bluuebunny/crossref_metadata_2025_split huggingface dataset:
GATA-4 is a novel transcription factor expressed in endocardium of the developing heart
Authors: Kelley Clair , Blumberg Heather , Zon Leonard I. , Evans Todd 
Published: April 2021

ABSTRACT
We have isolated and characterized Xenopus cDNA clones for a new transcription factor that represents an early marker for the developing heart. The cDNAs encode a protein that we have designated GATA-4; it contains the highly conserved DNA-binding domain that characterizes this family of cell-type restricted transcriptional activators. Whole-embryo in situ analysis of Xenopus embryos demonstrates that the GATA-4 gene is transcribed in presumptive cardiac ventral mesoderm at the time that bilateral progenitors fuse and form the cardiac tube. GATA-4 is therefore the earliest molecular marker of cardiogenesis yet characterized. By stage 30, the GATA-4 mRNA is expressed in the developing atria and ventricles; at stage 38, cross-sections reveal that the gene is active in the endocardial layer, but not in myocardium. By stage 40, GATA-4 message is detected in the great vessels. In the adult frog, the GATA-4 gene is highly transcribed in heart and gut; lower levels of message are detected in various endoderm-derived tissues and gonads. Expression in the stomach is largely confined to the epithelium. The GATA-4 gene is first activated at stage 11; mRNA is initially present throughout the marginal zone of explants and later partially localized to the ventral marginal zone. GATA-4 mRNA is also detected at high levels in cultured endodermal explants derived from the vegetal region of early embryos. In mesoderm induction experiments, GATA-4 transcription is not induced in animal caps treated with activin or bFGF. The GATA-4 gene may provide a new early marker for studying the inductive processes that lead to the formation of the cardiovascular system and for the specification of the endocardial lineage.
Precursor cells of mouse endocrine pancreas coexpress insulin, glucagon and the neuronal proteins tyrosine hydroxylase and neuropeptide Y, but not pancreatic polypeptide
Authors: Teitelman G. , Alpert S. , Polak J. M. , Martinez A. , Hanahan D. 
Published: April 2021

ABSTRACT
The early progenitor cells to the pancreatic islets in the mouse have been characterized so as to re-examine their possible lineage relationships to the four islet cell types found in mature islets. Insulin and glucagon were both first expressed at embryonic day 9.5, and many cells co-expressed these two markers, as shown by light and electron microscopic analysis using double-label immunohistochemistry. Incubation of embryonic pancreas with 1% glutaraldehyde, a fixative commonly used by electron microscopists, abolished this reactivity, thereby explaining reported difficulties in detecting these precursor cells. Using antisera specific for neu-ropeptide Y (NPY) a peptide with considerable homology to pancreatic polypeptide (PP), we show that NPY first appears with insulin and glucagon immunore-activity at E9.5, and is co-expressed with glucagon in a majority of adult cells. As we have previously reported, PP itself is first detectable immunocytochemically at postnatal day 1 with PP-specific antibodies. However, antibodies raised against bovine PP are shown by dot blotting to recognize NPY with comparable avidity, indi-cating that a recent report of islet progenitor cells containing PP at E9.5 (Herrera, P. L., Huarte, J., Sanvito, F., Meda, P., Orci, L. and Vassalli, J. D. (1991) Devel-opment 113, 1257-1265), actually represents cross-reactivity to NPY. The data support a model in which early precursor cells to the endocrine pancreas co-activate and co-express a set of islet cell hormone and neural genes, whose expression is both selectively increased and extinguished as development proceeds, concomitant with a restriction to the patterns of expression characteristic of mature islet cell types.
A fourth human MEF2 transcription factor, hMEF2D, is an early marker of the myogenic lineage
Authors: Breitbart Roger E. , Liang Chang-seng , Smoot Leslie B. , Laheru Dan A. , Mahdavi Vijak , Nadal-Ginard Bernardo 
Published: April 2021

