bluuebunny/crossref_metadata_2025_split huggingface dataset:
Stability of RNA in developing Xenopus embryos and identification of a destabilizing sequence in TFIIIA messenger RNA
Authors: Harland Richard , Misher Lynda 
Published: April 2021

Abstract
Synthetic capped RNA transcripts injected into fertilized eggs of Xenopus laevis have a half-life of 3 –4 h. Addition of a long (∼200 nucleotide) poly(A) tail increases the half-life to 6 –8 h which approaches the half-life of natural polyadenylated globin RNA injected into embryos. Since exonucleolytic action alone could account for the degradation of RNA, we tested whether circular RNA is stable after injection and find that circles are exceptionally stable (half-life greater than 40 h). After the midblastula transition, polyadenylated chloramphenicol transferase (CAT) mRNAs transcribed from injected plasmids have a half-life of 2 ·5 h. Insertion of a 1000 nucleotide 3′ untranslated region from the Xhox-36 gene into the transcripts does not affect the half-life. In contrast to the finding that internal sequences do not affect stability, we find that sequences from the TFIIIA message reduce the half-life of CAT mRNA from 2 ·5 h to less than 30 min. We conclude that most RNAs are degraded exonucleolytically from the 3′ end, but specialized internal sequences can greatly destabilize the RNA, possibly by acting as a site for an endonuclease.
Segmental pattern of development of the hindbrain and spinal cord of the zebrafish embryo
Authors: Hanneman Eric , Trevarrow Bill , Metcalfe Walter K. , Kimmel Charles B. , Westerfield Monte 
Published: April 2021

AbstractIn the ventral hindbrain and spinal cord of zebrafish embryos, the first neurones that can be identified appear as single cells or small clusters of cells, distributed periodically at intervals equal to the length of a somite. In the hindbrain, a series of neuromeres of corresponding length is present, and the earliest neurones are located in the centres of each neuromere. Young neurones within both the hindbrain and spinal cord were identified in live embryos using Nomarski optics, and histochemically by labelling for acetylcholinesterase activity and expression of an antigen recognized by the monoclonal antibody zn-1. Among them are individually identified hindbrain reticulospinal neurones and spinal motoneurones. These obervations suggest that early development in these regions of the CNS reflects a common segmental pattern. Subsequently, as more neurones differentiate, the initially similar patterning of the cells in these two regions diverges. A continuous longitudinal column of developing neurones appears in the spinal cord, whereas an alternating series of large and small clusters of neurones is present in the hindbrain.
The contribution made by a single somite to the vertebral column: experimental evidence in support of resegmentation using the chick–quail chimaera model
Authors: Bagnall K. M. , Higgins S. J. , Sanders E. J. 
Published: April 2021

AbstractThe somitic involvement in the formation of the vertebral column was examined using the chick-quail chimaera model. Single cervical somites from quail donor embryos were transplanted into similarly staged chick host embryos. Following further incubation, serial sections of variously staged embryos were stained with the Feulgen reaction to distinguish the two cell populations.Quail cells were generally located within a delimited region in one half of each of the two adjacent vertebrae, as well as in the intervening disc. The horizontal plane of division through each vertebra passed approximately through the centre of the body and divided the neural arch into rostral and caudal halves through the rostral border of the caudal notch. These results give support to the controversial theory of resegmentation, in which it was suggested that there is an apparent realignment of segmentation between the somite stage and the subsequent vertebral stage of development.
A Dictyostelium prespore-specific gene is transcriptionally repressed by DIF in vitro
Authors: Early A. E. , Williams J. G. 
Published: April 2021

Abstract
One important role of DIF, the stalk cell-specific inducer of Dictyostelium, may be to divert cells from the spore cell pathway of differentiation. The D19 gene encodes an mRNA which is highly enriched in prespore over prestalk cells in the migratory slug. We show, using a mutant defective in DIF accumulation, that the concentration of D19, and several other prespore mRNA sequences, decreases in the presence of exogenous DIF. There is evidence that both transcriptional and post-transcriptional controls operate to regulate expression of these genes. We have performed in vitro nuclear transcription and mRNA halflife analyses, and find that DIF acts at the transcriptional level to repress the accumulation of the D19 mRNA.
Origin of the dorsal surface of the neural tube by progressive delamination of epidermal ectoderm and neuroepithelium: implications for neurulation and neural tube defects
Authors: Martins-Green Manuela 
Published: April 2021

