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PubMed1986

Ito's hypomelanosis (incontinentia pigmenti achromians). A review of four cases.

In 1952 Ito described the occurrence of a bilateral systematized depigmented nevus in a 22-year-old Japanese woman. He used the term incontinentia pigmenti achromians. The condition has been described under various designations, such as for instance Ito's hypomelanosis. Till now 71 patients with this syndrome are described. We will report 4 cases, 2 boys and 2 girls, 2 Caucasian, 1 Indonesian and 1 Caribean child. The cutaneous signs in these 4 patients fit in with the syndrome of Ito's hypomelanosis. Of these 4 children 3 are mentally retarded, 2 have epilepsy. Congenital malformations are seen in 3 children. Electronmicroscopy of skin biopsies of the hypomelanotic nevus and of the normal skin were performed. In the biopsy of the normal skin of one patient interruption of the basement membrane is seen. Anomalies of the central nervous system as seen in our patients occur in about 40% of the cases. Abnormalities of skin derivatives next to other ectodermal anomalies are described. Affection of other germ layers also occur to a varying degree. In our 4 patients some of these abnormalities exist also. These 4 cases are presented to underline the fact that this syndrome seems not to be as extremely rare as is proposed.

Abnormalities, Multiple↗

PubMed1984

The synthesis of 5S RNA and transfer RNA in sea urchin embryos animalized by Evans blue.

Animalization (ectodermalization) of embryos of Arbacia punctulata was induced by continuous treatment from the 2-cell stage with Evans Blue. Quantitative measurements of rates of accumulation of 5S RNA and tRNA in mesenchyme blastulae, in plutei and in corresponding animalized embryos showed that these rates per embryo and per cell were similar in control and in animalized embryos. The alterations in normal cell interactions and germ layer formation induced by Evans Blue did not affect the synthesis of 5S RNA and tRNA in sea urchin embryos.

Animals↗

PubMed1993

The Drosophila melanogaster developmental gene g1 encodes a variant zinc-finger-motif protein.

In Drosophila melanogaster, the mechanisms involved in the pattern formation of complex internal organs are still largely unknown. However, the identity of the molecular determinants that control the development of these specific tissues is emerging from the combined use of genetic and molecular approaches. We have cloned a gene that is expressed in the mesoderm, one of the fundamental embryonic germ layers which gives rise to internal structures, such as the musculature. Here, we describe the molecular characterization of this gene, designated as g1. The nucleotide (nt) sequence of its cDNA shows an open reading frame of 852 nt, which encodes a 32-kDa protein with two putative zinc fingers, and a serine/glutamine/proline-rich region. These features indicate a functional role for g1, which remains to be elucidated, in regulating gene expression during mesoderm formation.

Amino Acid Sequence↗

PubMed1990

Spatio-temporal distribution of the adherens junction-associated molecules vinculin and talin in the early avian embryo.

To gain an insight into the possible involvement of the cytoskeletal components and cellular junctions in morphogenetic processes during development, we have studied the spatio-temporal distribution of two major adherens-junction-associated molecules, vinculin and talin, during avian embryogenesis, using immunofluorescence microscopy and immunoblotting. Both molecules were detected at very early stages during morphogenesis and were found in a wide variety of tissues deriving from the three primary germ layers. A number of tissues, including smooth and striated muscles, endothelia, and some hemopoietic precursors, expressed vinculin and talin at especially high levels either transiently or permanently. Conversely, only a few cell types, e.g., circulating erythrocytes and neurones in the central nervous system lacked or expressed them at very low levels. In addition, expression of vinculin and talin was in some cases modulated in connection with morphological rearrangements of tissues. In particular, they were transiently enhanced in restricted areas of the ectoderm and endoderm undergoing extensive foldings. However, other morphogenetic events such as local disruptions of epithelia were not accompanied by extensive modifications in their expression. Finally, it appeared that, in most cases, vinculin and talin overlapped in their distribution, and the level of their expression was regulated coincidently with the notable exceptions of the primordium of the central nervous system, the nephron, and the liver where each molecule followed independent regulatory patterns. It appears from this study that the spatio-temporal distribution of vinculin and talin correlates frequently with that of the adhesion molecules A-CAM (or N-cadherin), L-CAM, and of integrin receptors. Thus, vinculin and talin, in association with the membrane components of adherens junctions, may actively participate both in the control of cellular interactions during early embryonic development and in cell differentiation.

Animals↗

PubMed1992

Retinoic acid prevents accumulation of a mesoderm-specific mRNA in the amphibian embryo.