ABSTRACT
The transition from multipotent mesodermal precursor to committed myoblast and its differentiation into a mature myocyte involve molecular events that enable the cell to activate muscle-specific genes. Among the participants in this process is the myocyte-specific enhancer factor 2 (MEF2) family of tissue-restricted transcription factors. These factors, which share a highly conserved DNA-binding domain including a MADS box, are essential for the expression of multiple muscle genes with cognate target MEF2 sites in cis. We report here a new human MEF2 factor, hMEF2D, which is unique among the members of this family in that it is present not only in myotubes but also in undifferentiated myoblasts, even before the appearance of myogenin. hMEF2D comprises several alternatively spliced products of a single gene, one of which is the human homolog of the Xenopus SRF-related factor SL-1. Like its relatives, cloned hMEF2D is capable of activating transcription via sequence-specific binding to the MEF2 site, recapitulating endogenous tissue-specific MEF2 activity. Indeed, while MEF2D mRNAs are ubiquitous, the protein is highly restricted to those cell types that contain this activity, implicating posttranscriptional mechanisms in the regulation of MEF2D expression. Alternative splicing may be important in this process: two alternative MEF2D domains, at least one of which is specifically included during myogenic differentiation, also correlate precisely with endogenous MEF2 activity. These findings provide compelling evidence that MEF2D is an integral link in the regulatory network for muscle gene expression. Its presence in undifferentiated myoblasts further suggests that it may be a mediator of commitment in the myogenic lineage.
Basic Fibroblast Growth Factor (bFGF) acts intracellularly to cause the transdifferentiation of avian neural crest-derived Schwann cell precursors into melanocytes
Authors: Sherman Larry , Stocker Kate M. , Morrison Richard , Ciment Gary 
Published: April 2021

ABSTRACT
We previously found that cultured neural crest-derived cells from embryonic quail peripheral nerves, which consist mostly of Schwann cell precursors, gave rise to melanocytes following treatment with basic fibroblast growth factor (bFGF) or 12-O-tetradecanoyl phorbol-13-acetate (TPA). Here, we show that antisense deoxyoligonucleotides targeted against two regions of the bFGF mRNA transcript blocked this TPA-induced transdifferentiation of Schwann cell precursors. Neither sense nor scrambled antisense control oligonucleotides had any effect in this regard. TPA increased bFGF protein expression in cell lysates but not in conditioned media from these cultures, and this expression was localized to the nucleus and cytoplasm. Furthermore, bFGF-neutralizing antibodies and inositolhexakisphosphate (InsP6) both inhibited pigmentation caused by exogenous bFGF, but had no affect on TPA-induced melanogenesis, suggesting that bFGF is not released by these cells. These data indicate that bFGF is necessary for the TPA-induced transdifferentiation of Schwann cell precursors into melanocytes and that bFGF acts via an intracrine mechanism.
Fertilisation and thimerosal stimulate similar calcium spiking patterns in mouse oocytes but by separate mechanisms
Authors: Cheek Timothy R. , McGuinness Orla M. , Vincent Caroline , Moreton Roger B. , Berridge Michael J. , Johnson Martin H. 
Published: April 2021

ABSTRACT
Exposure of freshly ovulated mouse oocytes to a fertilising spermatozoon, thimerosal, Sr2+ or acetylcholine induced similar Ca2+ spiking responses. We propose that each of the four agents reduces the threshold for Ca2+ release from internal stores, but by different mechanisms.
All agents except thimerosal stimulated oocyte activation, but thimerosal caused dissassembly of the meiotic spindle and thus prevented progress into interphase. Dithiothreitol (DTT) completely blocked and reversed the spiking responses induced by thimerosal, but facilitated and accelerated those induced by spermatozoa, Sr2+ and acetylcholine. The stimulatory effect of DTT was not simply a consequence of progress into interphase, but was attributable, at least in part, to an enhancement of divalent cation entry, as measured by Mn2+ quench analysis of fura-2 in both fertilised and unfertilised oocytes. Possible mechanisms by which DTT might achieve its effects are discussed.
Even-numbered rhombomeres control the apoptotic elimination of neural crest cells from odd-numbered rhombomeres in the chick hindbrain
Authors: Graham Anthony , Heyman Isobel , Lumsden Andrew 
Published: April 2021

