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In vitro modeling of paraxial and lateral mesoderm differentiation reveals early reversibility.

Endothelial cells (ECs) are thought to be derived mainly from the vascular endothelial growth factor receptor 2 (VEGFR-2)+ lateral mesoderm during early embryogenesis. In this study, we specified several pathways for EC differentiation using a murine embryonic stem (ES) cell differentiation culture system that is a model for cellular processes during early embryogenesis. Based on the results of in vitro fate analysis, we show that, in the main pathway, committed ECs are differentiated through the VEGFR-2+ platelet-derived growth factor receptor alpha (PDGFR-alpha)- single-positive (VSP) population that is derived from the VEGFR-2+ PDGFR-alpha+ double-positive (DP) population. This major differentiation course was also confirmed using DNA microarray analysis. In addition to this main pathway, however, ECs also can be generated from the VEGFR-2- PDGFR-alpha+ single-positive (PSP) population, which represents the paraxial mesodermal lineage and is also derived from the DP population. Our results strongly suggest that, even after differentiation from the common progenitor DP population into the VSP and PSP populations, these two populations continue spontaneous switching of their surface phenotype, which results in switching of their eventual fates. The rate of this interlineage conversion between VSP and PSP is unexpectedly high. Because of this potential to undergo fate switch, we conclude that ECs can be generated via multiple pathways in in vitro ES cell differentiation.

Animals↗

Extrauterine mesodermal (müllerian) adenosarcoma. A case report.

Extrauterine mesodermal (müllerian) adenosarcomas have only recently been described, and this is the first reported case from Australia. These tumours fall within the category of common epithelial tumours' in the World Health Organisation (W.H.O.) classification of ovarian tumours and comprise benign looking epithelial structures (glands, papillae) in association with sarcomatous stroma. They are thus distinct from malignant mesodermal mixed tumours in which both epithelial and stromal elements are cytologically malignant.

Adult↗

Mesoderm induction in Xenopus laevis distinguishes between the various TGF-beta isoforms.

Induction of mesoderm in ectodermal explants of Xenopus laevis blastula embryos had previously been shown to respond selectively to TGF-beta 2, with TGF-beta s 1 and 5 having no activity in this assay. As TGF-beta s 1, 2, and 3 are frequently coexpressed in tissues, we wished to examine the activity of TGF-beta 3 relative to that of TGF-beta s 1 and 2 in this assay as well as in other in vitro assays. We report here that when the activity of recombinant TGF-beta 3 is normalized to that of TGF-beta 1 in the assay for growth inhibition in CCL-64 cells, it is also equal to that of TGF-beta 1 in assays for stimulation of both anchorage-independent growth of rat NRK cells and chemotaxis of human monocytes. In contrast, in the assay for mesoderm induction, recombinant TGF-beta 3 is 10-fold more active than TGF-beta 2, inducing expression of muscle specific alpha-actin at concentrations as low as 1 ng/ml. These results suggest that more complex systems, in contrast to individual cell types, may respond selectively to the various TGF-beta isoforms and that there might be biological consequences of TGF-beta isoform switching in vivo.

Animals↗

Brachyury is expressed by human teratocarcinoma cells in the absence of mesodermal differentiation.

Reverse transcription-PCR and Northern and Western blot analyses indicate that mRNA and protein encoded by the Brachyury gene are expressed by the pluripotent human embryonal carcinoma cell line NTERA2 and are only modestly down-regulated during retinoic acid-induced differentiation. This differentiation occurs along a neural lineage, with no obvious evidence of the formation of mesodermal derivatives. Several other human embryonal carcinoma cell lines that do not differentiate, a yolk sac carcinoma cell line and two choriocarcinoma cell lines, also express readily detectable levels of Brachyury mRNA and protein. Thus, in human teratocarcinomas, Brachyury expression is not necessarily an indicator of commitment to mesodermal differentiation.

Cell Differentiation↗

Organogenesis and tissue regeneration of fallopian tube: a desired metaplastic transformation of mesodermal stem cells in live animal models (dogs).

The capacity of stem cells of peritonium of mesodermal origin to undergo metaplastic transformation and form different tissues developed from mesoderm germ layer is exploited with ulterior motive to use it in the management of human diseases. The excised fallopian tube was replaced with a tube on a stent constructed from autogenous peritoneum from a suitable donor site. The effect of the surroundings environment of the new tissue system to which the peritoneum stem cells are now exposed was studied for 3, 6 and 12 months period in live animal models. The gross and histological studies revealed development of all the component of the wall of the fallopian tube. The lumen of the constructed peritoneal tube was well preserved in its whole length including the anastomotic sites. The scientific rationale of the working hypothesis on which the work is based, is discussed.