Abstract
Knowledge of the morphogenetic events involved in the development of the dorsal portion of the neural tube is important for understanding neural tube closure, neural crest cell formation and emigration, and the origin of neural tube defects. Here, I characterize the progressive development of the tips of the neural folds during fold elevation in the trunk of mouse and chick embryos and the events leading to formation of the dorsal portion of the neural tube as the epidermal ectoderm (EE) and neuroepithelium (NE) separate from each other. The nature and timing of appearance of collagen IV, laminin and fibronectin were analysed by immunofluorescent and immunogold labelling, and ruthenium red and tannic acid were used to enhance staining for proteoglycans and glycosaminoglycans. As the neural folds elevate, the NE and EE delaminate progressively beginning at the basal surface of the lateral extremes of the neural plate. Nevertheless, the two epithelia remain connected across the zone of delamination by their previously existing basal laminae. In each fold, proteoglycan granules appear at the interface between the NE and EE before delamination begins, and then an (inter-epithelial) space begins to open and propagate dorsally. Other extracellular matrix (ECM) molecules appear within the space a short distance behind its tip and basal lamina deposition begins shortly thereafter. As fusion occurs, the interepithelial spaces of the two folds coalesce and the final separation of the EE from the NE is accomplished. These observations suggest that the previously recognized delay in deposition of ECM and basal lamina on the dorsal portion of the neural tube and on the overlying EE is a direct consequence of the delamination of the two epithelia and the establishment of two new basal surfaces. The observation that the surface of the dorsal third of the neural tube forms by delamination rather than by juxtaposition of previously existing basal surfaces of the two epithelia is discussed in terms of possible implications for models of neurulation and the origin of neural tube defects.
Pepsinogen gene transcription induced in heterologous epithelial—mesenchymal recombinations of chicken endoderms and glandular stomach mesenchyme
Authors: Hayashi Kensuke , Yasugi Sadao , Mizuno Takeo 
Published: April 2021

AbstractProventricular (glandular stomach) mesenchyme of chicken embryos can induce endoderms of some parts of embryonic digestive tract to produce embryonic chicken pepsinogen (ECPg), a marker protein for the differentiation of embryonic proventricular epithelium. In the present study, we investigated the production of ECPg mRNA in the course of epithelial-mesenchymal interactions between endoderms of digestive tract and proventricular mesenchyme. ECPg mRNA was detected by Northern hybridization with ECPg cDNA as a probe. In normal development of the proventriculus, ECPg mRNA was first detected at day 7 of incubation, and it ceased to be produced by day 21. Embryonic esophagus, gizzard and small intestine did not contain ECPg mRNA. When 6-day esophagus, gizzard or proventricular endoderm was associated and cultured with 6-day proventricular mesenchyme, the recombinates formed proventricular-gland-like complex glands and produced ECPg mRNA in almost equal quantity. However, 6-day small intestinal or 3·5-day allantoic endoderm did not produce pepsinogen mRNA under the same conditions, though the recombinates formed complex glands. These results indicate that the proventricular mesenchyme can induce de novo transcription of ECPg gene in esophagus, proventricular and gizzard endoderms, and that ECPg gene in small intestinal and allantoic endoderms fails to react to the inducing signal.
The expression of epidermal antigens inXenopus laevis
Authors: Itoh Keiji , Yamashita Akiko , Kubota Hiroshi Y. 
Published: April 2021

