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PubMed2000

LIF transduces contradictory signals on capillary outgrowth through induction of stat3 and (P41/43)MAP kinase.

The signaling pathways regulating blood vessel growth and development are not well understood. In the present report, an in vitro model was used to identify signaling pathways regulating capillary formation in embryonic endothelial cells. Basic fibroblast growth factor (bFGF) plus leukemia inhibitory factor (LIF) optimally stimulate the formation of capillary-like structures of the embryonic endothelial cell line IEM. LIF stimulation of IEM cells leads to activation of the Stat3 as well as the (P41/43)mitogen-activated protein kinase ((P41/43)MAPK) cascade, while bFGF does not activate Stat3 but does induce the (P41/43)MAPK cascade. Inhibition of Stat3 DNA-binding activity by expression of a dominant inhibitory Stat3 mutant increases the capillary outgrowth of the IEM cells induced by LIF. Increased Stat3 activity by overexpression of the wild-type Stat3 greatly reduced capillary outgrowth. In contrast, inhibition of the (P41/43)MAPK cascade using a MEK-1 inhibitor dramatically inhibits the LIF-induced capillary outgrowth. Moreover, the increased formation of capillary-like structures of the IEM cells mediated by Stat3 inhibition does not overcome the requirement for activation of the (P41/43)MAPK pathway for capillary outgrowth. Stat3 activity correlates with the LIF-induced expression of the negative feedback regulators of the Janus (JAK) family of tyrosine kinases, SOCS-1 and SOCS-3. These results provide evidence that Stat3 acts as a negative regulator of capillary outgrowth, possibly by increasing SOCS-1 or SOCS-3 expression. The contradictory signals stimulated by LIF could be necessary to control the intensity of the response leading to capillary outgrowth in vivo.

Capillaries↗
PubMed1988

Structural analysis of the mitotic cycle in pre-gastrula Xenopus embryos.

The long-known phenomenon of karyomere (chromosome vesicle) formation at early telophase of the nuclear cycle during early embryogenesis of a wide range of organisms including amphibians (Rubaschkin 1905; for review, see Richards 1917) was investigated in the early cleavage cycles of Xenopus laevis embryos before the mid blastula transition. Embryos were fixed and Epon embedded at successive time intervals and consecutive thick (3 micron) and ultrathin sections cut. Using conventional light microscopy at low magnification as well as phase and/or interference contrast video microscopy at high magnification, a substantial amount of information could be obtained from the analysis of optical sections in thick-sectioned material. In addition, details of the ultrastructural organization could be analysed from corresponding ultrathin sections by electron microscopy. The light microscopic analysis of serial thick sections allowed precise determination of the arrangement and sizes of telophase karyomere structures during the embryonic nuclear division cycle. It was found that small, widely spaced 1st order karyomeres fuse to larger (2nd order) karyomeres which then progressively exhibit lateral fusion of neighbouring karyomeres. The final coalescence of adjacent karyomeres marks the onset of the reorganization of the typical interphase nuclear structure. The data are discussed with regard to the occurrence of karyomeres during the embryonic nuclear cycle of arthropods, dipteran insects, and echinoderms as well as recent progress in the use of Xenopus egg extracts for in vitro assembly of nuclear structures around protein-free DNA.

Animals↗
PubMed1976

[Certain structural details of developing in vitro human embryonal brain explants studied with the aid of fluorescence and scanning electron microscopy].

The bundles growing away from the explants were mainly formed by neuroblast processes. It has been shown that the degree of differentiation of neuroblasts depends on "the age" of embryonal brain. The bundles of neuroblasts are accompained by slightly differentiated cells, having their own processes descending to the surface of the lining. These processes attached to the surface of a bundle cover and have no connection with the inner structures of the bundle. The use of a scanning electron microscopy allows to detect some new details of the behaviour of nerve cells in vitro.

Brain↗
PubMed1996

Topography of the "vitreous structures" (tractus; membranelles) with respect to the layers of embryonal blood vessels.

