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GATAe-dependent and -independent expressions of genes in the differentiated endodermal midgut of Drosophila.

Two sequentially-expressed GATA factor genes, serpent (srp) and GATAe, are essential for development of the Drosophila endoderm. The earliest endodermal GATA gene, srp, has been thought to specify the endodermal fate, activating the second GATA gene GATAe, and the latter continues to be expressed in the endodermal midgut throughout life. Previously, we proposed that GATAe establishes and maintains the state of terminal differentiation of the midgut, since some functional genes in the midgut require GATAe activity for their expression. To obtain further evidence of the role of GATAe, we searched for additional genes that are expressed specifically in the midgut in late stages, and examined responses of a total of selected 15 genes to the depletion and overexpression of GATAe. Ten of the 15 genes failed to be expressed in the embryo deficient for GATAe activity, but, the other five genes did not require GATAe. Instead, srp is required for activating the five genes. These observations indicate that GATAe activates a major subset of genes in the midgut, and some other pathway(s) downstream of srp activates other genes.

Animals↗

Disturbing endoderm signaling to anterior neural plate of vertebrates by the teratogen cadmium.

Cadmium accumulation in the mouse gut endoderm occurs until the closure of the vitelline duct (day 9 post-coitus; p.c.), producing anterior neural tube defects (NTD). The anterior part of the primitive endoderm, designated as the primary signaling center for anterior patterning, expresses several transcription factors of importance for head formation. Here, we studied the expression levels of some of these transcription factors (Hesx1, HNF3beta, Cerl, Otx2 and Sox2), and cell death induced after single cadmium administration to dams on days 7, 8 and 9 p.c. Stage specific down-regulation of Hesx1, Cerl, and Sox2, and an up-regulation of HNF3beta were observed. No effect was seen in Otx2 expression levels. Cell death was increased in the neuroepithelium of the cranial neural folds, and in areas where neural crest cells migrate, but not in the gut endoderm. It is proposed that cadmium-induced NTD is due to interference with head-inductive signals from the endoderm to the adjacent layers.

Animals↗

BMP receptor IA is required in the mammalian embryo for endodermal morphogenesis and ectodermal patterning.

BMPRIA is a receptor for bone morphogenetic proteins with high affinity for BMP2 and BMP4. Mouse embryos lacking Bmpr1a fail to gastrulate, complicating studies on the requirements for BMP signaling in germ layer development. Recent work shows that BMP4 produced in extraembryonic tissues initiates gastrulation. Here we use a conditional allele of Bmpr1a to remove BMPRIA only in the epiblast, which gives rise to all embryonic tissues. Resulting embryos are mosaics composed primarily of cells homozygous null for Bmpr1a, interspersed with heterozygous cells. Although mesoderm and endoderm do not form in Bmpr1a null embryos, these tissues are present in the mosaics and are populated with mutant cells. Thus, BMPRIA signaling in the epiblast does not restrict cells to or from any of the germ layers. Cells lacking Bmpr1a also contribute to surface ectoderm; however, from the hindbrain forward, little surface ectoderm forms and the forebrain is enlarged and convoluted. Prechordal plate, early definitive endoderm, and anterior visceral endoderm appear to be expanded, likely due to defective morphogenesis. These data suggest that the enlarged forebrain is caused in part by increased exposure of the ectoderm to signaling sources that promote anterior neural fate. Our results reveal critical roles for BMP signaling in endodermal morphogenesis and ectodermal patterning.

Animals↗

Parietal endoderm secreted SPARC promotes early cardiomyogenesis in vitro.

Cardiomyogenesis proceeds in the presence of signals emanating from extra-embryonic lineages emerging before and during early eutherian gastrulation. In embryonic stem cell derived embryoid bodies, primitive endoderm gives rise to visceral and parietal endoderm. Parietal endoderm undergoes an epithelial to mesenchymal transition shortly before first cardiomyocytes start to contract rhythmically. Here, we demonstrate that Secreted Protein, Acidic, Rich in Cysteine, SPARC, predominantly secreted by mesenchymal parietal endoderm specifically promotes early myocardial cell differentiation in embryoid bodies. SPARC enhanced the expression of bmp2 and nkx2.5 in embryoid bodies and fetal cardiomyocytes. Inhibition of either SPARC or Bmp2 attenuated in both cases cardiomyogenesis and downregulated nkx2.5 expression. Thus, SPARC directly affects cardiomyogenesis, modulates Bmp2 signaling, and contributes to a positive autoregulatory loop of Bmp2 and Nkx2.5 in cardiomyocytes.

Animals↗

VegT induces endoderm by a self-limiting mechanism and by changing the competence of cells to respond to TGF-beta signals.

