NonSGA (n=7) based on fetal weight at Day 135 of pregnancy. Allantoic fluid concentrations of triglycerides were greater in NR SGA fetuses than 100% NRC and NR NonSGA fetuses at Day 70 (P<0.05). There was a negative correlation between allantoic fluid concentrations of triglycerides (R2=0.207) and bile acids (R2=0.179) on Day 70 and fetal weight at Day 135 for NR ewes (P<0.05). Bile acids were more abundant in maternal and fetal blood for NR SGA compared to 100% NRC and NR NonSGA ewes (P<0.05). Maternal blood concentrations of NEFAs increased in late pregnancy in NR NonSGA compared to NR SGA ewes (P<0.05). Protein expression of fatty acid transporter SLC27A6 localized to placentomal maternal and fetal epithelia and decreased in Day 70 NR SGA compared to 100% NRC and NR NonSGA placentomes (P<0.05). These results identify novel factors associated with an ability of placentae and fetuses in NR NonSGA ewes to adapt to, and overcome, nutritional hardship during pregnancy.

Parasitic helminths induce fetal-like reversion in the intestinal stem cell niche.

Epithelial surfaces form critical barriers to the outside world and are continuously renewed by adult stem cells1. Whereas epithelial stem cell dynamics during homeostasis are increasingly well understood, how stem cells are redirected from a tissue-maintenance program to initiate repair after injury remains unclear. Here, we examined infection by Heligmosomoides polygyrus (Hp), a co-evolved pathosymbiont of mice, to assess the epithelial response to disruption of the mucosal barrier. Hp disrupts tissue integrity by penetrating the duodenal mucosa, where it develops while surrounded by a multicellular granulomatous infiltrate2. Unexpectedly, intestinal stem cell (ISC) markers, including Lgr53, were lost in crypts overlying larvae-associated granulomas, despite continued epithelial proliferation. Granuloma-associated Lgr5− crypt epithelia activated an interferon-gamma (IFNγ)-dependent transcriptional program, highlighted by Sca-1 expression, and IFNγ-producing immune cells were found in granulomas. A similar epithelial response accompanied systemic activation of immune cells, intestinal irradiation, or ablation of Lgr5+ ISCs. Granuloma-associated crypt cells generated fetal-like spheroids in culture, and a sub-population of Hp-induced cells activated a fetal-like transcriptional program, demonstrating that adult intestinal tissues can repurpose aspects of fetal development. Thus, re-initiation of the developmental program represents a fundamental mechanism by which the intestinal crypt can remodel to sustain function after injury.

SINONASAL TERATOCARCINOSARCOMA - A RARE CASE REPORT

Sinonasal teratocarcinosarcoma is a rare and highly malignant tumor characterized by complex admixture of teratomatous and carcinosarcomatous component. Here, we report a case of 72-year-old male presented with complaints of epistaxis, recurrent sinusitis, nasal stuffiness, and nasal blockade for past 5-6 months and his computed tomography report revealed as an inflammatory sinonasal disease with polyposis. Histopathology examination showed fragments of upper respiratory mucosa admixed with fragments of tumor composed of benign and malignant counterparts of both epithelial and mesenchymal components in a variety of histologic patterns. Also noted were the characteristic squamous epithelium with “fetal-type” clear cells, areas of necrosis, mitotic figure and sheets of undifferentiated blastemal like cells. Immunohistochemistry demonstrated positive staining for cytokeratin, epithelial membrane antigen, vimentin, synaptophysin, S-100, chromogranin, and CD99. This polyimmunophenotypic characteristic and diverse histological differentiation provided the basis for the diagnosis. Knowing its highly aggressive behavior, the patient should be treated aggressively and followed up closely. Key words: Carcinosarcom, Fetal epithelium, Polyimmunophenotypic, Teratoid