Anastomosis, Surgical↗

Making mesoderm--upstream and downstream of Xbra.

The mesoderm of the amphibian embryo is formed through an inductive interaction in which cells of the vegetal hemisphere of the embryo act on overlying equatorial cells. My laboratory is studying the Brachyury gene, which plays a key role in this interaction, being both necessary and sufficient for normal mesoderm formation. In this article I describe our attempts to understand how Xenopus Brachyury (Xbra) is activated in the right cells at the right time, and then to understand how Xbra exerts its effects.

Animals↗

Incidental finding of malignant mixed mesodermal tumor at hysterectomy for uterine prolapse. A case report.

BACKGROUND: The finding of unanticipated pathology in a uterus after vaginal hysterectomy for prolapse is uncommon. CASE: An incidental small malignant mixed mesodermal tumor was found at vaginal hysterectomy in a 68-year-old woman. CONCLUSION: A MED-LINE search found no other reported cases of malignant mixed mesodermal tumor in a patient undergoing vaginal hysterectomy for uterine prolapse. Unexpected endometrial and cervical lesions will be discovered occasionally after hysterectomy for benign disease.

Aged↗

Mesoderm formation by isolated and cultivated 8-cell stage blastomeres of the teleost, Leucopsarion ptersii (shiro-uo).

Isolation of cleavage-stage blastomeres and the study of their developmental potential has been used extensively for analyzing the mechanisms of embryogenesis in vertebrates, including amphibians and echinoderms. We devised a method to isolate 8-cell stage blastomeres in the teleost, shiro-uo, by utilizing its unique cleavage pattern of the horizontal 3rd cleavage plane. Removal of all the upper blastomeres at the 8-cell stage allowed almost normal embryogenesis from the remaining lower blastomeres and yolk cell mass. Isolated upper or lower blastomeres formed vesicles and spherical bodies, which later showed morphological changes during cultivation. Mesoderm formation was detected not only in the cultivated lower blastomeres or whole blastomeres but also in the upper blastomeres isolated from the yolk cell mass at the 8-cell stage, although at a lower frequency than the lower blastomeres. These results indicated the presence of very early signaling for mesoderm induction, which is independent from the currently postulated signals from the yolk syncytial layer at later stages. This also indicated non-equivalence or differentiation of the blastomeres from the very early cleavage stage in teleost embryos.

Amino Acid Sequence↗

Cell interactions and mesodermal cell fates in the sea urchin embryo.

Cell interactions during gastrulation play a key role in the determination of mesodermal cell fates in the sea urchin embryo. An interaction between primary and secondary mesenchyme cells (PMCs and SMCs, respectively), the two principal populations of mesodermal cells, regulates the expression of SMC fates. PMCs are committed early in cleavage to express a skeletogenic phenotype. During gastrulation, they transmit a signal that suppresses the skeletogenic potential of a subpopulation of SMCs and directs these cells into an alternative developmental pathway. This review summarizes present information concerning the cellular basis of the PMC-SMC interaction, as analyzed by cell transplantation and ablation experiments, fluorescent cell labeling methods and the use of cell type-specific molecular markers. The nature and stability of SMC fate switching, the timing of the PMC-SMC interaction and its quantitative characteristics, and the lineage, numbers and normal fate of the population of skeletogenic SMCs are discussed. Evidence is presented indicating that PMCs and SMCs come into direct filopodial contact during the late gastrula stage, when the signal is transmitted. Finally, evolutionary questions raised by these studies are briefly addressed.

Animals↗

[A strategy for immortalizing lines committed to endoderm, neuroectoderm or mesoderm from mouse teratocarcinoma].