ABSTRACTFive kinds of monoclonal antibodies that are specific for the epidermis of Xenopus embryos were produced. Epidermis-specific antibodies were used to investigate the spatial and temporal expressions of epidermal antigens during embryonic and larval development. The cells that were recognized by the antibodies at the larval stage are as follows: all of the outer epidermal cells and cement gland cells were recognized by the antibody termed XEPI-1, all of the outer and inner epidermal cells, except the cement gland cells, were recognized by XEPI-2 antibody, the large mucus granules and the apical side of the outer epidermal cells, except for the ciliated epidermal cells, were recognized by XEPI-3 antibody, the large mucus granules and basement membrane were recognized by XEPI-4 antibody, and the small mucus granules contained in the outer epidermal cells as well as extracellular matrices were recognized by the antibody termed XEPI-5. All of the epidermal antigens, except XEPI-4, were first detected in the epidermal region of the late gastrula or early neurula. The XEPI-4 antigen was first detected in stage-26 tail-bud embryos. None of these antigens were expressed by the neural tissues at any time during embryonic development. Only the XEPI-2 antigen continued to be expressed after metamorphosis, while the expression of the other antigens disappeared during or before metamorphosis. The specificity of the antibodies allowed us to classify the epidermal cells into four types in early epidermal development. The four types of epidermal cells are (1) the outer epidermal cells that contain small mucus granules, (2) the ciliated epidermal cells, (3) the outer epidermal cells that contain large mucus granules and (4) the inner sensorial cells.
Embryonic expression pattern of TGF beta type-1 RNA suggests both paracrine and autocrine mechanisms of action
Authors: Lehnert Sigrid A. , Akhurst Rosemary J. 
Published: April 2021

ABSTRACT
The tissue distribution of TGF beta form 1 RNA within mouse embryos of 10·5 to 15·5 days gestational age was investigated using in situ hybridization. As predicted from the prevalence of TGF beta-1 protein in adult bone and platelets, the RNA is highly abundant in fetal bone and in fetal liver megakaryocytes. Our data also reveal previously undocumented sites of synthesis for TGF beta-1, namely epithelia overlying those mesenchymal tissues that are known to contain TGF beta protein as detected by immunohistochemical methods (Heine et al. 1987) and in the mesenchymal tissues of certain internal organs. From a combined knowledge of the distributions of the TGF beta polypeptide (Heine et al. 1987) and its mRNA, and a knowledge of the reported biological activities of TGF beta-1, we invoke both paracrine and autocrine mechanisms of action for this growth factor.
Isolation of differentially expressed human cDNA clones: similarities between mouse and human embryonal carcinoma cell differentiation
Authors: Wiles Michael V. 
Published: April 2021

ABSTRACTThe study of early human development is of great importance but has been limited by the lack of suitable reagents. Recently, however, the human embryonal carcinoma (EC) cell line NT2D1 has been isolated. This cell line will differentiate upon exposure to retinoic acid (RA). A cDNA library was constructed from poly(A)+ RNA derived from NT2D1 cells treated with 10−5M-RA for 7 days (ANT2D1 cells). By differential cDNA screening, it was found that 1-12% of ANT2D1 cDNA recombinants screened detected an increase in signal with 32P-cDNAs derived from ANT2D1 as compared with NT2D1.To compare RA-induced differentiation of mouse and human EC cells, the ANT2D1 cDNA library was rescreened with 32P-cDNAs derived from the mouse EC cell line F9 and the result compared with 32P-cDNA derived from F9 differentiated to parietal-endoderm (F9PE)-like cells and visceral-endoderm (F9VE)-like cells. Approximately 1·2% of the ANT2D1 cDNA recombinants detected a differential increase in signal following differentiation of mouse EC cells to F9VE and/or F9PE. Of these homologous regulated sequences, 0·3 % were common to both mouse and human EC cell RA-induced differentiation. Five different cDNA clones were isolated that detect a marked increase (5- to 75-fold) in mRNA abundance following RA-induced differentiation of NT2D1. Of these five clones, three detect homologous mRNAs which also increase in abundance following differentiation of the mouse EC cell line F9 to PE- and/or VE-like cells; the other two clones do not detect sequences in the mouse mRNAs tested. One clone shows homology to SPARC, a gene known to be regulated during mouse embryonic development. While another clone, SO5A, has a limited range of expression, being detected in F9VE and in a human parietal-endodermlike cell, but not in F9PE and a human visceral-endoderm-like cell.This work shows that there are both similarities and differences in mouse and human EC cell differentiation, and these cDNA clones provide some of the first reagents for studying the molecular biology of human development.
Expression of tenascin in response to neural induction in amphibian embryos
Authors: Riou Jean-Francois , Shi De-Li , Chiquet Matthias , Boucaut Jean-Claude 
Published: April 2021