ABSTRACT
Neural crest cells originate at three discontinuous levels along the rostrocaudal axis of the chick rhomben-cephalon, centred on rhombomeres 1 and 2, 4 and 6, respectively. These are separated by the odd-numbered rhombomeres r3 and r5 which are depleted of migratory neural crest cells. Here we show elevated levels of apoptosis in the dorsal midline of r3 and r5, immediately following the formation of these rhombomeres at the developmental stage (10-12) when neural crest cells would be expected to emerge at these neuraxial levels. These regions are also marked by their expression of members of the msx family of homeobox genes with msx-2 expression preceding apoptosis in a precisely co-localised pattern. In vitro and in ovo experiments have revealed that r3 and r5 are depleted of neural crest cells by an interaction within the neural epithelium: if isolated or distanced from their normal juxtaposition with even-numbered rhombomeres, both r3 and r5 produce migrating neural crest cells. When r3 or r5 are unconstrained in this way, allowing production of crest, msx-2 expression is concomitantly down regulated. This suggests a correlation between msx-2 and the program-ming of apoptosis in this system. The hindbrain neural crest is thus produced in discrete streams by mechanisms intrinsic to the neural epithelium. The crest cells that enter the underlying branchial region are organised into streams before they encounter the mesodermal environ-ment lateral to the neural tube. This contrasts sharply with the situation in the trunk where neural crest pro-duction is uninterrupted along the neuraxis and the segmental accumulation of neurogenic crest cells is sub-sequently founded on an alternation of permissive and non-permissive qualities of the local mesodermal envi-ronment.
Cardiovascular development in the zebrafish I. Myocardial fate map and heart tube formation
Authors: Stainier Didier Y. R. , Lee Robert K. , Fishman Mark C. 
Published: April 2021

ABSTRACT
We have analyzed the origin of cardiac progenitors in the zebrafish embryo by injection of single blastomeres with a lineage tracer dye, and examined the formation of the zebrafish heart tube by serial sectioning of immunostained embryos. At the 512-cell stage (early blastula), most cardiac progenitors lie in a marginal zone that extends from 90° longitude (midway between the future dorsal and ventral axis) through 180° longitude (the future ventral axis) to 270° longitude. By focusing on myocardial progenitors located at 90° (and 270°) longitude, we found that a single cell injected in the early blastula can contribute progeny to both the atrium and ventricle. A cell injected in the midblastula contributes progeny to either the atrium or ventricle, but not both. This analysis suggests that, at least for these myocardial progenitors, the atrial and ventricular lineages separate in the midblastula.
Precardiac cells involute early during gastrulation and turn towards the animal pole with other early involuting cells. These cardiogenic cells reach the embryonic axis around the 8-somite stage, and there they coalesce to form a pair of myocardial tubular primordia on either side of the midline. By the 21-somite stage, the tropomyosin-immunoreactive myocardial tubes have moved closer to each other, and a distinct group of cells, the endocardial progenitor cells, sits medially between them. The myocardial tubes then fuse to enclose the endocardial cells and form the definitive heart tube. By 22 hours postfertilization (26-somite stage), the heart tube is clearly beating. The regionalization of cardiac myosin heavy chain expression distinguishes the cardiac chambers at this stage, although they are not morpho-logically delineated until 36 hours.
This work shows that cardiogenic regions can be identified in the early blastula, and that chamber restriction seems to arise in the midblastula. Additionally, it provides the basis for embryological perturbation at the single cell level, as well as for the genetic analysis of heart tube formation in the zebrafish.
The zebrafish midblastula transition
Authors: Kane Donald A. , Kimmel Charles B. 
Published: April 2021

ABSTRACT
The zebrafish midblastula transition (MBT) begins at cycle 10. It is characterized by cell cycle lengthening, loss of cell synchrony, activation of transcription and appearance of cell motility. Superceding a 15 minute oscillator that controls the first nine cycles, the nucleocytoplasmic ratio appears to govern the MBT. This timing mechanism operates cell autonomously: clones of labeled cells initiate cell cycle lengthening independently of neighbors but dependent on immediate lineal ancestors. Unequal divisions, when they occur, produce asymmetric cell cycle lengthening based on the volume of each daughter. During the several cycles after the MBT begins, cycle length is correlated with the reciprocal of the blastomere volume, suggesting a continuation of cell cycle regulation by the nucleocytoplasmic ratio during an interval that we term the ‘MBT period’.
Expression and binding characteristics of the BDNF receptor chick trkB
Authors: Dechant Georg , Biffo Stefano , Okazawa Hitoshi , Kolbeck Roland , Pottgiesser Jutta , Barde Yves-Alain 
Published: April 2021