Our greatest progress in understanding mechanisms of germ layer specification in vertebrates has come from studies of mesoderm induction in amphibian embryos. A key early observation was that transcriptional activation of the muscle actin gene and accumulation of its mRNA can be used as molecular markers of mesoderm induction. Recent studies of putative amphibian morphogens have shown that exposure to all-trans retinoic acid (RA) can perturb both neural and mesodermal patterning in the Xenopus embryo, and that RA can, under certain conditions, induce the synthesis of mRNA for a gene whose product is able to influence the patterning of the embryo. To characterize further the mechanism of mesoderm induction, the effect of RA exposure on the transcriptional properties of the muscle actin gene has been investigated. Interestingly, prior exposure of embryos to RA appears to inhibit accumulation, rather than initial activation, of the mesoderm-specific muscle actin transcript. This effect is concentration-dependent and is not observed with the ubiquitously expressed genes for cytoskeletal actin and translation elongation factor 1 alpha, nor with a neural-specific beta-tubulin gene. Thus, the molecular mechanism of mesoderm induction may include, in addition to transcriptional activation of specific genes, processes involved in maintenance of mRNA accumulation for those gene products.

Actins↗

PubMed1992

Aggregation and cell cycle dependent retinoic acid receptor mRNA expression in P19 embryonal carcinoma cells.

Differentiation of P19 EC cells along different pathways into derivatives resembling cells of the three embryonic germ layers is accompanied by characteristic differences in modulation of expression of each of the three retinoic acid receptor genes, RAR alpha, -beta and -gamma. Differentiation induced by addition of RA to P19 EC cells cultured in monolayer is accompanied by a rapid increase in expression of both RAR alpha and -beta. Induction of RAR beta occurs in a characteristic biphasic manner, suggesting that multiple factors and/or different mechanisms are involved in controlling its expression. RAR beta mRNA is induced to a far higher level during early aggregation in the presence of RA than during early differentiation in monolayer, suggesting that the direction of differentiation depends on the number and/or ratio of alpha and beta type of RA receptors. Aggregation of P19 EC cells in the presence of RA, but not DMSO, is accompanied by repression of RAR gamma, suggesting that the expression of RAR beta and RAR gamma during neuroectodermal differentiation is mutually exclusive. The effects of RA on RAR expression are significantly greater in G1 than in S-phase of the cell cycle. These results extend previous observations that commitment to differentiation is cell cycle dependent and indicates that critical target gene regulation in response to RA has to take place in G1 for differentiation to occur.

Carrier Proteins↗

PubMed1993

Murine Cdx-4 bears striking similarities to the Drosophila caudal gene in its homeodomain sequence and early expression pattern.

A third member of the murine caudal-like gene family, Cdx-4, has been isolated. In situ hybridization and immunohistochemistry have been used to study the localization of mRNA and protein during murine embryogenesis. Cdx-4 is expressed transiently from 7.0 d.p.c. (days post coitum) until 10 d.p.c., starting at the beginning of gastrulation (7.0-7.5 d.p.c.) in the allantois and posterior tip of the primitive streak. At the mid-streak stage, Cdx-4 expression moves rostrally, and protein and mRNA are detected in all cells over the posterior half of the primitive streak. As development proceeds, Cdx-4 gene products continue to be restricted to the posterior of the embryo, including the remnants of the primitive streak. Cdx-4 is expressed in neurectoderm, presomitic and lateral plate mesoderm, and hindgut endoderm, but the anterior boundary in the paraxial mesoderm is staggered with respect to the other germ layers. At all stages analyzed, Cdx-4 exhibits a graded expression pattern with a posterior maximum, a distribution highly reminiscent of the Drosophila caudal gene. These data add to the recently described distributions of other vertebrate caudal-like genes, and further support the idea that members of this homeobox gene subfamily have regulatory roles in the specification of anteroposterior axial polarity in early embryos.

Amino Acid Sequence↗

PubMed1993

The pattern of retinoic acid receptor gamma (RAR gamma) expression in normal development of Xenopus laevis and after manipulation of the main body axis.

