ABSTRACT
The insect midgut epithelium represents an interface between the internal and the external environment and it is the almost unique epithelial tissue by which these arthropods acquire nutrients. This epithelium is indeed able to produce digestive enzymes and to support vectorial transport of small organic nutrients, ions, and water. Moreover, it plays a key role in the defense against pathogenic microorganisms and in shaping gut microbiota. Another important midgut function is the ability to produce signaling molecules that regulate its own physiology and the activity of other organs. The two main mature cell types present in the midgut of all insects, i.e., columnar and endocrine cells, are responsible for these functions. In addition, stem cells, located at the base of the midgut epithelium, ensure the growth and renewal of the midgut during development and after injury. In insects belonging to specific orders, midgut physiology is deeply conditioned by the presence of unique cell types, i.e., goblet and copper cells, which confer peculiar features to this organ. This review reports current knowledge on the cells that form the insect midgut epithelium, focusing attention on their morphological and functional features. Notwithstanding the apparent structural simplicity of this organ, the properties of the cells make the midgut a key player in insect development and homeostasis.
Subject(s)
Digestive System/ultrastructure , Endoderm/ultrastructure , Insecta/anatomy & histology , AnimalsABSTRACT
The urethra within the human penile shaft develops via (1) an "Opening Zipper" that facilitates distal canalization of the solid urethral plate to form a wide urethral groove and (2) a "Closing Zipper" that facilitates fusion of the epithelial surfaces of the urethral folds. Herein, we extend our knowledge by describing formation of the human urethra within the glans penis as well as development of the prepuce. Forty-eight normal human fetal penile specimens were examined using scanning electron microscopy and optical projection tomography. Serial histologic sections were evaluated for morphology and immunohistochemical localization for epithelial differentiation markers: Cytokeratins 6, 7, 10, FoxA1, uroplakin and the androgen receptor. As the closing zipper completes fusion of the urethral folds within the penile shaft to form a tubular urethra (~ 13 weeks), canalization of the urethral plate continues in proximal to distal fashion into the glans penis to directly form the urethra within the glans without forming an open urethral groove. Initially, the urethral plate is attached ventrally to the epidermis via an epithelial seam, which is remodeled and eliminated, thus establishing mesenchymal confluence ventral to the glanular urethra. The morphogenetic remodeling involves the strategic expression of cytokeratin 7, FoxA1 and uroplakin in endodermal epithelial cells as the tubular glanular urethra forms. The most ventral epithelial cells of the urethral plate are pinched off from the glanular urethra and are reabsorbed into the epidermis ultimately losing expression of their markers, a process undoubtedly regulated by androgens. The prepuce initially forms on the dorsal aspect of the glans at approximately 12 weeks of gestation. After sequential proximal to distal remodeling of the ventral urethral plate along the ventral aspect of glans, the prepuce of epidermal origin fuses in the ventral midline.
Subject(s)
Cell Differentiation/genetics , Morphogenesis/genetics , Penis/ultrastructure , Urethra/ultrastructure , Endoderm/growth & development , Endoderm/metabolism , Endoderm/ultrastructure , Epithelial Cells/metabolism , Epithelial Cells/ultrastructure , Gene Expression Regulation, Developmental/genetics , Hepatocyte Nuclear Factor 3-alpha/genetics , Humans , Male , Penis/growth & development , Receptors, Androgen/genetics , Urethra/growth & development , Uroplakins/geneticsABSTRACT
BACKGROUND: Gastrulation is a complex orchestration of movements by cells that are specified early in development. Until now, classical convergent extension was considered to be the main contributor to sea urchin archenteron extension, and the relative contributions of cell divisions were unknown. Active migration of cells along the axis of extension was also not considered as a major factor in invagination. RESULTS: Cell transplantations plus live imaging were used to examine endoderm cell morphogenesis during gastrulation at high-resolution in the optically clear sea urchin embryo. The invagination sequence was imaged throughout gastrulation. One of the eight macromeres was replaced by a fluorescently labeled macromere at the 32 cell stage. At gastrulation those patches of fluorescent endoderm cell progeny initially about 4 cells wide, released a column of cells about 2 cells wide early in gastrulation and then often this column narrowed to one cell wide by the end of archenteron lengthening. The primary movement of the column of cells was in the direction of elongation of the archenteron with the narrowing (convergence) occurring as one of the two cells moved ahead of its neighbor. As the column narrowed, the labeled endoderm cells generally remained as a contiguous population of cells, rarely separated by intrusion of a lateral unlabeled cell. This longitudinal cell migration mechanism was assessed quantitatively and accounted for almost 90% of the elongation process. Much of the extension was the contribution of Veg2 endoderm with a minor contribution late in gastrulation by Veg1 endoderm cells. We also analyzed the contribution of cell divisions to elongation. Endoderm cells in Lytechinus variagatus were determined to go through approximately one cell doubling during gastrulation. That doubling occurs without a net increase in cell mass, but the question remained as to whether oriented divisions might contribute to archenteron elongation. We learned that indeed there was a biased orientation of cell divisions along the plane of archenteron elongation, but when the impact of that bias was analyzed quantitatively, it contributed a maximum 15% to the total elongation of the gut. CONCLUSIONS: The major driver of archenteron elongation in the sea urchin, Lytechinus variagatus, is directed movement of Veg2 endoderm cells as a narrowing column along the plane of elongation. The narrowing occurs as cells in the column converge as they migrate, so that the combination of migration and the angular convergence provide the major component of the lengthening. A minor contributor to elongation is oriented cell divisions that contribute to the lengthening but no more than about 15%.
