Purification of a RNA-binding protein from rat liver. Identification as ferritin L chain and determination of the RNA/protein binding characteristics.

In cultured rat hepatocytes the degradation of phosphoenolpyruvate carboxykinase mRNA might be regulated by protein(s), which by binding to the mRNA alter its stability. The 3'-untranslated region of phosphoenolpyruvate carboxykinase mRNA as a potential target was used to select RNA-binding protein(s) from rat liver by the use of gel retardation assays. A cytosolic protein was isolated, which bound to the phosphoenolpyruvate carboxykinase mRNA 3'-untranslated region and other in vitro synthesized RNAs.

VEGF and VEGF-C: specific induction of angiogenesis and lymphangiogenesis in the differentiated avian chorioallantoic membrane.

The lymphangiogenic potency of endothelial growth factors has not been studied to date. This is partially due to the lack of in vivo lymphangiogenesis assays. We have studied the lymphatics of differentiated avian chorioallantoic membrane (CAM) using microinjection of Mercox resin, semi- and ultrathin sectioning, immunohistochemical detection of fibronectin and alpha-smooth muscle actin, and in situ hybridization with VEGFR-2 and VEGFR-3 probes.

Mechanism of the impairment of the glucagon-stimulated phosphoenolpyruvate carboxykinase gene expression by interleukin-6 in rat hepatocytes: inhibition of the increase in cyclic 3',5' adenosine monophosphate and the downstream cyclic 3',5' adenosine monophosphate action.

In cultured rat hepatocytes, the gluconeogenic key enzyme, phosphoenolpyruvate carboxykinase (PCK), is induced by glucagon via elevation of cyclic 3',5' adenosine monophosphate (cAMP). The proinflammatory cytokine, interleukin-6 (IL-6), which in the liver together with IL-1beta and tumor necrosis factor alpha triggers the acute-phase response, had been shown to attenuate the glucagon-induced increase in PCK gene transcription, messenger (mRNA) levels, and enzyme activity. The molecular mechanism of this inhibition was investigated in the present study.

Regulation of paraxis expression and somite formation by ectoderm- and neural tube-derived signals.

During vertebrate embryogenesis, the paraxial mesoderm becomes segmented into somites, which form as paired epithelial spheres with a periodicity that reflects the segmental organization of the embryo. As a somite matures, the ventral region gives rise to a mesenchymal cell population, the sclerotome, that forms the axial skeleton. The dorsal region of the somite remains epithelial and is called dermomyotome.

The fate of the first avian somite.

We have studied the derivatives of the first somite using the quail-chick marking technique. After transplantation of the somite, the chick embryos were reincubated for periods ranging from 4 h to 11 days. Coronal and sagittal sections of the embryos were prepared for parallel staining with Feulgen-reaction, anti-quail antibody, anti-desmin antibody and QH-1 antibody.

Expression of the avian VEGF receptor homologues Quek1 and Quek2 in blood-vascular and lymphatic endothelial and non-endothelial cells during quail embryonic development.

We have studied the expression of Quek1 and Quek2 (VEGFR-2 and VEGFR-3, respectively) in quail embryos from day 2 to day 16 by in situ hybridization with digoxigenin-labelled riboprobes on whole-mounts and paraffin sections. Parallel sections were also stained with the QH1 antibody to detect all endothelial cells and with an antibody against alpha-smooth-muscle-actin to reveal the media of blood vessels. Quek1/VEGFR-2 is a marker of blood-vascular and lymphatic endothelial cells throughout development.

Insertion of musculus tibialis posterior into musculus peroneus (fibularis) longus.

We report on a muscular variation in the human foot. The tendon of the tibialis posterior muscle was attached to the peroneus longus tendon by means of a trapezoid tendinous connection. This variation was observed only in the left foot.

Location and growth of epaxial myotome precursor cells.

The skeletal muscle progenitor cells of the vertebrate body originate in the dermomyotome epithelium of the embryonic somites. To precisely locate myotome precursor cells, fluorescent vital dyes were iontophoretically injected at specific sites in the dermomyotome in ovo and the fates of dye-labeled cells monitored by confocal microscopy. Dye-labeled myotome myofibers were generated from cells injected along the entire medial boundary and the medial portion of the cranial boundary of the dermomyotome, regions in close proximity to the dorsal region of the neural tube where myogenic-inducing factors are thought to be produced.

Mesoderm-derived cells proliferate in the embryonic central nervous system: confocal microscopy and three-dimensional visualization.

In the chick and quail embryo, two cell populations migrate into the neural tube from the surrounding mesodermal tissues during the fourth day of incubation: individual cells which represent macrophages, and endothelial cells which remain continuous with the extraneural vessels. We report here on the proliferative capacity of these mesoderm-derived cells. A double-immunofluorescence protocol for two monoclonal antibodies of subtype IgG1, the endothelial cell/macrophage marker QH1, and the S-phase marker bromodeoxyuridine, was developed.

Identification and characterization of differentiation-dependent Schwann cell surface antigens by novel monoclonal antibodies: introduction of a marker common to the non-myelin-forming phenotype.

In an attempt to identify and characterize novel Schwann cell surface molecules with putative functions during development, maintenance, and regeneration of the peripheral nervous system (PNS), we have produced monoclonal antibodies against viable neonatal rat Schwann cells. Using a sensitive live cell ELISA protocol, three monoclonal antibodies reactive with cultured Schwann cells, designated 27B10, 26F2, and 27C7 were isolated. The 27B10 and 26F2 antibodies specifically labelled forskolin-stimulated secondary Schwann cells in vitro as determined by live cell ELISA implying that the expression of the antigens in situ is regulated by axonal contact.