The maternal determinant VegT is required for both endoderm and mesoderm formation by the Xenopus embryo. An important downstream mediator of VegT action is Xsox17, which has been proposed to be induced in cell-autonomous, then signal-dependent phases. We show that Xsox17 is a direct VegT target, but that direct induction of Xsox17 by VegT is rapidly inhibited. This inhibition is relieved by TGF- beta signalling, to which the future endoderm cell is sensitised by VegT, resulting in the observed dependence on cell contact for maintained Xsox17 expression. We propose that this change in regulation is a consequence of a VegT-induced repressor, inhibiting direct induction of early endoderm markers by VegT, and contributing to the formation of the boundary of the endodermal domain.

Animals↗

Interplay of negative and positive signals controls endoderm-specific expression of the ascidian Cititf1 gene promoter.

Cititf1 is an early and specific marker of endoderm development in Ciona intestinalis [ Development 126, 5149]. Here, we examine Cititf1 transcriptional regulation focusing in particular on its endodermal restricted expression. Through the analysis of Ciona embryos, electroporated with different portions of Cititf1 5'-flanking region fused to lacZ, we characterized a minimal 300-bp cis-regulatory sequence able to closely reproduce the spatial and temporal expression pattern of the endogenous gene. This enhancer contains at least three distinct regulatory regions, two of which are responsible for activation of transcription in the endoderm and in the mesenchyme, respectively, while the third is a negative control element that represses mesenchyme transcription. We have further defined the sequences responsible for transcriptional activation in the endoderm by clustered point mutations and DNA-binding assays.

Animals↗

Initiation and early patterning of the endoderm.

We review the early stages of endoderm formation in the major animal models. In Amphibia maternal molecules are important in initiating endoderm formation. This is followed by successive signaling events that establish and then pattern the endoderm. In other organisms there are differences in endodermal development, particularly in the initial, prephylotypic stages. Later many of the same key families of transcription factors and signaling cassettes are used in all animals, but more work will be needed to establish exact evolutionary homologies.

Amphibians↗

Endoderm development: from patterning to organogenesis.

Although the ectoderm and mesoderm have been the focus of intensive work in the recent era of studies on the molecular control of vertebrate development, the endoderm has received less attention. Because signaling must occur between germ layers in order to achieve a properly organized body, our understanding of the coordinated development of all organs requires a more thorough consideration of the endoderm and its derivatives. This review focuses on present knowledge and perspectives concerning endoderm patterning and organogenesis. Some of the classical embryology of the endoderm is discussed and the progress and deficiencies in cellular and molecular studies are noted.

Animals↗

Rat yolk sac explants as a system for studying the regulation of endodermal genes: down-regulation of the alpha-fetoprotein gene by dexamethasone and phorbol ester.

The visceral yolk sac is a fetal membrane with essential placental functions. It is the major site of synthesis of alpha-fetoprotein (AFP), the most abundant plasma protein in the fetus. We developed a system of rat yolk sac explants in serum-free culture medium to study the regulation of endodermal gene expression in yolk sac. The explanted yolk sac tissues retained their double-sided morphology for up to 48 hours. The epithelial cells of both layers remained tightly joined on a basement membrane as seen by light and electron microscopy. This probably accounts for the continued expression of several endodermal cell-specific markers. The levels of mRNA encoding AFP, vitamin D-binding protein (DBP), hepatocyte nuclear factor 1alpha and beta transcription factors did not change during the 48-hour culture period. This reflects the stability of the differentiation state of the yolk sac endodermal cells. Dexamethasone and phorbol ester (TPA) specifically reduced the AFP mRNA level without affecting that of DBP. This suggests that these transduction pathways are functional in the yolk sac during this period of gestation and could be involved in the physiological down-regulation of AFP gene expression before birth. All these results show that this serum-free culture of rat yolk sac explants is a valuable system for further investigating the action of natural compounds and pharmacological drugs on endodermal gene expression during the embryonic and fetal periods.

Animals↗

Endodermal sinus (yolk sac) tumor of oral cavity originating from gingiva.

Endodermal sinus tumor is a malignant germ cell tumor which usually arises in gonads. Extragonadal endodermal sinus tumors in the head and neck are very rare. Here a gingival endodermal sinus tumor is reported. The lesion demonstrated typical microscopic features of the endodermal sinus tumors of gonads. The tumor cells showed alpha-fetoprotein immunoreactivity in immunohistochemical evaluation. The serum alpha-fetoprotein level was high. This is the first gingival case in the related literature.

Biopsy↗

Specification of endoderm in the sea urchin embryo.