BACKGROUND: It is believed that the vascular or primary vitreous, with the exception of the later Cloquet's canal, gradually disappears and is substituted by the avascular or secondary vitreous. It is known that it is possible topographically to objectify mats of fibril concentrations (membranelles or tractus) of stronger light reflection inside the adult vitreous. These concentrations open up in the shape of a funnel from the papilla or Cloquet's canal towards the front of the vitreous. METHODS: This was a light microscopy investigation on human eyes between the embryonal stage of 3.2 cm and the fetal stage of 12.5 cm and eyes 8 months and 2 years old with persistent vessels of the vitreous. RESULTS: The investigation showed that at the embryonal stage, the vitreous body is threaded from behind by branches (mats) of the hyaloid artery and from in front by vessels (mats) that go over the rim of the optic cup, i.e., the later vitreous base. Vitreous structures, in the form of horsetail-shaped fibril concentrations, could already be observed histologically in the fetal stage with the disappearance of embryonal blood vessels. These structures begin in the vitreous base, go into the vitreous, run parallel to the retina, and then go to the back of the vitreous and towards the lens. The physiological mats of vitreal fibril concentrations (membranelles or tractus) and the pathological branches of the persistent hyaloid artery, topographically correspond to the mats of the obliterated embryonal blood vessels of the vitreous. These mats grow in relation to the bulbous growth. CONCLUSIONS: In these investigations an attempt has been made to clarify the question of which embryonal blood vessels. which embryonal and fetal lengths, which different physiological tractus of the vitreous body and which different pathological features of the persistent hyaloid artery correspond.

Arteries↗
PubMed1982

Cloning, partial sequencing, and expression of glyceraldehyde-3-phosphate dehydrogenase gene in chick embryonic heart muscle cells.

Two recombinant plasmids containing structural gene sequences of chick embryonic heart glyceraldehyde-3-phosphate dehydrogenase (GAP dehydrogenase) were constructed and characterized. The plasmids pGAP 30 and pGAP 36 have inserts of 1200 and 950 base pairs, respectively. The identity of the clones was established by hybrid-arrested and hybrid-selection translation assays, and by immunoprecipitation of hybrid-selected translation product with GaP dehydrogenase antiserum. Hybridization of labeled pGAP 30 DNA to size-fractionated chick heart poly(A) RNA occurred at the region on the gel corresponding to the mobility of GAP dehydrogenase mRNA. Base sequence analysis of plasmid pGAP 30 and the comparison of the amino acid sequence derived from it with that of pig muscle GAP dehydrogenase revealed that the amino acid sequence of GAP dehydrogenase is strictly conserved between the chick and pig muscle tissues. Expression of GAP dehydrogenase mRNA in developing chick heart cells in cultures was monitored by in situ hybridization. The GAP dehydrogenase mRNA was present in 5-h-old dividing myoblasts, in contrast to mRNAs specific for contractile proteins, which appear late in myoblast development paralleling morphogenetic differentiation of myoblasts into myocytes (Jakowlew, S. B., Khandekar, P., Datta, K., Narula, S. K., Arnold, H. H., and Siddiqui, M. A. Q. (1982) J. Mol. Biol. 156, 673-682).

Amino Acid Sequence↗
PubMed1985

SEM-dissection of a human embryo derived from an ectopic pregnancy.

A 6.5-week-old human embryo with an approximate crown-rump length of 13.2 mm was obtained from a tubal pregnancy. Two hours before surgical removal, the embryo was imaged with real time ultrasound and was noted to have rhythmic cardiac motion. Subsequent to surgical removal, the embryo was dissected free from the placenta and prepared for routine scanning electron microscopic (SEM) studies. Progressive stages of dissection with microsurgical instruments followed by SEM photography elucidated the three-dimensional aspects of embryonic development of many structures, including the lens placode, tongue bud, Rathke's pouch, atrial and ventricular foramina, primitive intestinal loop and undifferentiated external genitalia. Almost certainly, such clear views of dissected structures can contribute to our understanding of human embryonic development.

Dissection↗
PubMed2001

Associated arteriovenous malformation of the orbit and brain: a case of Wyburn-Mason syndrome without retinal involvement. Case report.

Wyburn-Mason syndrome is a rare condition associated with multiple cerebral arteriovenous malformations. The disease, also called retinoencephalofacial angiomatosis, includes lesions of the retina, brain, and skin. This disorder stems from a vascular dysgenesis of the embryological anterior plexus early in the gestational period when the primitive vascular mesoderm is shared by the involved structures. The timing of the insult to the embryonic tissue determines which structures are affected. Extensions of the lesions vary widely but cutaneous lesions are unusual. Among reports in the literature, only three cases appear to have manifested without retinal involvement. The authors report the fourth case of Wyburn-Mason syndrome in which there was no retinal involvement and the first to involve neither the retina nor the face.

Cerebrovascular Circulation↗
PubMed1987

Cell death in the embryonic brain of Gallotia galloti (Reptilia; Lacertidae): a structural and ultrastructural study.