Subject(s)
Gastrula/growth & development , Gastrulation/physiology , Morphogenesis/physiology , Sea Urchins/embryology , Animals , Cell Movement/genetics , Endoderm/growth & development , Endoderm/ultrastructure , Gastrula/ultrastructure , Sea Urchins/genetics , Sea Urchins/ultrastructureABSTRACT
How genetic programs generate cell-intrinsic forces to shape embryos is actively studied, but less so how tissue-scale physical forces impact morphogenesis. Here we address the role of the latter during axis extension, using Drosophila germband extension (GBE) as a model. We found previously that cells elongate in the anteroposterior (AP) axis in the extending germband, suggesting that an extrinsic tensile force contributed to body axis extension. Here we further characterized the AP cell elongation patterns during GBE, by tracking cells and quantifying their apical cell deformation over time. AP cell elongation forms a gradient culminating at the posterior of the embryo, consistent with an AP-oriented tensile force propagating from there. To identify the morphogenetic movements that could be the source of this extrinsic force, we mapped gastrulation movements temporally using light sheet microscopy to image whole Drosophila embryos. We found that both mesoderm and endoderm invaginations are synchronous with the onset of GBE. The AP cell elongation gradient remains when mesoderm invagination is blocked but is abolished in the absence of endoderm invagination. This suggested that endoderm invagination is the source of the tensile force. We next looked for evidence of this force in a simplified system without polarized cell intercalation, in acellular embryos. Using Particle Image Velocimetry, we identify posteriorwards Myosin II flows towards the presumptive posterior endoderm, which still undergoes apical constriction in acellular embryos as in wildtype. We probed this posterior region using laser ablation and showed that tension is increased in the AP orientation, compared to dorsoventral orientation or to either orientations more anteriorly in the embryo. We propose that apical constriction leading to endoderm invagination is the source of the extrinsic force contributing to germband extension. This highlights the importance of physical interactions between tissues during morphogenesis.
Subject(s)
Drosophila/embryology , Embryo, Nonmammalian/anatomy & histology , Endoderm/embryology , Gastrulation , Models, Anatomic , Morphogenesis , Animals , Biomarkers/metabolism , Cell Shape , Cell Size , Drosophila/genetics , Drosophila/metabolism , Drosophila/ultrastructure , Drosophila Proteins/genetics , Drosophila Proteins/metabolism , Embryo, Nonmammalian/metabolism , Embryo, Nonmammalian/ultrastructure , Endoderm/metabolism , Endoderm/ultrastructure , Green Fluorescent Proteins/genetics , Green Fluorescent Proteins/metabolism , Mechanical Phenomena , Membrane Fusion Proteins/genetics , Membrane Fusion Proteins/metabolism , Microscopy, Electron, Scanning/veterinary , Microscopy, Video/veterinary , Mutation , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/metabolism , Rheology , Time-Lapse Imaging/veterinary , Twist-Related Protein 1/genetics , Twist-Related Protein 1/metabolismABSTRACT
The freshwater shrimp Neocaridina heteropoda (Crustacea, Malacostraca, Decapoda) originates from Asia and is one of the species that is widely available all over the world because it is the most popular shrimp that is bred in aquaria. The structure and the ultrastructure of the midgut have been described using X-ray microtomography, transmission electron microscopy, light and fluorescence microscopes. The endodermal region of the alimentary system in N. heteropoda consists of an intestine and a hepatopancreas. No differences were observed in the structure and ultrastructure of males and females of the shrimp that were examined. The intestine is a tube-shaped organ and the hepatopancreas is composed of two large diverticles that are divided into the blind-end tubules. Hepatopancreatic tubules have three distinct zones - proximal, medial and distal. Among the epithelial cells of the intestine, two types of cells were distinguished - D and E-cells, while three types of cells were observed in the epithelium of the hepatopancreas - F, B and E-cells. Our studies showed that the regionalization in the activity of cells occurs along the length of the hepatopancreatic tubules. The role and ultrastructure of all types of epithelial cells are discussed, with the special emphasis on the function of the E-cells, which are the midgut regenerative cells. Additionally, we present the first report on the existence of an intercellular junction that is connected with the E-cells of Crustacea.