Expression of the endoderm specific gene Endo16, was used to monitor endoderm specification in developmentally arrested and in dissociated embryos. beta-APN treatment halts gastrulation, however, in two species of sea urchins Endo16 mRNA is still expressed, suggesting that specification of the endodermal lineage has taken place in these developmentally arrested embryos. Endo16 mRNA is not expressed in embryos dissociated at the 4-8-cell stage unless they are reassociated shortly after the 16-cell stage. Interestingly, dissociation after the 16-cell stage also results in a lack of Endo16 expression. Lithium is unable to rescue Endo16 expression in these dissociated embryos. These results indicate that early signaling events mediated by cell-cell contact are required for the initial specification and maintenance of the endoderm in the developing embryo.

Amino Acid Sequence↗

Drosophila endoderm development requires a novel homeobox gene which is a target of Wingless and Dpp signalling.

We have identified and cloned a novel type of homeobox gene that is composed of two homeodomains and is expressed in the Drosophila endoderm. Mutant analysis reveals that its activity is required at the foregut/midgut boundary for the development of the proventriculus. This organ regulates food passage from the foregut into the midgut and forms by the infolding of ectoderm and endoderm-derived tissues. The endodermal outer wall structure of the proventriculus is collapsed in the mutants leading to a failure of the ectodermal part to invaginate and build a functional multilayered organ. Lack-of-function and gain-of-function experiments show that the expression of this homeobox gene in the proventriculus endoderm is induced in response to Wingless activity emanating from the ectoderm/endoderm boundary whereas its expression in the central midgut is controlled by Dpp and Wingless signalling emanating from the overlying visceral mesoderm.

Alternative Splicing↗

Appearance of erythrocyte-like globules in the mouse visceral yolk sac endodermal cells on embryonic day 12, with special reference to blood islands.

The mouse visceral yolk sac (VYS) is widely known to play an important role as erythropoietic tissue during embryonic periods. Mouse VYS from embryonic days 9 to 12 was examined by light microscopy, electron microscopy and histochemical analysis with benzidine to detect the presence of hemoglobin with special reference to the development of VYS, the disappearance of the blood islands in VYS, and the appearance of a novel structure in the form of erythrocyte-like globules in VYS endodermal cells. The villous appearance of VYS became complicated by the development of VYS endodermal cells. The blood islands positive for the benzidine reaction were light microscopically detected on embryonic days 9, 10, and 11. They disappeared on embryonic day 12, however. Erythrocyte-like globules positive for the benzidine reaction were not observed in VYS endodermal cells on embryonic days 9, 10, and 11, but then appeared on embryonic day 12, by light and electron microscopy. Erythrocyte-like globules in VYS endodermal cells, which appear after the disappearance of blood islands in VYS, may participate in erythropoiesis during embryonic development.

Animals↗

Angiomotin regulates visceral endoderm movements during mouse embryogenesis.

In pregastrula stage mouse embryos, visceral endoderm (VE) migrates from a distal to anterior position to initiate anterior identity in the adjacent epiblast. This anterior visceral endoderm (AVE) is then displaced away from the epiblast by the definitive endoderm to become associated with the extra-embryonic ectoderm and subsequently contributes to the yolk sac. Little is known about the molecules that regulate this proximal displacement. Here we describe a role for mouse angiomotin (amot) in VE movements. amot expression is initially detected in the AVE and subsequently in the VE associated with the extra-embryonic ectoderm. Most amot mutant mice die soon after gastrulation with distinct furrows of VE located at the junction of the embryonic and extra-embryonic regions. Mutant analysis suggests that VE accumulation in these furrows is caused by defects in cell migration into proximal extra-embryonic regions, although distal-to-anterior movements associated with the epiblast, definitive endoderm formation, and anterior specification of the epiblast appear to be normal. These results suggest that amot acts within subregions of the VE to regulate morphogenetic movements that are required for embryo viability.

Amino Acid Sequence↗

Isolation and characterization of an endodermally derived, proteoglycan-like extracellular matrix molecule that may be involved in larval starfish digestive tract morphogenesis.