In the striatum, thalamus and cerebellum of a Lacertid reptile, we have found three types of cellular death during embryonic development, both at the light and electron microscopic level. The first affects the undifferentiated neuro-epithelial cells and is commonest during the early stages (E. 32-E. 36). The second corresponds to the type of 'nuclear' death described in the bibliography and reaches a maximum in the middle embryonic period (E. 37-E. 39); nevertheless important variations were observed in different zones. The third is the same as the 'cytoplasmic' death type and appears in the perinatal stages. Phagocytosis involved in the elimination of dead cells is of two types. One is associated with early death and is carried out by undifferentiated neuro-epithelial cells. The other is carried out by microglial cells which appear around Stage 37. Much cellular debris was observed in the intermediate zone and this was associated with the second type of phagocytosis. In both cases lipid production was associated with the degenerative process. Comparison of the temporal cellular death pattern with synaptogenesis, gliogenesis and maturation of neuronal processes is consistent with the view that the various types of cellular death found by us had different causes.

Animals↗
PubMed1986

Somitic-vertebral correlation and vertebral levels in the human embryo.

Somitic and vertebral interrelationships and levels were studied in 84 human embryos of stages 9-23 (3-8 postovulatory weeks). The first four somites are occipital, the occipitocervical junction is at somites 4/5, and eight somites are involved in the cervical region: X, Y, Z, and C. 3-7. By stage 17 the total number of occipitovertebral "units," namely 38 or 39, is attained. Resegmentation (Neugliederung) of sclerotomes is not supported. A new scheme of somitic/vertebral correlation is proposed in which somites and centra are in register. Differential growth of the regions of the vertebral column was calculated, and it was found that the percentages of the total column occupied by the various regions vary from one stage to another. The cervical and coccygeal regions decrease, the thoracic and lumbar regions increase, and the sacral region remains more or less constant during embryonic development. The following structures descend with reference to the vertebral column during the embryonic period proper: roots of lower limbs, thyroid gland and thymus, tracheal bifurcation, lungs, heart, diaphragm, abdominal arteries, mesonephroi, and suprarenal glands. The gonads may descend slightly. The scapulae and the separation point between the trachea and the esophagus remain at a fairly constant level. The metanephroi ascend. The migration of many of these structures (e.g., the heart, diaphragm, and metanephroi) is much more marked in the embryonic period than later although it continues during the fetal and postnatal periods. The conus medullaris ascends during the fetal period. Anomalies of migration that affect such organs as the thyroid gland, gonads, and metanephroi are discussed.

Embryo, Mammalian↗
PubMed2003

A new view of patterning domains in the vertebrate mesoderm.

The musculoskeletal system of vertebrates is derived from the embryonic mesoderm. Its structures are categorized as epaxial or hypaxial based on their adult position and innervation. The epaxial/hypaxial terminology is also used to describe regions of the embryonic somites based on fate mapping of somitic derivatives. However, the adult, functional distinctions are not fully consistent with the changing embryonic environments of mesodermal populations during morphogenesis, and the traditional terminology loses accuracy when used to describe certain mutant phenotypes. Here we describe a new terminology naming two mesodermal environments defined by the lineage of the included cells. We discuss how mutant phenotypes may be better explained by consideration of the embryonic context in which genes take their effect and argue that the recognition of these embryonic territories clarifies description and discussion of the morphogenesis and patterning of the musculoskeletal system.

Animals↗
PubMed2000

Histogenesis and morphology of the flexor tendon pulley system in the human embryonic hand.

The number, position, structural and ultrastructural features of the flexor tendon pulley system in six human embryonic hands, aged from 6 to 12 weeks, were studied by light and electron microscope. The pulley system can be recognized from the ninth week; later, at 12 weeks, the structures are easily identified around the flexor tendon in positions closely correlated to those found during post-natal growth and in the adult hand. Structurally and ultrastructurally the pulleys are not simply thickened portions of the sheath. They are formed by three layers: an inner layer, one or two cells thick, probably representing a parietal synovial tendon sheath; a middle layer formed by collagen bundles and fibroblasts whose direction is mainly perpendicular to the underlying phalanx; and an outermost layer consisting of mesenchymal tissue with numerous vessels which extends dorsally in an identical layer, forming a ring that includes flexor and extensor tendons and the cartilaginous model of the phalanx. The pulley does not have a semicircular shape but a much more complicated one, owing to the middle layer which in part runs dorsally and in part ventrally, under the flexor tendons.