Transient Tachypnea of the Newborn

* Abbreviations: ABG, : arterial blood gas ALPS, : Antenatal Late Preterm Steroids CBC, : complete blood cell CPAP, : continuous positive airway pressure CRP, : C-reactive protein ENaC, : epithelial sodium channel FiO2, : fraction of inspired oxygen IV, : intravenous TTN, : transient tachypnea of the newborn It can be challenging to diagnose and provide optimal treatment for transient tachypnea of the newborn. After completing this article, readers should be able to: 1. Understand the pathophysiology of transient tachypnea of the newborn (TTN). 2. Identify risk factors, clinical symptoms, and radiographic findings in infants with TTN. 3. Appreciate the differential diagnoses for TTN. 4. Describe the typical clinical course of an infant with TTN. Fetal lungs are filled with liquid that is crucial for normal lung growth. After birth, this fetal alveolar fluid is cleared through pulmonary epithelial, vascular, and lymphatic channels. A delay in fluid clearance leads to ineffective gas exchange and results in respiratory distress. This impermanent condition of retained fetal lung fluid is known as transient tachypnea of the newborn (TTN). (1)(2) TTN is one of the leading causes of neonatal respiratory distress and is therefore an important diagnosis to consider, identify correctly, and manage. (3) It is generally a benign, self-limited condition that presents shortly after birth and occurs in infants of any gestational age. Diagnosis of TTN is based on an infant’s clinical presentation, physical examination findings, and classic chest radiographic findings. The management consists of supportive care, with symptoms generally resolving by 24 to 72 hours of age. As early as 6 weeks of gestation, the fetal lung epithelium begins to secrete alveolar fluid at 2 mL/kg per hour, increasing to a rate of 5 mL/kg per hour at term. (4) This liquid is crucial for normal lung growth and contributes to the volume of amniotic fluid that surrounds the fetus. A few days before the onset of spontaneous vaginal delivery, fluid production decreases. (5) With the onset of labor, maternal hormones such as epinephrine and glucocorticoids stimulate the fetal lungs to begin absorption of alveolar fluid through activation of an amiloride-sensitive …

Follistatin-like 1 expression is decreased in the alveolar epithelium of hypoplastic rat lungs with nitrofen-induced congenital diaphragmatic hernia

Background/purposePulmonary hypoplasia (PH), characterized by incomplete alveolar development, remains a major therapeutic challenge associated with congenital diaphragmatic hernia (CDH). Follistatin-like 1 (Fstl1) is a crucial regulator of alveolar formation and maturation, which is strongly expressed in distal airway epithelium. Fstl1-deficient mice exhibit reduced airspaces, impaired alveolar epithelial cell differentiation, and insufficient production of surfactant proteins similar to PH in human CDH. We hypothesized that pulmonary Fstl1 expression is decreased during alveolarization in the nitrofen-induced CDH model. MethodsTimed-pregnant rats received nitrofen or vehicle on gestational day 9 (D9). Fetal lungs were harvested on D18 and D21 and divided into control−/nitrofen-exposed specimens. Alveolarization was assessed using morphometric analysis techniques. Pulmonary gene expression of Fstl1 was determined by qRT-PCR. Immunofluorescence-double-staining for Fstl1 and alveolar epithelial marker surfactant protein C (SP-C) was performed to evaluate protein expression/localization. ResultsRadial alveolar count was significantly reduced in hypoplastic lungs of nitrofen-exposed fetuses with significant down regulation of Fstl1 mRNA expression on D18 and D21 compared to controls. Confocal-laser-scanning-microscopy revealed strikingly diminished Fstl1 immunofluorescence and SP-C expression in distal alveolar epithelium of nitrofen-exposed fetuses with CDH-associated PH on D18 and D21 compared to controls. ConclusionsDecreased expression of Fstl1 in alveolar epithelium may disrupt alveolarization and pulmonary surfactant production, thus contributing to the development of PH in the nitrofen-induced CDH model. Level of evidence2b (Centre for Evidence-Based Medicine, Oxford).

Cochlear epithelial of dog fetuses: a new source of multipotent stem cells.