ABSTRACT
Previous studies using transfected cells have indicated that the mammalian receptor tyrosine kinase trkB binds the neurotrophins brain-derived neurotrophic factor, neurotrophin-3 and neurotrophin-4. However, most studies demonstrating that these neurotrophins prevent the death of embryonic neurons and have specific neuronal receptors have been performed with chick neurons. In order to explore the possibility that trkB is the molecular entity representing the high-affinity receptor for brain-derived neurotrophic factor on embryonic chick neurons, we cloned and expressed a chick trkB cDNA. In situ hybridisation results indicate that the distribution of trkB mRNA in the peripheral nervous system of the developing chick embryo correlates well with the structures known to respond to brain-derived neurotrophic factor. Binding studies performed with a cell line stably transfected with the ctrkB cDNA indicate a dissociation constant for brain-derived neurotrophic factor of 9.9 10 10 M, which is distinctly higher than that found on primary chick sensory neurons (1.5×10−11 M). When binding of brain-derived neurotrophic factor was determined in the presence of other neurotrophins, neurotrophin-3 was found efficiently to prevent the binding of brain-derived neurotrophic factor to both the ctrkB cell line and embryonic sensory neurons. In vitro, neurotrophin-3 at high concentrations completely blocked the survival normally seen with brain-derived neurotrophic factor. Thus, unlike previous cases of receptor occupancy by heterologous neurotrophins (which resulted in agonistic effects), the interaction between the brain-derived neurotrophic factor receptor and neurotrophin-3 on sensory neurons is antagonistic.
Spatial and temporal changes in the expression of fibroglycan (syndecan-2) during mouse embryonic development
Authors: David Guido , Bai Xiao Mei , Schueren Bernadette Van der , Marynen Peter , Cassiman Jean-Jacques , Berghe Herman Van den 
Published: April 2021

ABSTRACT
Fibroglycan (syndecan-2) is a member of a family of cell surface heparan sulfate proteoglycans that interact with adhesion molecules, growth factors and a variety of other effector systems that support the shaping, maintenance and repair of an organism. To investigate this apparent redundancy of proteoglycans at the cell surface, we have studied the expression of fibroglycan in the mouse embryo and compared this expression with that of syndecan-1. The characterisation of mouse embryo cDNA clones that crosshybridized to human fibroglycan-cDNA predicted that murine and human fibroglycan were highly similar in structure. Consistently, the analysis of transfectant cells, murine cell lines and embryo extracts indicated that the murine proteoglycan reacted specifically with monoclonal antibody 10H4 developed against the human protein. Fibroglycan, as detected by monoclonal antibody 10H4 in sections of embryonic tissues, occurred exclusively on mesenchymal cells that represented the putative precursors of the hard and connective tissue cells. No fibroglycan was detected in epithelia or in muscle cells. Areas where fibroglycan was particularly abundant were sites of high morphogenetic activity where intense cell-cell and cell-matrix interactions are known to occur (e.g. the epithelial-mesenchymal interfaces, the prechondrogenic and preosteogenic mesenchymal condensations). The expression of fibroglycan was weak in the early embryo, culminated during the morphogenetic phase and at the moment of cell lineage differentiation, and persisted in the perichondrium, periosteum and connective tissue cells. Syndecan-1, in contrast, was primarily detected in epithelia, and transiently in some mesenchymal cells, with mesenchymal localisations that did not or only partially overlap with those of fibroglycan. In situ hybridization analyses confirmed these expression patterns at the transcriptional level, identifying mesenchymal cells as the major source of fibroglycan production. These data indicate that the expression of fibroglycan occurs along unique and developmentally regulated patterns, and suggest that fibroglycan and syndecan-1 may have distinctive functions during tissue morphogenesis and differentiation.
Defects in mesoderm, neural tube and vascular development in mouse embryos lacking fibronectin
Authors: George Elizabeth L. , Georges-Labouesse Elisabeth N. , Patel-King Ramila S. , Rayburn Helen , Hynes Richard O. 
Published: April 2021

ABSTRACT
To examine the role of fibronectin in vivo, we have generated mice in which the fibronectin gene is inactivated. Heterozygotes have one half normal levels of plasma fibronectin, yet appear normal. When homozygous, the mutant allele causes early embryonic lethality, proving that fibronectin is required for embryogenesis. However, homozygous mutant embryos implant and initiate gastrulation normally including extensive mesodermal movement. Neural folds also form but the mutant embryos subsequently display shortened anterior-posterior axes, deformed neural tubes and severe defects in mesodermally derived tissues. Notochord and somites are absent; the heart and embryonic vessels are variable and deformed, and the yolk sac, extraembryonic vasculature and amnion are also defective. These abnormalities can be interpreted as arising from fundamental deficits in mesodermal migration, adhesion, proliferation or differentiation as a result of the absence of fibronectin. The nature of these embryonic defects leads to reevaluation of suggested roles for fibronectin during early development based on results obtained in vitro and in embryos of other species.
Normal development of mice carrying a null mutation in the gene encoding the L14 S-type lectin
Authors: Poirier Françoise , Robertson Elizabeth J. 
Published: April 2021