Subject(s)
Decapoda , Endoderm/cytology , Endoderm/ultrastructure , Gastrointestinal Tract/cytology , Gastrointestinal Tract/ultrastructure , Animals , Endoderm/embryology , Gastrointestinal Tract/embryologyABSTRACT
The medusa, Aurelia aurita (Scyphozoa, Cnidaria), is considered to be a cosmopolitan species with a worldwide distribution in most seas from the poles to the tropics. Cnidarian is thought to possess two tissue layers: endoderm (gastroderm) and ectoderm, which are separated by huge mesoglea in medusa. The basic morphology of medusa is similar in different populations. Previously we have determined a new protein "mesoglein" as one of the main components of mesoglea. Deduced amino acid sequence of mesoglein contains Zona Pellucida (ZP) domain. In this paper, we have comparied of mesoglein and its gene in medusa from three habitats (White Sea (WsA), Black Sea (BsA), Japonic Sea (JsA)). The set of the mesoglea protein bands after SDS-PAGE is similar in all samples. Nevertheless, JsA mesogleins' M(r) is 53-55 kDa, while WsA and BsA mesogleins have M(r) of 47 kDa. Antibodies raised against WsA mesoglein recognize only mesogleins with M(r) of 47 kDa, but not 53-55 kDa, both on immunoblot and immunocytochemistry. Mesogleal cells and elastic fibrils are stained intensively in the mesoglea both from WsA and BsA but not from JsA. The possibility of gene divergency was checked by PCR with primers specific for WsA mesoglein gene. PCR products of expected length obtained on polyA-cDNA template from mesogleal cells of WsA and BsA medusa but not on cDNA of JsA medusa. Our results evidence that there are two different species in genus Aurelia: Aurelia aurita inhabits White and Black Seas while Aurelia sp. inhabits Japonic Sea. This is consistent with findings of other recept molecular biological studies.
Subject(s)
Genetic Speciation , Proteins/genetics , Scyphozoa/classification , Animals , Antibodies/chemistry , Antibodies/isolation & purification , Blotting, Western , Ectoderm/ultrastructure , Endoderm/ultrastructure , Gene Expression , Genetics, Population , Guinea Pigs , Immunohistochemistry , Oceans and Seas , Polymerase Chain Reaction , Proteins/chemistry , Scyphozoa/genetics , Scyphozoa/ultrastructureABSTRACT
The poultry industry is a sector of agribusiness which represents an important role in the country's agricultural exports. Therefore, the study about embryogenesis of the domestic chicken (Gallus gallus domesticus) has a great economic importance. The aim of this study was to evaluate embryonic development of the endoderm in chicken (Gallus gallus domesticus). Forty fertilized eggs of domestic chickens, starting from the 1st day of gestation and so on until the 19 days of the incubation were collected from the Granja São José (Amparo, SP, Brazil). Embryos and fetus were fixed in 10% formaldehyde solution, identified, weighed, measured, and subjected to light and scanning electron microscopy. The endoderm originates the internal lining epithelium of the digestive, immune, respiratory systems, and the organs can be visualized from the second day (48 h) when the liver is formed. The formation of the digestive system was complete in the 12th day. Respiratory system organs begin at the fourth day as a disorganized tissue and undifferentiated. Their complete differentiation was observed at the 10 days of incubation, however, until the 19 days the syrinx was not observed. The formation of immune system at 10th day was observed with observation of the spleen, thymus, and cloacal bursa. The study of the organogenesis of the chicken based on germ layers is very complex and underexplored, and the study of chicken embryology is very important due the economic importance and growth of the use of this animal model studies such as genetic studies.