Gestational Age↗
PubMed2004

Intermediate filaments are required for C. elegans epidermal elongation.

Cytoplasmic intermediate filaments (cIFs) are thought to provide mechanical strength to vertebrate cells; however, their function in invertebrates has been largely unexplored. The Caenorhabditis elegans genome encodes multiple cIFs. The C. elegans ifb-1 locus encodes two cIF isoforms, IFB-1A and IFB-1B, that differ in their head domains. We show that both IFB-1 isoforms are expressed in epidermal cells, within which they are localized to muscle-epidermal attachment structures. Reduction in IFB-1A function by mutation or RNA interference (RNAi) causes epidermal fragility, abnormal epidermal morphogenesis, and muscle detachment, consistent with IFB-1A providing mechanical strength to epidermal attachment structures. Reduction in IFB-1B function causes morphogenetic defects and defective outgrowth of the excretory cell. Reduction in function of both IFB-1 isoforms results in embryonic arrest due to muscle detachment and failure in epidermal cell elongation at the 2-fold stage. Two other cIFs, IFA-2 and IFA-3, are expressed in epidermal cells. We show that loss of function in IFA-3 results in defects in morphogenesis indistinguishable from those of embryos lacking ifb-1. In contrast, IFA-2 is not required for embryonic morphogenesis. Our data indicate that IFB-1 and IFA-3 are likely the major cIF isoforms in embryonic epidermal attachment structures.

Animals↗
PubMed2004

Molecular cloning and developmental expression of corticotropin-releasing factor in the chicken.

We have characterized the structure of the chicken corticotropin-releasing factor (CRF) gene through cDNA cloning and genomic sequence analysis, and we analyzed the expression of CRF mRNA and peptide in the diencephalon of the chick throughout embryonic development. The structure of the chicken CRF gene is similar to other vertebrate CRF genes and contains two exons and a single intron. The primary structure of the mature chicken CRF peptide is identical to human and rat CRF. This is the first archosaurian CRF gene to be characterized. We used RIAs to analyze CRF peptide content in the diencephalon and the median eminence and plasma corticosterone during the last week of embryonic development. We also developed a semiquantitative RT-PCR method to analyze the expression of CRF mRNA during the same period. CRF peptide content in the diencephalon increased, whereas peptide content in the ME decreased just before hatching, suggesting that release and biosynthesis are coupled. Plasma corticosterone concentration significantly increased between embryonic d 20 and the first day post hatch. By contrast, CRF mRNA levels in the diencephalon decreased just before hatching. Changes in CRF production just before hatching may be causally related to the regulation of the thyroid and interrenal axes at this stage of chicken development.

Aging↗
PubMed1995

Developmental toxicity and structure-activity relationships of chlorophenols using human embryonic palatal mesenchymal cells.

The chlorophenols (CPs) comprise a major class of widely distributed and frequently occurring environmental contaminants. Previous studies have demonstrated the adverse effects of CPs on embryonic and fetal development. HEPM (human embryonic palatal mesenchymal) and MOT (mouse ovarian tumor) cell lines have been utilized in complementary bioassays for the detection of teratogens, but not the CPs. In this study, our objectives were 2-fold: (1) to determine if the HEPM assay could be used to complement other bioassay systems of nonhuman origin, i.e., Hydra attenuata (HA) and rat whole embryo culture (WEC), in the evaluation of the developmental toxicity of CPs, and (2) to delineate the ability of the HEPM assay to evaluate structure-activity relationships of pentachlorophenol (C5P), 2,3,4,5-tetrachlorophenol (C4P), 2,3,5-trichlorophenol (C3P), 3,5-dichlorophenol (C2P), 4-monochlorophenol (CP), phenol, and CP derivatives (i.e., acetates, sodium phenates and anisoles). HEPM cells were seeded into each well of a 24-well plate and cultivated for 24 h. The medium was replaced with fresh medium containing various concentrations of test chemicals dissolved in dimethyl sulfoxide (DMSO, 0.1%). After culturing for 72 h, the medium was removed, cells were trypsinized, and cell number determined. The HEPM cell growth inhibition assay demonstrated a linear relationship between the IC50 values of the CPs and degree of chlorine substitution. The IC50 values of C5P, C4P, C3P, C2P, CP, and phenol were 18.8, 21.5, 27.5, 63.0, 150.0 and 470.0 microM, respectively. A clear structure-activity relationship was observed between toxicity of CPs and the degree of chlorine substitution. The rank order of CP toxicity from the HEPM assay (i.e., C5P > C4P > C3P > C2P > CP > phenol) is in excellent agreement with previous in vitro and in vivo studies. However, contrary to published reports, the HEPM assay predicted that all CPs were teratogenic (false positives). These findings suggest that the HEPM cell growth inhibition bioassay may be useful to discriminate between subtle differences in structure-activity and, in combination with other bioassays, might facilitate the rapid detection and prioritization of diverse cytotoxins, including various developmental toxicants. Importantly, conclusions about the teratogenicity of a test chemical (via HEPM testing) should be approached with caution and confirmed with other teratogen-sensitive systems.