ABSTRACTThe expression of tenascin, a constituent of extracellular matrix (ECM), was studied during the embryonic development of the amphibian Pleurodeles waltl. An antiserum to chick fibroblast tenascin was shown to cross-react with the homologous molecule of the amphibian. Immunostaining of embryo sections with anti-tenascin antiserum revealed that tenascin appears just after the completion of neurulation. At the tailbud stage, tenascin is present in the ECM located at sites of directed cell migration (neural crest cell migration pathways, extension of the pronephretic duct) and mesenchyme condensation (endocardium, aortic arches). The accumulation of tenascin immunoreactivity in the embryonic ECM is correlated with the synthesis of the 220×103A/r polypeptide of the molecule. To provide data on the patterning of tenascin, ectoderm and dorsal blastoporal lip isolated at early gastrula stage were cultured for a period of 3 days. Epidermal vesicles differentiating from isolated ectoderm completely lack tenascin. Conversely, axial mesoderm derivatives present in cultured dorsal blastoporal lip were found to produce tenascin. Neural induction of ectoderm isolated at early gastrula stage was performed in vitro with the dorsal blastoporal lip or concanavalin A. The induced neural tissue was found to accumulate tenascin. Spemann experiments confirmed in vivo that tenascin is expressed by ectodermal cells as a response to neural induction.
Requirement of cell division for muscle actin expression in the primary muscle cell lineage of ascidian embryos
Authors: Jeffery William R. 
Published: April 2021

Abstract
The role of cell division in the expression of muscle actin and its relationship to acetylcholinesterase (AChE) development was examined in cleavage-arrested embryos of the ascidian Styela. Muscle actin expression was detected by two-dimensional gel electrophoresis of radioactively labelled proteins and by in situ hybridization with a cDNA probe, whereas AChE activity was assayed by enzyme histochemistry. In the majority of cases, muscle actin expression was first detected in embryos arrested after the 16-cell stage. Some embryos showed muscle actin expression after arrest at the 8-cell stage, however, muscle actin mRNA did not accumulate in embryos arrested at earlier cleavages. The cells that expressed muscle actin in 8- to 64-cell cleavage-arrested embryos belonged to the primary muscle lineage; secondary muscle cell precursors did not express muscle actin. Zygotic muscle actin mRNA appeared to accumulate with myoplasmic pigment granules in the perinuclear region of cleavage-arrested embryos, suggesting that the myoplasm may have a role in the organization of muscle cells. In contrast to muscle actin, AChE was detected in a small proportion of embryos treated with cytochalasin as early as the 1- or 2-cell stage, and most embryos treated with cytochalasin at later cleavages expressed this enzyme in some of their cells. Most primary muscle lineage cells expressed both muscle actin mRNA and AChE, however, some cells expressed only muscle actin mRNA or AChE. The results suggest that at least three cleavages are required for muscle actin expression and that muscle actin and AChE expression can be uncoupled in cleavage-arrested embryos.
Proliferation and differentiation potential of rat forebrain oligodendroglial progenitors both in vitro and in vivo
Authors: Hardy Rebecca , Reynolds Richard 
Published: April 2021