Subject(s)
Chick Embryo/embryology , Embryonic Development , Endoderm/embryology , Animals , Chick Embryo/anatomy & histology , Chick Embryo/ultrastructure , Chickens/anatomy & histology , Chickens/growth & development , Endoderm/anatomy & histology , Endoderm/ultrastructure , Liver/anatomy & histology , Liver/embryology , Liver/ultrastructure , Microscopy, Electron, Scanning , Spleen/anatomy & histology , Spleen/embryology , Spleen/ultrastructureABSTRACT
The metabolism of membrane phosphoinositides is critical for a variety of cellular processes. Phosphatidylinositol-3,5-bisphosphate [PtdIns(3,5)P(2)] controls multiple steps of the intracellular membrane trafficking system in both yeast and mammalian cells. However, other than in neuronal tissues, little is known about the physiological functions of PtdIns(3,5)P(2) in mammals. Here, we provide genetic evidence that type III phosphatidylinositol phosphate kinase (PIPKIII), which produces PtdIns(3,5)P(2), is essential for the functions of polarized epithelial cells. PIPKIII-null mouse embryos die by embryonic day 8.5 because of a failure of the visceral endoderm to supply the epiblast with maternal nutrients. Similarly, although intestine-specific PIPKIII-deficient mice are born, they fail to thrive and eventually die of malnutrition. At the mechanistic level, we show that PIPKIII regulates the trafficking of proteins to a cell's apical membrane domain. Importantly, mice with intestine-specific deletion of PIPKIII exhibit diarrhea and bloody stool, and their gut epithelial layers show inflammation and fibrosis, making our mutants an improved model for inflammatory bowel diseases. In summary, our data demonstrate that PIPKIII is required for the structural and functional integrity of two different types of polarized epithelial cells and suggest that PtdIns(3,5)P(2) metabolism is an unexpected and critical link between membrane trafficking in intestinal epithelial cells and the pathogenesis of inflammatory bowel disease.
Subject(s)
Endoderm/metabolism , Intestinal Mucosa/metabolism , Phosphatidylinositol 3-Kinases/genetics , Viscera/metabolism , Animals , Cells, Cultured , Embryo, Mammalian/embryology , Embryo, Mammalian/metabolism , Embryo, Mammalian/ultrastructure , Embryonic Stem Cells/metabolism , Endoderm/embryology , Endoderm/ultrastructure , Female , Gene Expression Profiling , Inflammatory Bowel Diseases/genetics , Inflammatory Bowel Diseases/metabolism , Intestines/ultrastructure , Male , Mice , Mice, Inbred C57BL , Mice, Knockout , Microscopy, Confocal , Microscopy, Electron , Oligonucleotide Array Sequence Analysis , Phosphatidylinositol 3-Kinases/metabolism , Phosphatidylinositol Phosphates/metabolism , RNA Interference , Reverse Transcriptase Polymerase Chain Reaction , Time Factors , Viscera/embryology , Viscera/ultrastructureABSTRACT
GATA-6 is a zinc-finger transcription factor essential for early embryogenesis. Ablation of GATA-6 in mice impairs endoderm differentiation and causes apoptosis of epiblast cells. The endoderm defects have been attributed to the loss of HNF4, disabled-2, and GATA-4. However, the mechanisms underlying epiblast apoptosis are unclear. In this study we used mouse embryonic stem cell-derived embryoid bodies (EBs) as a model for peri-implantation development and found that ablation of GATA-6 causes massive apoptosis during EB differentiation. Endoderm grafting experiments and ectopic basement membrane (BM) assembly suggest that both BM and non-BM factors contribute to cell survival. Furthermore, the increased cell death in mutant EBs is accompanied by reduced expression of bone morphogenetic protein 2 (BMP-2). Chromatin immunoprecipitation reveals direct binding of GATA-6 to the Bmp2 promoter. Treatment of the mutant EBs with BMP-2 markedly suppresses apoptosis, whereas stable overexpression of the BMP antagonist noggin or a dominant-negative BMP receptor in normal EBs leads to increased apoptosis. Last, activation of SMAD1/5 by phosphorylation is significantly inhibited in the absence of GATA-6, and this is reversed by exogenous BMP-2. Treatment of normal EBs with SMAD phosphorylation inhibitor increases apoptosis. Collectively these results suggest that GATA-6 promotes cell survival by regulating endoderm expression of BMP-2 and BM during embryonic epithelial morphogenesis.