Hearing loss caused by the damage of cochlea sensory cells or neurons is a common human disease, but also affects dogs and other animals. To test their progenitor nature as potential value for future therapies, we characterized cells derived from the cochlear epithelium in dog fetuses. In total, 8 fetuses of 35-40days of gestation, derived from castration campaigns, were investigated. Cells were analysed by the MTT colorimetric assay and in regard to cell cycle, differentiation capacities, immunophenotypes and qPCR analysis. In culture, cells had a fibroblast-like morphology. Phenotypic immunocharacterization showed positive staining for mesenchymal stem cell and pluripotency markers and were negative for hematopoietic cell markers. Cells possessed differentiation capacity for the three main cell lineages: osteogenic, adipogenic and chondrogenic, altogether indicating their nature as mesenchymal stem cells. Thus, cells derived from fetal cochlear tissues indeed may provide valuable sources of progenitor cells for cell therapy of canine deafness and other diseases.

Proteomic profiling of fetal esophageal epithelium, esophageal cancer, and tumor-adjacent esophageal epithelium and immunohistochemical characterization of a representative differential protein, PRX6.

AIMTo understand the molecular mechanism of esophageal cancer development and provide molecular markers for screening high-risk populations and early diagnosis.METHODSTwo-dimensional electrophoresis combined with mass spectrometry were adopted to screen differentially expressed proteins in nine cases of fetal esophageal epithelium, eight cases of esophageal cancer, and eight cases of tumor-adjacent normal esophageal epithelium collected from fetuses of different gestational age, or esophageal cancer patients from a high-risk area of esophageal cancer in China. Immunohistochemistry (avidin-biotin-horseradish peroxidase complex method) was used to detect the expression of peroxiredoxin (PRX)6 in 91 cases of esophageal cancer, tumor-adjacent normal esophageal tissue, basal cell hyperplasia, dysplasia, and carcinoma in situ, as well as 65 cases of esophageal epithelium from fetuses at a gestational age of 3-9 mo.RESULTSAfter peptide mass fingerprint analysis and search of protein databases, 21 differential proteins were identified; some of which represent a protein isoform. Varying degrees of expression of PRX6 protein, which was localized mainly in the cytoplasm, were detected in adult and fetal normal esophageal tissues, precancerous lesions, and esophageal cancer. With the progression of esophageal lesions, PRX6 protein expression showed a declining trend (P < 0.05). In fetal epithelium from fetuses at gestational age 3-6 mo, PRX6 protein expression showed a declining trend with age (P < 0.05). PRX6 protein expression was significantly higher in well-differentiated esophageal cancer tissues than in poorly differentiated esophageal cancer tissues (P < 0.05).CONCLUSIONDevelopment and progression of esophageal cancer result from interactions of genetic changes (accumulation or superposition). PRX6 protein is associated with fetal esophageal development and cancer differentiation.

Signal molecules as biomarkers of prediction of the premature rupture of membranes (clinicodiagnostic aspects)

One of the most commonly encountered pregnancy complications is the premature rupture of membranes. This pathology results in the increase of frequency of operative delivery, birth traumatism and neonatal complications. The purpose of the research described was verification of key signal molecules, providing integrity of fetal membranes, with subsequent development of possible biomarkers of non-invasive prediction of the premature rupture of membranes. This work presents the comparison studies of expression of VEGF, MMP-9, connexin 37, connexin 40, endorphins, enkephalins, actin, miosin in a buccal epithelium and fetal membranes for 70 patients of the basic group (with premature rupture of membranes) and for 70 patients of the control group (with timely rupture of membranes). Research of fetal membranes and buccal epithelium was carried out by means of primary monoclonal mouse antibodies to the investigated markers. The universal basis set was used as the secondary antibodies, containing of biotinylated anti-mouse immunoproteins. The study of the preparations was carried out in the confocal microscope OLYMPUS FLUOVIEW FV 1000 at the image enlargement of ×400 and ×1000 with use of the system MRC-1024, with the software suite for computer processing OLYMPUS FLUOVIEW 5.0. Statistical processing of the material was carried out with the application of the standard statistical software suite Statistica 10.0. Obtained in a group with the premature rupture of membranes and in a control group were the reliable differences of expression of MMP-9, VEGF, connexin 37 and connexin 40 were. The multifactorial analysis of the indices of expression of signal molecules allowed to discover the high information significance for premature rupture of membranes prediction, matrix metalloproteinase ММP-9, connexin 37 and connexin 40, as well as VEGF. Matrix metalloproteinase ММP-9, connexin 37 and connexin 40, VEGF can be considered as non-invasive markers of premature rupture of membranes prediction.