On the bifurcation of blood vessels--Wilhelm Roux's doctoral thesis (Jena 1878)--a seminal work for biophysical modelling in developmental biology.

Wilhelm Roux's doctoral thesis described the relationship between the angle and diameter of bifurcating blood vessels. We have re-read this work in the light of biophysics and developmental biology and found two remarkable aspects hidden among a multitude of observations, rules and exceptions to these rules. First, the author identified the major determinants involved in vascular development; genetics, cybernetics, and mechanics; moreover, he knew that he could not deal with the genetic and regulatory aspects, and could hardly treat the mechanical part adequately.

Diastematomyelia and spina bifida can be caused by the intraspinal grafting of somites in early avian embryos.

Objective: In this experimental study, an embryological model was created to reproduce diastematomyelia and spina bifida and to investigate new aspects of the origin of spinal cord malformations.

Methods: A somite was implanted from a donor quail embryo into the neural tube of a 2-day-old chick embryo. The somite was chosen because the septum that characteristically separates the two hemicords consists exclusively of mesodermal derivatives.

Impairment by interleukin 1 beta and tumour necrosis factor alpha of the glucagon-induced increase in phosphoenolpyruvate carboxykinase gene expression and gluconeogenesis in cultured rat hepatocytes.

The influence of the inflammatory mediators interleukin 1 beta (IL1 beta) and tumour necrosis factor alpha (TNF alpha) on the glucagon-induced expression of phosphoenolpyruvate carboxykinase (PCK) and on glucose formation via gluconeogenesis was investigated in cultured rat hepatocytes. Gene expression was monitored by determination of mRNA levels and of enzyme activity. Glucose formation was estimated with newly synthesized radioactive glucose derived from a radiolabelled lactate precursor.

Scatter factor/hepatocyte growth factor (SF/HGF) induces emigration of myogenic cells at interlimb level in vivo.

The initiating event in the migration of myogenic cells to the limb buds is an epitheliomesenchymal transformation of cells located at the lateral edge of the dermomyotome. Recently, a targeted mutation of c-met in mice demonstrated an essential role of this tyrosine kinase receptor and its ligand, scatter factor/hepatocyte growth factor (SF/HGF), in the migration of myogenic cells to the limb buds. Here, we show that ectopic application of exogenous SF/HGF induces emigration of Pax-3-positive myogenic cells into the lateral plate mesoderm.

Participation of individual brachial somites in skeletal muscles of the avian distal wing.

In this paper we investigate the somitic origin of the individual muscles of the forearm and hand using quail-chick chimeras. Our results show that only somites 16-21 give rise to wing muscle, but they take part in muscle formation to different extents. Somite 21 does not always participate in the formation of muscle of the forearm and hand.

Expression of avian Pax1 and Pax9 is intrinsically regulated in the pharyngeal endoderm, but depends on environmental influences in the paraxial mesoderm.

Pax1 and Pax9 represent a subfamily of paired-box-containing genes. In vertebrates, Pax1 and Pax9 transcripts have been found specifically in mesodermal tissues and the pharyngeal endoderm. Pax1 expression in the sclerotomes has been shown to be indispensable for proper formation of the axial skeleton, but expression of Pax1 in the endoderm has not been studied in detail.

The expression and regulation of follistatin and a follistatin-like gene during avian somite compartmentalization and myogenesis.

We report on the normal and experimentally altered expression of two structurally related genes, Follistatin and Follistatin-like (Flik), in the somites of avian embryos. In normal chick embryos, Follistatin expression can first be seen in the cells of the dorsolateral somite quarter. During somite maturation, the cells of the dorsomedial quarter also express this gene.

N-cadherin is involved in myoblast migration and muscle differentiation in the avian limb bud.

Limb muscle formation involves invasion of the limb bud mesoderm by myogenic precursor cells from the dermomyotomes at limb bud level. Directed cell migration, homing, and differentiation of myogenic cells are controlled by the stationary cells of the limb bud mesoderm. At the level of the extracellular matrix, the molecular basis of migration control has been suggested to be exerted by the distribution of hyaluronan.

Fibroblast growth factor receptor 1 in skeletal and heart muscle cells: expression during early avian development and regulation after notochord transplantation.

Basic fibroblast growth factor (bFGF, FGF-2) mediates several biological functions during embryonic development. With regard to skeletal muscle formation, it has been suggested that FGF-2 is involved in the growth and differentiation of myogenic precursor cells. To identify the FGF-responsive cells we studied the expression of FGF receptor type I (FGFR-1) during early embryonic development of the chick.

VEGF121 induces proliferation of vascular endothelial cells and expression of flk-1 without affecting lymphatic vessels of chorioallantoic membrane.

We have studied the effect of VEGF(121) homodimer and VEGF(121/165) heterodimer on the chorioallantoic membrane (CAM) of 13-day-old chick embryos. The factors were applied in doses of 2-4 micrograms and the effects were evaluated macroscopically after 2 and 3 days. Histological studies were performed on semi- and ultrathin sections.

Molecular cloning, sequencing and expression of the cDNA of the mitochondrial form of phosphoenolpyruvate carboxykinase from human liver.

In human liver, phosphoenolpyruvate carboxykinase (PCK; EC 4.1.1.

Axial structures control laterality in the distribution pattern of endothelial cells.

In the midline of the embryo an invisible barrier exists that keeps endothelial cells from migrating to the contralateral side. Interspecific grafting experiments between chick and quail were carried out in order to investigate the role of the axial structures in maintaining this barrier. The quail endothelial cells of the graft were therefore stained with QH1 antibody.

Embryonic angiogenesis: a review.

Supply with nutrients is essential from early embryonic stages onwards. Therefore, circulatory organs form the first functioning organ system. With the exception of the heart, this system is at first formed by only one cell type, the endothelial cell.