Subject(s)
Bone Morphogenetic Protein 2/metabolism , Cell Differentiation , Embryonic Stem Cells/metabolism , GATA6 Transcription Factor/metabolism , Animals , Apoptosis/drug effects , Apoptosis/genetics , Bone Morphogenetic Protein 2/genetics , Bone Morphogenetic Protein 2/pharmacology , Cell Line , Cell Survival , Embryoid Bodies/cytology , Embryoid Bodies/metabolism , Embryonic Stem Cells/cytology , Endoderm/metabolism , Endoderm/ultrastructure , GATA6 Transcription Factor/genetics , Gene Expression Profiling , Germ Layers/metabolism , Immunoblotting , Mice , Microscopy, Electron , Microscopy, Fluorescence , Mutation , Phosphorylation , Promoter Regions, Genetic/genetics , Protein Binding , Reverse Transcriptase Polymerase Chain Reaction , Smad Proteins/genetics , Smad Proteins/metabolism , Time Factors , Up-RegulationABSTRACT
Two lineages of endoderm develop during mammalian embryogenesis, the primitive endoderm in the pre-implantation blastocyst and the definitive endoderm at gastrulation. This complexity of endoderm cell populations is mirrored during pluripotent cell differentiation in vitro and has hindered the identification and purification of the definitive endoderm for use as a substrate for further differentiation. The aggregation and differentiation of early primitive ectoderm-like (EPL) cells, resulting in the formation of EPL-cell derived embryoid bodies (EPLEBs), is a model of gastrulation that progresses through the sequential formation of primitive streak-like intermediates to nascent mesoderm and more differentiated mesoderm populations. EPL cell-derived EBs have been further analysed for the formation of definitive endoderm by detailed morphological studies, gene expression and a protein uptake assay. In comparison to embryoid bodies derived from ES cells, which form primitive and definitive endoderm, the endoderm compartment of embryoid bodies formed from EPL cells was comprised almost exclusively of definitive endoderm. Definitive endoderm was defined as a population of squamous cells that expressed Sox17, CXCR4 and Trh, which formed without the prior formation of primitive endoderm and was unable to endocytose horseradish peroxidase from the medium. Definitive endoderm formed in EPLEBs provides a substrate for further differentiation into specific endoderm lineages; these lineages can be used as research tools for understanding the mechanisms controlling lineage establishment and the nature of the transient intermediates formed. The similarity between mouse EPL cells and human ES cells suggests EPLEBs can be used as a model system for the development of technologies to enrich for the formation of human ES cell-derived definitive endoderm in the future.
Subject(s)
Cell Culture Techniques/methods , Cell Differentiation/physiology , Embryoid Bodies/ultrastructure , Endoderm/ultrastructure , Mesoderm/ultrastructure , Pluripotent Stem Cells/ultrastructure , Primitive Streak/ultrastructure , Animals , DNA Primers/genetics , Flow Cytometry , Gene Expression Profiling , Horseradish Peroxidase/pharmacokinetics , Mice , Microscopy, Electron, Scanning , Microscopy, Electron, Transmission , Pluripotent Stem Cells/metabolism , Real-Time Polymerase Chain Reaction , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
The Crumbs family of transmembrane proteins has an important role in the differentiation of the apical membrane domain in various cell types, regulating such processes as epithelial cell polarization. The mammalian Crumbs protein family is composed of three members. Here, we inactivated the mouse Crb2 gene with gene-targeting techniques and found that the protein is crucial for early embryonic development with severe abnormalities appearing in Crb2-deficient embryos at late-gastrulation. Our findings indicate that the primary defect in the mutant embryos is disturbed polarity of the epiblast cells at the primitive streak, which affects epithelial to mesenchymal transition (EMT) during gastrulation, resulting in impaired mesoderm and endoderm formation, and embryonic lethality by embryonic day 12.5. These findings therefore indicate a novel role for the Crumbs family of proteins.
Subject(s)
Cell Polarity/physiology , Endoderm/embryology , Epithelial-Mesenchymal Transition/physiology , Gastrulation/physiology , Membrane Proteins/biosynthesis , Mesoderm/embryology , Animals , Embryo Loss/genetics , Embryo Loss/metabolism , Embryo Loss/pathology , Endoderm/ultrastructure , Membrane Proteins/genetics , Mesoderm/ultrastructure , Mice , Mice, Mutant StrainsABSTRACT
The Drosophila larval and adult midguts are derived from two populations of endodermal progenitors that separate from each other in the early embryo. As larval midgut cells differentiate into an epithelial layer, adult midgut progenitors (AMPs) remain as small clusters of proliferating, undifferentiated cells attached to the basal surface of the larval gut epithelium. During the first few hours of metamorphosis, AMPs merge into a continuous epithelial tube that overgrows the larval layer and differentiates into the adult midgut; at the same time, the larval midgut degenerates. As shown in this paper, there is a second, transient pupal midgut that develops from the AMPs at the beginning of metamorphosis and that intercalates between the adult and larval midgut epithelia. Cells of the transient pupal midgut form a multilayered tube that exhibits signs of differentiation, in the form of septate junctions and rudimentary apical microvilli. Some cells of the pupal midgut develop as endocrine cells. The pupal midgut remains closely attached to the degenerating larval midgut cells. Along with these cells, pupal midgut cells are sequestered into the lumen where they form the compact "yellow body." The formation of a pupal midgut has been reported from several other species and may represent a general feature of intestinal metamorphosis in insects.