ABSTRACT
The L14 lectin is a 14 103Mr carbohydrate binding protein belonging to the family of S-type lectins. The pattern of expression of this protein during mouse embryogenesis suggests that it may have multiple roles during preand post-implantation development. Using the technique of homologous recombination in embryonic stem cells, we have introduced a null mutation in the gene encoding the L14 lectin and generated a strain of mice carrying the mutant allele. We report here that homozygous mutant animals that lack the L14 lectin develop normally and are viable and fertile. The absence of any major phenotypic abnormalities in these mutant animals suggests that other protein(s) potentially compensate for the absence of the L14 lectin. Here we show that a related protein termed L30, a lectin that has carbohydrate binding specificity similar to that of L14, is present in the same embryonic cell populations as L14 at the time of implantation, suggesting that the two S-type lectins may be capable of functional substitution at this early stage of embryogenesis.
Taurine promotes the differentiation of a vertebrate retinal cell type in vitro
Authors: Altshuler David , Lo Turco Joseph J. , Rush John , Cepko Constance 
Published: April 2021

ABSTRACT
The retina offers a model system for investigating the mechanisms that control cell type determination and differentiation in the vertebrate central nervous system. Previously, rod photoreceptor development in vitro was found to require a diffusible activity released by retinal cells (D. Altshuler and C. Cepko, Development 114, 947957, 1992). In this report, we show that retinal-cell-conditioned medium and extracts contain two separable activities that influence rod development: a &gt;10 kDa inhibitory activity, and a stimulatory activity that is &lt;1 kDa and heat stable. Taurine was found to be a component of the &lt;1 kDa fraction and to stimulate rod development when added to retinal cultures. Taurine was not the only rod-promoting factor in these retinal preparations, however, as conditioned medium and extracts stimulated a higher level of rod development than did taurine alone. Taurine uptake into cells could be blocked without inhibiting taurine’s ability to stimulate rod development, arguing against an osmoregulatory or nutritive mechanism of action. Finally, a competitive antagonist of taurine’s bioactivity was identified and shown partially to inhibit rod development in retinal explants, suggesting that taurine may normally act to stimulate rod development in the retina. These results provide evidence for three activities, one of which is taurine, that are candidate regulators of rod photoreceptor development in vivo.
Nuclear localization of the protein encoded by the Wilms’ tumor gene WT1 in embryonic and adult tissues
Authors: Mundlos Stefan , Pelletier Jerry , Darveau André , Bachmann Michael , Winterpacht Andreas , Zabel Bernhard 
Published: April 2021

ABSTRACT
The human Wilms’ tumor gene WT1 encodes a putative transcription factor implicated in tumorigenesis and in specifying normal urogenital development. We have studied the distribution of WT1 protein and mRNA using immunohistochemistry and in situ hybridization.
Monoclonal antibodies were raised against a peptide specific to the first alternative splice site of WT1. Two antibodies specifically reacted on Western blot to this WT1 isoform. Immunofluorescence localized WT1 protein to podocytes during mesonephric and metanephric development. In situ hybridization revealed a similar pattern of expression except that WT1 mRNA was also present in metanephric blastema and renal vesicles. Messenger RNA expression was most pronounced in the kidneys during early fetal development and declined thereafter. In contrast, WT1 protein was readily detectable in glomerular podocytes throughout adulthood. WT1 protein in Wilms’ tumor was present in blastema and glomeruloid structures.
Expression in the female gonad was linked to the different stages of granulosa cell development. In the male gonad, expression was restricted to Sertoli cells and their precursors, the embryonic tunica albuginea and the rete testis.
The intracellular distribution of the WT1 protein was investigated by confocal laser microscopy and was demonstrated to be exclusively nuclear.
The nuclear distribution and the selective pattern of expression support the proposed role of WT1 as a transcription factor active during urogenital development. The persistence of WT1 expression in the adult kidney suggests a role in homeostasis of the podocyte.