CYP21A2 expression is localized in the developing distal epithelium of the human perinatal lung and is compatible with in situ production and intracrine actions of active glucocorticoids

Glucocorticoids play essential roles in lung development. We investigated for expression of CYP21A2 (21-hydroxylase) as well as for the presence of the corresponding protein and identification of CYP21A2-expressing cells in several human developing lungs. Expression of some related genes was also assessed. CYP21A2 and CYP17A1 (P450c17) mRNAs were found in all the 34 lung samples from 17 to 40 weeks’ gestation at variable levels. No correlation was found according to sex but a correlation with age was detected for CYP17A1 only. In contrast, CYP11B1 (11β-hydroxylase)- and CYP11B2 (aldosterone synthase)-mRNAs were not detected. Significant levels of the CYP21A2 protein were detected in all the analyzed samples, while only very low signals were detected for CYP17A1 protein. In situ hybridization revealed that CYP21A2 was almost exclusively expressed in the distal epithelium. It was reported that the lung distal epithelium of human fetuses also express 11β-hydroxysteroid dehydrogenase type 2, which catalyzes cortisol inactivation into cortisone. Based on this information, intracrine glucocorticoid actions should take place from CYP21A2 products through the glucocorticoid receptor in the absence of cortisol. In contrast, mineralocorticoid receptor activation did not seem to depend on deoxycorticosterone produced from local activity of CYP21A2 because of the reported circulating amounts of aldosterone.

Decreased expression of monocarboxylate transporter 1 and 4 in the branching airway epithelium of nitrofen-induced congenital diaphragmatic hernia

Background/PurposeMonocarboxylate transporters (MCTs) are crucial for the maintenance of intracellular pH homeostasis in developing fetal lungs. MCT1/4 is strongly expressed by epithelial airway cells throughout lung branching morphogenesis. Functional inhibition of MCT1/4 in fetal rat lung explants has been shown to result in airway defects similar to pulmonary hypoplasia (PH) in congenital diaphragmatic hernia (CDH). We hypothesized that pulmonary expression of MCT1/4 is decreased during lung branching morphogenesis in the nitrofen model of CDH-associated PH. MethodsTimed-pregnant rats received nitrofen or vehicle on gestational day 9 (D9). Fetuses were harvested on D15, D18, and D21, and divided into control and nitrofen-exposed group. Pulmonary gene expression levels of MCT1/4 were analyzed by qRT-PCR. Immunofluorescence staining for MCT1/4 was combined with E-cadherin in order to evaluate protein expression in branching airway tissue. ResultsRelative mRNA levels of MCT1/4 were significantly reduced in lungs of nitrofen-exposed fetuses on D15, D18, and D21 compared to controls. Confocal laser scanning microscopy confirmed markedly decreased immunofluorescence of MCT1/4 in distal bronchial and primitive alveolar epithelium of nitrofen-exposed fetuses on D15, D18, and D21 compared to controls. ConclusionDecreased expression of MCT1/4 in distal airway epithelium may disrupt lung branching morphogenesis and thus contribute to the development of PH in the nitrofen-induced CDH model.

Trop2 marks transient gastric fetal epithelium and adult regenerating cells after epithelial damage.