Subject(s)
Drosophila melanogaster/growth & development , Intestines/growth & development , Metamorphosis, Biological , Animals , Drosophila melanogaster/ultrastructure , Endoderm/growth & development , Endoderm/ultrastructure , Epithelium/growth & development , Epithelium/ultrastructure , Intestines/ultrastructure , Larva/ultrastructure , Pupa/growth & development , Pupa/ultrastructureABSTRACT
The present study gives a detailed ultrastructural description of equine conceptuses at Day 14 (n = 2) and Day 16 (n = 3) after ovulation. Whereas on Day 14 only primitive structures were seen, on Day 16 neurulation and formation of mesodermal somites had taken place. The ectoderm of the embryo itself and the surrounding trophoblast ectodermal cells were characterised by specific cell surface differentiations. At the embryonic ectodermal cell surface (14 and 16 days) remarkable protruded and fused cytoplasmic projections were seen, typically associated with macropinocytotic events involved in macromolecule and fluid uptake. This finding adds an important point to the expansion mode of the hypotone equine conceptus, which is characterised by 'uphill' fluid uptake. Numerous microvilli and coated endocytotic pits at the apical trophoblast membrane emphasised its absorptive character. Endodermal cells were arranged loosely with only apically located cellular junctions leaving large intercellular compartments. At the border of the embryonic disc apoptotic cells were regularly observed indicating high remodelling activities in this area. Conspicuous blister-like structures between ectoderm and mesoderm were seen in the trilaminar part of Day-14 and -16 conceptuses. These were strictly circumscribed despite not being sealed by cellular junctions between germinal layers. It is possible that these blisters are involved in embryo positioning; however, further studies are needed to verify this.
Subject(s)
Embryo, Mammalian/ultrastructure , Horses/embryology , Animals , Ectoderm/ultrastructure , Embryonic Development , Endoderm/ultrastructure , Female , Gestational Age , Mesoderm/ultrastructure , Microscopy, Electron, Transmission , Pregnancy , Somites/ultrastructureABSTRACT
The expression pattern of caveolin-1 and the distribution of caveolae in the murine placental labyrinth and visceral yolk sac have been determined. Immunoblot analysis demonstrates that both placenta and yolk sac express the protein caveolin-1. Immunofluorescence microscopy was used to determine which cell types in the placental labyrinth and yolk sac express caveolin-1. In yolk sac, detectable caveolin-1 was restricted to endothelial cells and smooth muscle cells of the vitelline vasculature and to mesothelial cells. Endoderm, the major cell type in the yolk sac, does not express caveolin-1 as assessed by this assay. In the labyrinth region of the placenta, endothelial cells express caveolin-1 but this protein was not detectable in any of the three trophoblast layers. These tissues were also examined by electron microscopy to determine which cell types contain the specialized plasma membrane microdomains known as caveolae. Morphologically detectable caveolae were present in endothelial and smooth muscle cells, as well as mesothelial cells of the yolk sac and in endothelial cells of the placental labyrinth. Neither endodermal cells of the yolk sac nor trophoblastic cells in the placental labyrinth contained caveolae-like structures. We conclude that caveolin-1 and caveolae have restricted distribution in the murine placenta and yolk sac and that this parallels the situation in human placenta.