Nuclear receptors are a prerequisite for the transduction of retinoid signals in vertebrate embryogenesis. We have raised an antiserum against the X. laevis retinoic acid receptor gamma (xRAR gamma), which specifically detects a polypeptide with an M(r) of 54 x 10(3) in embryos at gastrula and neurula stages. The antiserum reveals xRAR gamma as a nuclear protein in the head endomesoderm, the neuroectoderm of the hindbrain region, and the entire posterior region during neurula stages. Explanted animal caps express low amounts of xRAR gamma in the absence of mesoderm induction. Treatment of animal caps with activin leads to an increase of the amount and to a regionalization of the xRAR gamma expression. In exogastrulae and in Keller sandwiches, the pattern of expression in the endomesoderm is as expected by analogy to the normal embryo and therefore independent of the vertical contact between the germ layers. A planar signal coming from the dorsal mesoderm is sufficient to impose region-specific expression of xRAR gamma in the neuroectoderm of Keller sandwiches.

Activins↗

PubMed1994

Cloning and developmental expression of LFB3/HNF1 beta transcription factor in Xenopus laevis.

We have cloned the Xenopus laevis homologue of the LFB3/HNF1 beta transcription factor. RNase protection and in situ hybridisation experiments show that XLFB3 transcription starts in the gastrulating endoderm at stage 10.5 (mid-gastrula). At later stages, XLFB3 transcripts within the endoderm are restricted to mid- and hindgut and to their derivative organs and tissues. XLFB3 is also expressed in the neuroectoderm and in the pronephros anlage. XLFB3 is not expressed in the rostral part of all three germ layers, with coincident anterior borders that are shifted anteriorly by treatment of developing embryos with retinoic acid. XLFB3 is a useful marker of early endoderm differentiation and its expression pattern along the antero-posterior axis, as well as the response to retinoic acid treatment, suggests a role in early morphogenesis.

Amino Acid Sequence↗

PubMed1995

Homeobox genes in vertebrate gastrulation.

The formation and anteroposterior patterning of the three definitive germ layers, ectoderm, or epiblast, is the common theme of vertebrate gastrulation. What changes from system to system is the geometry of these events and the nature of the non-epiblast transient structures implicated. A number of molecular markers, including a few homeobox genes and in particular goosecoid and Otx2, are now available that will hopefully allow us to explore the underlying molecular mechanisms and to establish biologically relevant homologies between the various systems.

Animals↗

PubMed2006

Adult pancreatic stem/progenitor cells spontaneously differentiate in vitro into multiple cell lineages and form teratoma-like structures.

Cells isolated from pancreas have a remarkable potential for self-renewal and multilineage differentiation. We here present a comprehensive characterisation of stem/progenitor cells derived from exocrine parts of the adult rat pancreas. Using purified cells from either single colonies or even single-cell clones, we specifically demonstrate: (i) the cells contain the typical stem/progenitor cell markers alkaline phophatase, SSEA-1, Oct-4, CD9, Nestin, Pax6, CD44, a-Fetoprotein and Brachyury, demonstrated by immunocytochemistry and RT-PCR; (ii) the cells have the potential to differentiate into lineages of all three germ layers in vitro; (iii) a clonal analysis revealed that even cell lines derived from a single cell have stem/progenitor cell properties such as self-renewal and spontaneous differentiation into various cell lineages; (iv) the cells have the propensity to form three-dimensional, teratoma-like structures in vitro, which contain cells of different lineages; and (v) external stimuli can activate the generation of certain cell types. For instance, cells treated with retinoic acid show an increased expression of alpha-smooth muscle actin. These results suggest that exocrine glands, such as pancreas may be a potential source of adult stem/progenitor cells, suitable for cell therapy of degenerative diseases.

Actins↗

PubMed2006

Ca2+ signaling and early embryonic patterning during the blastula and gastrula periods of zebrafish and Xenopus development.