ABSTRACTMouse fetal intestinal progenitors lining the epithelium prior to villogenesis grow as spheroids when cultured ex vivo and express the transmembrane glycoprotein Trop2 as a marker. Here, we report the characterization of Trop2-expressing cells from fetal pre-glandular stomach, growing as immortal undifferentiated spheroids, and their relationship with gastric development and regeneration. Trop2+ cells generating gastric spheroids differed from adult glandular Lgr5+ stem cells, but appeared highly related to fetal intestinal spheroids. Although they shared a common spheroid signature, intestinal and gastric fetal spheroid-generating cells expressed organ-specific transcription factors and were committed to intestinal and glandular gastric differentiation, respectively. Trop2 expression was transient during glandular stomach development, being lost at the onset of gland formation, whereas it persisted in the squamous forestomach. Undetectable under homeostasis, Trop2 was strongly re-expressed in glands after acute Lgr5+ stem cell ablation or following indomethacin-induced injury. These highly proliferative reactive adult Trop2+ cells exhibited a transcriptome displaying similarity with that of gastric embryonic Trop2+ cells, suggesting that epithelium regeneration in adult stomach glands involves the partial re-expression of a fetal genetic program.

Characterization of mesenchymal cells beneath cornification of the fetal epithelium and epidermis at the face: an immunohistochemical study using human fetal specimens.

Fetal development of the face involves a specific type of cornification in which keratinocytes provide a mass or plug to fill a cavity. The epithelial-mesenchymal interaction was likely to be different from that in the usual skin. We examined expression of intermediate filaments and other mesenchymal markers beneath cornification in the fetal face. Using sections from 5 mid-term human fetuses at 14–16 weeks, immunohistochemistry was conducted for cytokeratins (CK), vimentin, nestin, glial fibrilary acidic protein, desmin, CD34, CD68 and proliferating cell nuclear antigen (PCNA). Fetal zygomatic skin was composed of a thin stratum corneum and a stratum basale (CK5/6+, CK14+, and CK19+) and, as the intermediate layer, 2–3 layered large keratinocytes with nucleus. The basal layer was lined by mono-layered mesenchymal cells (CD34+ and nestin+). Some of basal cells were PCNA-positive. In the keratinocyte plug at the external ear and nose, most cell nuclei expressed PCNA, CK5/6, CK14, and CK19. Vimentin-positive mesenchymal cells migrated into the plug. The PCNA-positive nucleus as well as mesenchymal cell migration was not seen in the lip margin in spite of the thick keratinocyte layer. The lingual epithelium were characterized by the CK7-positive stratum corneum as well as the thick mesenchymal papilla. CD68-positive macrophages were absent in the epidermis/epithelium. Being different from usual cornification of the skin, loss of a mesenchymal monolayer as well as superficial migration of mesenchymal cells might connect with a specific differentiation of keratinocyte to provide a plug at the fetal nose and ear.

26 The Cellular Plasticity of Fetal and Adult Intestinal Epithelium and Its Functional Evaluation by Grafting Assay

histology), circularity values increased up to 0.9. C57BL/6 enteroids showed a dose dependent increase in circularity in response to TNF (10-1000ng/ml) or IFN γ (1-100ng/ml) after 24 and 48 hours. Circularity did not however increase in Nfkb2-/enteroids to the same extent following these stimuli. C57BL/6 BMDCs or their CM or LPS alone did not induce an increase in the circularity of C57BL/6, Nfkb2 or Nfkb1-/enteroids. However, when stimulated with 1μg/ml LPS, C57BL/6 BMDCs and their CM induced a significant >2-fold increase in the circularity of C57BL/6 and Nfkb1-/enteroids. Nfkb2 derived enteroids showed no such increase. LPS stimulated BMDCs from Nfkb2-/mice were able to induce a significant increase in C57BL/6 enteroid circularity of a similar magnitude to C57BL/6 BMDCs. Conclusions:Circularity correlates well with cell death and shedding in murine enteroid culture. Deletion of Nfkb2 from intestinal epithelial cells prevents epithelial cell destruction in response to secreted immune cell factors. Therapeutics that specifically target NFKB2 signalling in intestinal epithelial cells may therefore prevent loss of intestinal barrier function and thus ameliorate intestinal inflammation.