Animals↗
PubMed1999

[The characteristics of actin cytoskeleton structure and its rearrangements by extracellular matrix proteins in normal, immortalized and transformed rat fibroblasts].

Comparative analysis of actin cytoskeleton structure in rat embryonic fibroblasts, E1A-immortalized and E1A + cHa-ras-transformed cells has been carried out. A decrease in adhesiveness and the rate of changes in actin cytoskeleton structures was shown to correlate with the level of morphological transformation of cells. E1A + cHa-ras-transformants show the lowest adhesiveness and complete disorganization of actin structures. Cultivation on serum-free media promoted disassembling of actin cytoskeleton structures in a small part of normal fibroblast population, only in a few immortalized cells, but exerted no influence on transformed cells. The influence of immobilized extracellular matrix proteins fibronectin, laminin and collagens type I and III on actin cytoskeleton structure in normal, immortalized and transformed fibroblasts was studied. Transformed cells spread on fibronectin completely restored highly organized actin structures, displayed a lot of stress fibers and focal contacts. The use of laminin revealed differences in locomotion between normal and transformed cells. Normal, immortalized and transformed fibroblasts spread on fibronectin and laminin demonstrate some peculiarities in actin cytoskeleton structures as a result of specificity of ligand-receptor interaction. Cells spread on fibronectin have polygonal shapes, many stress fibers and focal contacts, whereas cells spread on laminin are highly polarized and develop broad lamellae filled with actin microfilament meshwork. Collagens type I and III can affect adhesive properties and actin cytoskeleton structure in all cell lines studied only slightly, in comparison with fibronectin and laminin.

Actins↗
PubMed1998

Blocking N-cadherin function disrupts the epithelial structure of differentiating neural tissue in the embryonic chicken brain.

The cell adhesion molecule N-cadherin is ubiquitously expressed in the early neuroepithelium, with strongest expression in the ependymal lining. We blocked the function of N-cadherin during early chicken brain development by injecting antibodies against N-cadherin into the tectal ventricle of embryos at 4-5 d of incubation [embryonic day 4 (E4)-E5]. N-cadherin blockage results in massive morphological changes in restricted brain regions. At approximately E6, these changes consist of invaginations of pieces of the ependymal lining and the formation of neuroepithelial rosettes. The rosettes are composed of central fragments of ependymal lining, surrounded by an inner ventricular layer and an outer mantle layer. Radial glia processes are radially arranged around the ependymal centers of the rosettes. The normal layering of the neural tissue is thus preserved, but its coherent epithelial structure is disrupted. The observed morphological changes are restricted to specific brain regions such as the tectum and the dorsal thalamus, whereas the ventral thalamus and the pretectum are almost undisturbed. At E10-E11, analysis of late effects of N-cadherin blockage reveals that in the dorsal thalamus, gray matter is fragmented and disorganized; in the tectum, additional layers have formed at the ventricular surface. Together, these results indicate that N-cadherin function is required for the maintenance of a coherent sheet of neuroepithelium in specific brain regions. Disruption of this sheet results in an abnormal morphogenesis of brain gray matter.

Animals↗
PubMed1980

[Protein synthesis in different rabbit embryo tissues in the presence of utero-placental ischemia].

Resistance of different elements of the hemodynamic functional "mother--placenta--fetus" system to the damaging effect (ligation of a half of preplacental vessels, performed on the 8th day of pregnancy) was studied and compared in experiments on female rabbits on the 12th day of pregnancy. It appeared that under these conditions the extent of growth retardation in different embryonic and placental structures is dissimilar. The processes of cytoplasmic protein synthesis in heart tissues are more resistant to the damaging effect than in bodily tissues. The delayed hemoglobin accumulation in the embryonic organism was not so pronounced as retardation of protein synthesis in the body.

Animals↗