It has been proposed that Ca(2+) signaling, in the form of pulses, waves and steady gradients, may play a crucial role in key pattern forming events during early vertebrate development [L.F. Jaffe, Organization of early development by calcium patterns, BioEssays 21 (1999) 657-667; M.J. Berridge, P. Lipp, M.D. Bootman, The versatility and universality of calcium signaling, Nat. Rev. Mol. Cell Biol. 1 (2000) 11-21; S.E. Webb, A.L. Miller, Calcium signalling during embryonic development, Nat. Rev. Mol. Cell Biol. 4 (2003) 539-551]. With reference to the embryos of zebrafish (Danio rerio) and the frog, Xenopus laevis, we review the Ca(2+) signals reported during the Blastula and Gastrula Periods. This developmental window encompasses the major pattern forming events of epiboly, involution, and convergent extension, which result in the establishment of the basic germ layers and body axes [C.B. Kimmel, W.W. Ballard, S.R. Kimmel, B. Ullmann, T.F. Schilling, Stages of embryonic development of the zebrafish, Dev. Dyn. 203 (1995) 253-310]. Data will be presented to support the suggestion that propagating waves (both long and short range) of Ca(2+) release, followed by sequestration, may play a crucial role in: (1) Coordinating cell movements during these pattern forming events and (2) Contributing to the establishment of the basic embryonic axes, as well as (3) Helping to define the morphological boundaries of specific tissue domains and embryonic structures, including future organ anlagen [E. Gilland, A.L. Miller, E. Karplus, R. Baker, S.E. Webb, Imaging of multicellular large-scale rhythmic calcium waves during zebrafish gastrulation, Proc. Natl. Acad. Sci. USA 96 (1999) 157-161; J.B. Wallingford, A.J. Ewald, R.M. Harland, S.E. Fraser, Calcium signaling during convergent extension in Xenopus, Curr. Biol. 11 (2001) 652-661]. The various potential targets of these Ca(2+) transients will also be discussed, as well as how they might integrate with other known pattern forming pathways known to modulate early developmental events (such as the Wnt/Ca(2+)pathway; [T.A. Westfall, B. Hjertos, D.C. Slusarski, Requirement for intracellular calcium modulation in zebrafish dorsal-ventral patterning, Dev. Biol. 259 (2003) 380-391]).

Animals↗

PubMed2005

The in vitro differentiation of rat neural stem cells into an insulin-expressing phenotype.

Mature beta-cells and nerve cells share many functional similarities despite originating from different embryonic germ layers. The aim of this study was to investigate the potential of neural stem cells (NSCs), isolated from foetal rat brain, as a starting material from which to generate functionally responsive, insulin-containing cells. Our results demonstrated that NSCs can be significantly expanded in vitro and can be induced to express increased preproinsulin mRNA levels. In addition, these NSC-derived cells expressed transcriptional and functional elements associated with a mature beta-cell phenotype. The differentiated cells showed functional responses typical of pancreatic beta-cells, including glucose-dependent increases in metabolism and rapid elevations in intracellular Ca(2+) in response to the sulphonylurea tolbutamide or to increased glucose concentration. These results suggest that NSCs may have potential as a starting material from which to generate beta-cell surrogates for the treatment of patients with Type 1 diabetes mellitus.

Animals↗

PubMed2006

A novel chemical-defined medium with bFGF and N2B27 supplements supports undifferentiated growth in human embryonic stem cells.

Traditionally, undifferentiated human embryonic stem cells (hESCs) are maintained on mouse embryonic fibroblast (MEF) cells or on matrigel with an MEF-conditioned medium (CM), which hampers the clinical applications of hESCs due to the contamination by animal pathogens. Here we report a novel chemical-defined medium using DMEM/F12 supplemented with N2, B27, and basic fibroblast growth factor (bFGF) [termed NBF]. This medium can support prolonged self-renewal of hESCs. hESCs cultured in NBF maintain an undifferentiated state and normal karyotype, are able to form embryoid bodies in vitro, and differentiate into three germ layers and extraembryonic cells. Furthermore, we find that hESCs cultured in NBF possess a low apoptosis rate and a high proliferation rate compared with those cultured in MEF-CM. Our findings provide a novel, simplified chemical-defined culture medium suitable for further therapeutic applications and developmental studies of hESCs.

Apoptosis↗

PubMed2004

Stem cell: balancing aging and cancer.

Stem cells are defined by their self-renewing capacity and the ability to differentiate into one or more cell types. Stem cells can be divided, depending on their origin, into embryonic or adult. Embryonic stem cells derive from early stage embryos and can give rise to cells from all three germ layers. Adult stem cells, first identified in hematopoietic tissue, reside in a variety of adult tissues. Under normal physiologic conditions, adult stem cells are capable of differentiating into the limited cell types that comprise the particular tissue or organ. Adult stem cells are responsible for tissue renewal and exhaustion of their replicative capacity may contribute to tissue aging. Loss of unlimited proliferative capacity in some of the adult stem cells and/or their progenitors may have involved the evolutionary trade-off: senescence prevents cancer but may promote aging. Embryonic stem cells exhibit unlimited self-renewal capacity due to the expression of telomerase. Although they possess some cancer cell characteristics, embryonic stem cells exhibit a remarkable resistance to genomic instability and malignant transformation. Understanding the tumor suppressive mechanisms employed by embryonic stem cells may contribute to the development of novel cancer treatments and safe cell-based therapies for age-related diseases.