With the aim of immortalizing embryonic cells fixed at early embryonic stages, various plasmids carrying the SV40 early region were introduced into the mouse embryonal carcinomas (EC) F9 and 1003. Only the construction PK4, in which the SV40 oncogenes are placed under the control of the adenovirus E1A promoter, led to the immortalization of the cells at the onset of differentiation. Clones corresponding to committed precursors of each embryonic lineage (neuroectoderm, mesoderm and endoderm) were then selected with high efficiency according to the following strategy: selection of immature cells which: have lost EC cell markers, keep a stable phenotype, are immortalized by the expression of the SV40 oncogenes and are still able to differentiate along a restricted lineage in vitro or in vivo. Examples of an endodermal precursor (H7) which differentiates into extraembryonic and embryonic endoderm, of a neuroectodermic clone (ICII) committed to a serotoninergic differentiation, and of a mesodermal osteogenic clone (CI) which gives rise to bone in vivo and in vitro, are given.

Adenoviridae↗

Effects of cytochalasin B on cell to cell adhesion and cellular shape of embryo mesoderm cells in vitro.

The effects of cytochalasin B on chick embryo mesoderm cells cultured in vitro were studied by scanning electron microscopy. The untreated cells showed numerous filopodia and lamellipodia and they were flattened onto the coverslip. Several cellular clusters were observed in each culture. The treated cells did not show filopodia, they had a rounded shape and cellular clusters were not present on the coverslip. These alterations are discussed on the basis of the actions of cytochalasin B on the micrcfilamentous cytoskeleton and the role proposed for microfilaments in filopodia formation, cellular shape and locomotion. It was observed that the mechanisms of cell to cell adhesion of chick embryo mesoderm cells were not the same as the mechanisms of cell adhesion to artificial substrates.

Animals↗

Dynamics of the control of body pattern in the development of Xenopus laevis. III. Timing and pattern after u.v. irradiation of the egg and after excision of presumptive head endo-mesoderm.

A series of Xenopus egg batches has been exposed to doses of u.v. (2537A) light on the vegetal hemisphere at precleavage stages, calculated to result in a range of minimal axial deficiency syndromes in the developing larvae. At the time of onset of gastrulation in synchronously fertilized but non-irradiated batch members, each experimental group was regularly scanned so that small subsamples of embryos could be set aside as showing particular, progressive degrees of delay in onset of the visible gastrulation movements. Such sampling was found to have preselected embryos showing generally progressive degrees of pattern impairment at larval stages, and this observation was extended by histological examination of the anterior axial anatomy. Such examination was also made of the least abnormal-looking members of a series of larvae resulting from excision of the presumptive head endo-mesoderm, traditionally called 'the organizer', from stage-10 gastrulae (Cooke, 1975). The results support the notion that production of the most anterior endo-mesodermal pattern parts (and of their inductive capacities in giving rise to the brain pattern) occurs only in material whose timing, in the onset of gastrulation activity, is close to the normal onset time after fertilization. Either an early failure of the egg to generate a location with the 'position value' corresponding with this extreme of the pattern, or the much later excision of the region from a physiologically normal gastrula, results in a system of pattern formation permanently truncated at its apical (head and dorsal) end. There is no evidence for any dynamic, in the system ascribing position value, that will cause regulative restoration of this cellular state (the most extreme 'activation' for development) in response to its absence after precleavage stages. An earlier statement (Cooke, 1975) that this could occur was based upon inadequate analysis of larvae with an often misleading external anatomy. The present results are discussed as supporting the overall view of the early Xenopus patterning system that has been developed in the previous two papers of the series.

Animals↗

[Differentiation behavior of the mesodermal germinal sone of differently old pregastrulation stages of Ambystoma mexicanum. Cultivation of untreated and lithium-treated isolates].

Isolates from the dorsal, lateral and ventral area of the marginal zone are showing differentiation capacities already in the stage of early morula. The composition of the differentiation pattern alters: ecto- and mesodermal tissues are realized until the late morula. Beyond these, endodermal structures are visible with the stage of early blastula. After a mild Li treatment of the isolates the differentiation pattern of the marginal zone shows significant modifications. Ectodermal organs are registered to a lesser degree. Gut-structures are already visible within the stage of middle morula. The capacity for gut-differentiation is not restricted to only the lateral and ventral areas but is also observed in the dorsal region of the marginal zone. The results of experiments are discussed on the basis of Nieuwkoop's formation of the mesoderm in urodelean amphibians.

Ambystoma↗

Identification and distribution of gap junctions in the mesoderm of the developing chick limb bud.