A monoclonal antibody, anti-Pisaster matrix-1 (anti-PM1) has been developed against an extracellular matrix antigen, Pisaster matrix-1 (PM1) found in embryos and larvae of the starfish Pisaster ochraceus. Pisaster matrix-1 was first observed in endodermal cells of the early gastrula, and shortly thereafter it was secreted into the blastocoel where it accumulated steadily during gastrulation. During the late gastrula stage it also appeared in the extracellular matrix (ECM) of the gut lumen. Immunogold electron microscopy with anti-PM1 revealed that PM1 was found in condensations of ECM associated with blastocoel matrix fibers, in the trans Golgi network, in Golgi-associated vesicles in endoderm and mesenchyme cells and throughout the ECM lining the digestive tract of late gastrula and bipinnaria larvae. When blastula or early gastrula stage embryos were grown in the presence of the PM1 antibody, archenteron elongation, bending and mouth formation failed to occur. Pisaster matrix-1 stained with alcian blue and its assembly could be disrupted with the common inhibitor of O-linked glycosaminoglycan assembly, beta-xyloside but not by tunicamycin. It was not sensitive to enzymes that degrade vertebrate proteoglycans. Pisaster matrix-1 is a large (600 kDa) proteoglycan-like glycosaminoglycan, secreted exclusively by endodermal and/or endodermally derived cells that may be necessary for morphogenesis of the mouth and digestive tract of Pisaster ochraceus embryos/larvae.

Animals↗

Fate mapping study of the endoderm in the posterior part of the 1.5-day-old chick embryo.

Various endodermal sites posterior to the caudal-most somite were marked in ovo with the vital dye Dil, and the fate of marked endoderm was analyzed after 2 or 3 days' reincubation. The endoderm in this area became gut epithelium posterior to the caudal jejunum and yolk sac. The area occupied by the cells that were to contribute to the dorsal part of the digestive tube lay centrally around the area overlaid by axial and paraxial mesoderm, with the preventral digestive area lying outside with considerable overlapping, which was surrounded by the preyolk sac area. During the formation of the posterior digestive tube, the endoderm was elongated anteroposteriorly to a considerable degree. Cells that contributed to the cloaca and those that produced descendants in the large intestine occupied similar areas posterior to the center of the sinus rhomboidalis, which were included in the pre-ileal area extending more anteriorly. Prejejunal cells generally localized in a more anterior position than pre-ileal cells. Pre-allantoic cells were located in a rather small area around the posterior primitive streak.

Animals↗

Identification of mature plasma cells in early rat yolk sac. A possible origin from the endodermal cell layer: immunohistochemistry and immunoelectron microscopic study.

Plasma cells play a pivotal role in the immune system and are responsible for the synthesis and release of immunoglobulins. Numerous in vitro culture experiments on the yolk sac demonstrated the generation of mature cells of the myeloid and lymphoid lineages under appropriate culture conditions. However, there are no reports describing the development of mature lymphoid cells in the yolk sac so far. For this reason, we undertook this study to investigate the development of antibody-containing plasma cells during early yolk sac haematopoiesis. Immunohistochemistry and immunoelectron microscopy were employed in the study. Results of this work demonstrated very weak immune staining for the intracytoplasmic IgA, IgG, and IgM at days 10 and 11 of embryonic life, while dark staining was obtained at 12 days. Positive staining was localized to the endodermal cell layer. Electron microscopic examinations revealed the existence of cells with the typical characteristics of plasma cells inside the endodermal cell layer, which may suggest their endodermal origin. To further verify the nature of these cells, intracytoplasmic immunoglobulins were demonstrated by immunoelectron microscopy. The present study demonstrated emergence of mature functioning plasma cells in early rat yolk sac. In a previous work we hypothesized the possibility of endodermal origin of yolk sac macrophages. This study adds additional evidence to support that hypothesis. The possible role of plasma cells in the yolk sac is discussed.

Animals↗

Parietal endoderm cell line from a rat blastocyst.

We have derived a cell line, RE1, from a pre-implantation rat blastocyst, resembling morphologically the L2 cell line from a parietal yolk sac carcinoma of the rat, as well as parietal endoderm cell lines of the mouse. The sub-cellular organization and epithelial characteristics of RE1 cells are described. The cells express cytokeratins of simple epithelia, and vimentin; and demonstrate synthesis of proteins of the extracellular matrix, such as laminin and collagen IV. Extensive Reichert's-like basement membrane is formed by RE1 cells when grown in suspension as aggregates. Cells have a microvillous surface morphology and abundant, rough endoplasmic reticulum which is swollen with apparent secretory material. These morphological and cytochemical features are characteristic of parietal endoderm cells in vivo, and the RE1 cell line is deduced to be rat parietal endoderm. In addition, RE1 cells were examined for expression of stage-specific embryonic antigens: cells reacted with antibody against SSEA-1/TEC-1 and EMA-1, constituting the first observation of parietal endoderm cells expressing the respective epitopes. RE1-cell monolayers did not generate transepithelial resistances or potential differences in vitro, consistent with their formation of leaky epithelia. Our observations on RE1-cell morphology and ultrastructure are consistent with the occurrence of epithelial-mesenchyme transitions in culture.

Animals↗