Adult↗

PubMed2006

Low oxygen tension inhibits osteogenic differentiation and enhances stemness of human MIAMI cells.

We recently reported the isolation of a unique subpopulation of human stromal cells from bone marrow (BM) termed marrow-isolated adult multilineage inducible (MIAMI) cells, capable of differentiating in vitro into mature-like cells from all three germ layers. The oxygen tension (pO2) in BM ranges from 1 to 7%, which prompted us to examine the role of pO2 in regulating the capacity of MIAMI cells both to self-renew and maintain their pluripotentiality (stemness) or to progress toward osteoblastic differentiation. MIAMI cells were grown under low-pO2 conditions (1, 3, 5, and 10% oxygen) or air (21% oxygen). The proliferation rate of cells exposed to 3% oxygen (3 days) increased, resulting in cell numbers more than threefold higher than those of cells exposed to air (at 7 days). In cells grown under osteoblastic differentiation conditions, the expression of the osteoblastic markers osteocalcin, bone sialoprotein, osterix, and Runx2 and alkaline phosphatase activity was upregulated when incubated in air; however, it was blocked at low (3%) pO2. Similarly, biomineralization of long-term cell cultures was high under osteoblastic differentiation conditions in air but was undetectable at low (3%) pO2. In contrast, low pO2 upregulated mRNAs for OCT-4, REX-1, telomerase reverse transcriptase, and hypoxia-inducible factor-1 alpha, and increased the expression of SSEA-4 compared to air. Moreover, the expression of embryonic stem cell markers was sustained even under osteogenic culture conditions. Similar results were obtained using commercially available marrow stromal cells. We hypothesize a physiological scenario in which primitive MIAMI cells self-renew while localized to areas of low pO2 in the bone marrow, but tend to differentiate toward osteoblasts when they are located closer to blood vessels and exposed to higher pO2. Our results strongly suggest that maintaining developmentally primitive human cells in vitro at low pO2 would be more physiological and favor stemness over differentiation.

Alkaline Phosphatase↗

PubMed2004

The promise of human embryonic stem cells.

Pluripotency refers to the ability of a cell to give rise to cells that originate from all three germ layers. Among the available human pluripotent cells, human embryonic stem cells (hESCs) are considered to have the greatest probability for practical clinical application because of their simple propagation and stability in culture. Since their first derivation, issues concerning hESC maintenance and self-renewal have been widely addressed. The first part of this review presents the accumulated knowledge concerning the self-renewal of hESCs and discusses recent genetic profile data, which seem to shed light on hESC self-renewal and pluripotency mechanism. The second part deals with the regenerative potential of hESCs. Available lineage-specific differentiations of hESCs are presented, with detailed data on the ability of hESCs to differentiate into trophoblast cells, an observation that might broaden the definition of their developmental potential. Specific focus is given to vascular cell differentiation, including endothelial and smooth muscle cells. Transplantation limitations as well as current steps taken toward resolution conclude the review.

Blastocyst↗

PubMed2026

Functions and mechanisms of BRCA1 in early embryonic development.

Breast Cancer Gene 1 (BRCA1) is a critical regulator of genome integrity whose dysfunction greatly increases lifetime risk of breast and ovarian cancers. While BRCA1 has been extensively studied in the contexts of adult biology and cancer, its diverse functions, including homologous recombination-mediated DNA repair, cell cycle checkpoint activation, protein ubiquitination, and transcriptional regulation, have many underexplored implications. In early embryonic development, the maternal-to-zygotic transition (MZT) and subsequent developmental processes place extraordinary demands on DNA replication fidelity, cell cycle regulation, transcriptional activation, and chromatin remodeling. These critical processes overlap strikingly with canonical functions of BRCA1, yet its function in early development is poorly characterized. In this review, we investigate BRCA1 conservation across species and connect its well-established functions to findings from developmental studies to assess its role in development. We highlight evidence of BRCA1 mitigating genome integrity loss from diverse sources, maintaining the proliferative activity needed for successful germ layer formation and early tissue morphogenesis, and regulating transcription and epigenetic modifications. Together, this synthesis supports a model where BRCA1 acts as a multi-functional and dynamic regulator of early embryogenesis. Building on this, we propose outstanding questions that could further illuminate these developmental roles. Characterization of BRCA1 in early development may not only provide important insight into the origin and progression of cancer susceptibility but may also elucidate fundamental mechanisms shaping early development.

BRCA1 Protein↗