Sub-ridge, core, anterior and posterior borders of mesoderm were dissected from stages 22-24 chick wing buds to investigate whether structures for intercellular coupling develop between mesenchymal cells. Fine structure was examined using techniques of transmission electron microscopy, freeze-fracture and scanning electron microscopy. Gap (communicating) junctions which were observed between mesenchymal cells of all limb bud regions were distributed between apposed cell bodies, points of contact between cell processes and other cell bodies, and between contacting tips of slender cell projections. In addition particularly in the the subridge region, filopodia were observed to extend through the intercellular matrix to contact other cells several micrometers distant. The observations reported in this paper show that mesodermal cells throughout the limb have the structural capability for electrotonic and metabolic coupling during a critical period of morphogenesis in the avian limb. Whether intercellular signals which are thought to be transmitted through gap junctions are active in normal limb development remains to be investigated.

Animals↗

Mesodermalization of amphibian gastrula ectoderm in transfilter experiments.

Transfilter experiments were carried out with competent gastrula ectoderm of Triturus vulgaris and Ambystoma mexicanum as the reacting materials and blastoporal lip, alcohol-treated bone marrow and archenteron roof as the inducing materials, It was shown that a mesodermal inducing substance is able to penetrate Nuclepore filters of different pore sizes (3.0, 1.0, 0.4, 0.2 micrometer). The degree of mesodermal differentiations depends on the reacting materials, the inductors and the filters which were used in the transfilter experiments.

Ambystoma↗

[Cytochemical localization of an important adenyl-cyclase activity in Anuran amphibian embryo chordo-mesoderm by electron microscopy; its possible relations with the migration of primordial germ cells (author's transl)].

Chordo-mesoderm of Anura Amphibia embryos was investigated by electron microscopy and a cytochemical method was used to study the localization of adenyl-cyclase activity, in this part of embryo. It seem that, in Anura, at a given stage of development, there is some adenyl-cyclase activity, and, consequently, some liberation of AMPc. Pleurodeles waltlii (Urodela Amphibia) appears to be devoid of this adenyl-cyclase activity. AMPc could be the substance provided by chordo-mesoderm attracting germinal cells.

Adenylyl Cyclases↗

Mesodermal defect in late phase of gastrulation by a targeted mutation of focal adhesion kinase, FAK.

FAK is a unique non-receptor protein tyrosine kinase that was found in cellular focal adhesions. An increasing number of in vitro observations has suggested that FAK mediates signaling through integrins brought about by interactions with extracellular matrix (ECM). It is highly tyrosine-phosphorylated in v-src-transformed cells and during embryogenesis. To clarify the function of FAK in cell-ECM interactions, embryonic phenotype of its mutant was analysed. FAK-deficient embryos could implant and initiate gastrulation normally, but showed abnormalities in subsequent development. The abnormalities were characterized as a general deficiency in mesoderm, and the phenotype was quite similar to that caused by fibronectin-deficiency. The results suggest that FAK mediates fibronectin-integrin interactions uniquely at this stage of development, thereby playing an essential role in development of mesodermal cell lineages.

Animals↗

An amphioxus Pax gene, AmphiPax-1, expressed in embryonic endoderm, but not in mesoderm: implications for the evolution of class I paired box genes.

Class I paired box genes are widely distributed through the animal phyla but only fruitfly Pox meso and vertebrate Pax-1 and Pax-9 have been adequately characterized. These vertebrate genes have several developmental functions, but their role in patterning the axial skeleton has received the most attention. Because axial skeletons appear after the origin of the vertebrates, special interest attaches to the possible functions of the precursors of Pax-1 and Pax-9 in the invertebrate ancestor of the vertebrates. As a proxy for this ancestor, we studied amphioxus, which is widely thought to be the closest living invertebrate relative of the vertebrates. A cDNA library from developing amphioxus yielded an unequivocal class I paired box gene, AmphiPax-1, that is 2.5 kb long. The gene encodes a 337 amino acid protein that includes a paired domain in which the amino acids are 92% identical to the paired domain amino acids of mouse and human Pax-1 and Pax-9. In situ hybridization detects AmphiPax-1 expression only in the endoderm of the developing pharynx; within this tissue, expression becomes strikingly down-regulated in regions that will fuse with the overlying ectoderm to form gill slits. No transcripts of AmphiPax-1 ever become detectable in any mesodermal structures. We think it likely that, during animal evolution, class I paired box genes originally functioned in endoderm development and were only later co-opted for other roles in mesoderm development; however, other scenarios cannot be ruled out until homologues of these genes are studied in more invertebrate phyla and in the lower vertebrates.

Amino Acid Sequence↗