Subject(s)
Caveolae/ultrastructure , Caveolin 1/metabolism , Placenta/metabolism , Placenta/ultrastructure , Pregnancy Proteins/metabolism , Yolk Sac/metabolism , Yolk Sac/ultrastructure , Animals , Cell Polarity , Endoderm/metabolism , Endoderm/ultrastructure , Endothelial Cells/metabolism , Endothelial Cells/ultrastructure , Female , Fluorescent Antibody Technique, Indirect , Humans , Mice , Mice, Inbred BALB C , Microscopy, Confocal , Microscopy, Fluorescence , Myocytes, Smooth Muscle/metabolism , Myocytes, Smooth Muscle/ultrastructure , Pregnancy , Protein Transport , Species Specificity , Trophoblasts/metabolism , Trophoblasts/ultrastructureABSTRACT
Ectodermal contribution to the induction of pharyngeal teeth that form in the endodermal territory of the oropharyngeal cavity in some teleost fishes has been a matter of considerable debate. To determine the role of ectodermal cell signaling in scale and tooth formation and thereby to gain insights in evolutionary origin of teeth, we analyzed scales and teeth in rs-3 medaka mutants characterized by reduced scale numbers due to aberrant splicing of the ectodysplasin-A receptor (edar). Current data show that, in addition to a loss of scales (83% reduction), a drastic loss of teeth occurred in both oral (43.5% reduction) and pharyngeal (73.5% reduction) dentitions in rs-3. The remaining scales of rs-3 were irregular in shape and nearly 3 times larger in size relative to those of the wild-type. In contrast, there was no abnormality in size and shape in the remaining teeth of rs-3. In wild-type medaka embryos, there was a direct contact between the surface ectoderm and rostral endoderm in pharyngeal regions before the onset of pharyngeal tooth formation. However, there was no sign of ectodermal cell migration in the pharyngeal endoderm and hence no direct evidence of any ectodermal contribution to pharyngeal odontogenesis. These data suggest differential roles for Eda-Edar signaling in the induction and growth of scales and teeth and support the intrinsic odontogenic competence of the rostral endoderm in medaka.
Subject(s)
Animal Structures/anatomy & histology , Biological Evolution , Oryzias/anatomy & histology , Oryzias/genetics , Pharynx/anatomy & histology , Receptors, Ectodysplasin/genetics , Tooth/anatomy & histology , Animals , Ectoderm/anatomy & histology , Ectoderm/ultrastructure , Embryo, Nonmammalian/ultrastructure , Endoderm/anatomy & histology , Endoderm/ultrastructure , Female , Male , Mutation/genetics , Oryzias/embryology , Pharynx/diagnostic imaging , Phenotype , Tomography, X-Ray Computed , Tooth/diagnostic imagingABSTRACT
Accumulating evidence demonstrates that cilia play important roles in a variety of processes in embryogenesis. For functional survey of larval cilia at the cellular level, we exploited the simple cell organization of tadpole larvae in the ascidian Ciona intestinalis. Immunofluorescent microscopy showed distribution of cilia not only in previously described tissues but also in a subpopulation of ependymal cells in the sensory vesicle, gut primordium, papillae, apical trunk epidermal neurons, and the endodermal strand. Transmission electron microscopy revealed a variety of axonemal structures, including a 9+0 structure similar to vertebrate primary cilia, a 9+0 structure with electron-dense materials in the center, a 9+2 structure with no dynein arms, and an axoneme with a disorganized structure at the distal end. Extensive description of cilia in the present study gives important insights into the evolution of the ciliary structure and provides a basis for analysis of ciliary functions in establishment of chordate body plan.
Subject(s)
Cilia/ultrastructure , Ciona intestinalis/embryology , Animals , Biological Evolution , Cilia/physiology , Endoderm/ultrastructure , Ependyma/ultrastructure , Larva/ultrastructure , Microscopy, Electron, Transmission , Microscopy, Fluorescence , Neural Tube/ultrastructure , Photoreceptor Cells/ultrastructure , Sensory Receptor Cells/ultrastructureABSTRACT
We demonstrate that conditional ablation of the homeobox transcription factor Cdx2 from early endoderm results in the replacement of the posterior intestinal epithelium with keratinocytes, a dramatic cell fate conversion caused by ectopic activation of the foregut/esophageal differentiation program. This anterior homeotic transformation of the intestine was first apparent in the early embryonic Cdx2-deficient gut by a caudal extension of the expression domains of several key foregut endoderm regulators. While the intestinal transcriptome was severely affected, Cdx2 deficiency only transiently modified selected posterior Hox genes and the primary enteric Hox code was maintained. Further, we demonstrate that Cdx2-directed intestinal cell fate adoption plays an important role in the establishment of normal epithelial-mesenchymal interactions, as multiple signaling pathways involved in this process were severely affected. We conclude that Cdx2 controls important aspects of intestinal identity and development, and that this function is largely independent of the enteric Hox code.