ABSTRACT
We have followed the development of the O-2A progenitor cell from the neonatal rat forebrain, both in dissociated cell culture and in cryostat sections, using immunocytochemical techniques employing a panel of antibodies that recognise the cells at different stages of their development. This included the monoclonal antibody LBi, which binds to the surface ganglioside GD3 expressed on 0-2A progenitor cells. In secondary cultures enriched for O-2A progenitors maintained in a serum-free chemically defined medium, a large proportion of the cells are primed to differentiate into oligodendroglia and go on to express the oligodendroglial specific surface glycolipid galactocerebroside (GC) and then the myelin proteins CNP and MBP. However, a significant proportion of immature bipolar GD3+ cells remained after 6 days in secondary culture. It appears that not all the O-2A progenitors in our cultures differentiate immediately and some cells remain in an undifferentiated state and divide to replenish progenitor numbers. We have also identified in our cultures a small apolar GD3- cell, which when isolated differentiated into a GD3+ bipolar O-2A progenitor cell. We have termed this cell type a preprogenitor. The differentiation of this cell type into O-2A progenitors may be the source of the immature GD3+ cells present at the later stages of our secondary cultures. The proliferative profile of the cultures was studied using 5’bromo-2-deoxyuridine (BrdU) incorporation as an index of mitosis. Only the immature, bipolar O-2A progenitors were seen to divide at any time in serum-free culture. Neither the more mature multipolar O-2A cells nor the oligodendroglia were seen to divide. The developmental profile of the O-2A cells in the rat forebrain in vivo showed a largely similar progression to that in culture, with a time lag of at least 6 days between GD3 expression and the onset of myelination. BrdU incorporation studies in vivo also showed that the GD3+ progenitor cell is mitotic whereas the GC+-expressing oligodendroglia is not. We have shown that there are several significant alterations in the timing of antigen expression in both O-2A progenitors and oligodendroglia in vitro compared to that seen in vivo.
Origins and patterning of avian outflow tract endocardium
Authors: Noden Drew M. 
Published: April 2021

ABSTRACT
Outflow tract endocardium links the atrioventricular lining, which develops from cardiogenic plate mesoderm, with aortic arches, whose lining forms collectively from splanchnopleuric endothelial channels, local endothelial vesicles, and invasive angioblasts. At two discrete sites, outflow tract endocardial cells participate in morphogenetic events not within the repertoire of neighboring endocardium: they form mesenchymal precursors of endocardial cushions.
The objectives of this research were to document the history of outflow tract endocardium in the avian embryo immediately prior to development of the heart, and to ascertain which, if any, aspects of this history are necessary to acquire cushion-forming potential. Paraxial and lateral mesodermal tissues from between somitomere 3 (midbrain level) and somite 5 were grafted from quail into chick embryos at 3–10 somite stages and, after 2–5 days incubation, survivors were fixed and sectioned. Tissues were stained with the Feulgen reaction to visualize the quail nuclear marker or with antibodies (monoclonal QH1 or polyclonals) that recognize quail but not chick cells. Many quail endothelial cells lose the characteristic nuclear heterochromatin marker, but they retain the species-specific epitope recognized by these antibodies.
Precursors of outflow tract but not atrioventricular endocardium are present in cephalic paraxial and lateral mesoderm, with their greatest concentration at the level of the otic placode. Furthermore, the ventral movement of individual angiogenic cells is a normal antecedent to outflow tract formation. Cardiac myocytes were never derived from grafted head mesoderm. Thus, unlike the atrioventricular regions of the heart, outflow tract endocardial and myocardial precursors do not share a congruent embryonic history.
The results of heterotopic transplantation, in which trunk paraxial or lateral mesoderm was grafted into the head, were identical, including the formation of cushion mesenchyme. This means that cushion positioning and inductive influences must operate locally within the developing heart tubes.
The snail gene required for mesoderm formation in Drosophila is expressed dynamically in derivatives of all three germ layers
Authors: Alberga Audrey , Boulay Jean-Louis , Kempe Elisabeth , Dennefeld Christine , Haenlin Marc 
Published: April 2021

ABSTRACT
The zygotic effect gene snail (sna) encodes a zinc-finger protein required for mesodenn formation in Drosophila embryos. By in situ analysis, sna transcripts are first detected at syncytial blastoderm and persist until very late stages of embryogenesis. Expression of sna is transient and is observed in tissues derived from all three germ layers. Prior to germband elongation, sna RNA accumulation is consistent with its genetically deter mined role in mesoderm formation. Starting at germ band elongation, a second phase of sna expression appears to be initiated, characterized by a highly dynamic accumulation of transcripts in the developing central and peripheral nervous systems. Translation of sna RNA is apparently delayed as the sna protein is not detected before the onset of gastrulation. Its regional distribution generally correlates with that of sna transcripts. The complex pattern of sna expression strongly suggests that the function of the gene is not restricted to mesoderm formation.