Subject(s)
Epithelium/metabolism , Homeodomain Proteins/metabolism , Intestines/embryology , Mesoderm/metabolism , Signal Transduction , Trans-Activators/metabolism , Animals , Body Patterning , CDX2 Transcription Factor , Cell Differentiation , Cell Proliferation , Endoderm/cytology , Endoderm/embryology , Endoderm/metabolism , Endoderm/ultrastructure , Epithelium/ultrastructure , Esophagus/cytology , Esophagus/metabolism , Gene Deletion , Gene Expression Regulation, Developmental , Homeodomain Proteins/genetics , Intestinal Mucosa/metabolism , Intestines/cytology , Intestines/ultrastructure , Keratinocytes/cytology , Keratinocytes/metabolism , Mesoderm/cytology , Mesoderm/ultrastructure , Mice , Mice, Mutant Strains , Microvilli/metabolism , Microvilli/ultrastructure , Organ Specificity , Transcription Factors/metabolismABSTRACT
To investigate formation of the three primary germ layers in mouse embryoid bodies (EBs), we observed changes in structure and gene expression over a 7-day culture period. We compared these changes using two methods for EB formation: hanging drop (HD) and static suspension culture (SSC). Light microscopy showed that a stratified columnar epithelial layer developed on the surface of EBs formed using the HD method. From Day 3 in culture, ultrastructural changes occurred in the aligned cellular membranes. Condensation of actin filaments was followed by formation of complicated adherent junctions and dilatation of intercellular canaliculi containing well-developed microvilli. These changes were more marked in EBs formed by the HD method than the SSC method. On Day 5 of culture, Brachyury gene expression, a marker for mesoderm formation, was detected only with the HD method. Nestin, an ectoderm marker, and Foxa2, an endoderm marker, were expressed with both methods. These results suggest that in EBs formed with the HD method, actin formation and Brachyury gene expression mark the transition from two to three primary germ layers. Additionally, the HD method promotes more rapid and complete development of mouse EBs than does the SSC method. While the SSC method is simple and easy to use, it needs improvement to form more complete EBs.
Subject(s)
Embryonic Development/genetics , Embryonic Stem Cells/physiology , Embryonic Stem Cells/ultrastructure , Gene Expression Regulation, Developmental , Actin Cytoskeleton/physiology , Actin Cytoskeleton/ultrastructure , Animals , Biomarkers , Cell Line , Ectoderm/embryology , Ectoderm/physiology , Ectoderm/ultrastructure , Endoderm/embryology , Endoderm/physiology , Endoderm/ultrastructure , Intercellular Junctions/physiology , Intercellular Junctions/ultrastructure , Mesoderm/embryology , Mesoderm/physiology , Mesoderm/ultrastructure , Mice , Microscopy, Electron, TransmissionABSTRACT
Coral bleaching is a major contributor to the global declines of coral reefs. This phenomenon is characterized by the loss of symbiotic algae, their pigments or both. Despite wide scientific interest, the mechanisms by which bleaching occurs are still poorly understood. Here we report that the removal of the symbiont during light and temperature stress is achieved using the host's cellular autophagic-associated machinery. Host cellular and subcellular morphologies showed increased vacuolization and appearance of autophagic membranes surrounding a variety of organelles and surrounding the symbiotic algae. Markers of autophagy (Rab 7 and LAS) corroborate these observations. Results showed that during stress the symbiont vacuolar membrane is transformed from a conduit of nutrient exchange to a digestive organelle resulting in the consumption of the symbiont, a process we term symbiophagy. We posit that during a stress event, the mechanism maintaining symbiosis is destabilized and symbiophagy is activated, ultimately resulting in the phenomenon of bleaching. Symbiophagy may have evolved from a more general primordial innate intracellular protective pathway termed xenophagy.
Subject(s)
Anthozoa/cytology , Anthozoa/metabolism , Autophagy , Symbiosis , Animals , Anthozoa/radiation effects , Anthozoa/ultrastructure , Autophagy/radiation effects , Biomarkers/metabolism , Endoderm/radiation effects , Endoderm/ultrastructure , Eukaryota/radiation effects , Eukaryota/ultrastructure , Light , Symbiosis/radiation effects , TemperatureABSTRACT
N eoturris breviconis (Anthomedusae) has a nerve plexus in the walls of its endodermal canals. The plexus is distinct from the ectodermal nerve plexuses supplying the radial and circular muscles in the ectoderm and no connections have been observed between them. Stimulation of the endodermal plexus evokes electrical events recorded extracellularly as "E" potentials. These propagate through all areas where the plexus has been shown by immunohistology to exist and nowhere else. When Neoturris is ingesting food, trains of "E" potentials propagate down the radial canals to the margin and cause inhibition of swimming. This response is distinct from the inhibition of swimming associated with contractions of the radial muscles but both may play a part in feeding and involve chemoreceptors. Preliminary observations suggest that the "E" system occurs in other medusae including Aglantha digitale (Trachymedusae) where the conduction pathway was previously thought to be an excitable epithelium.