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PubMed1988

Ontogeny of calcitonin gene-related peptide-immunoreactive structures in the rat forebrain and diencephalon.

Ontogeny of the calcitonin gene-related peptide (CGRP)-containing structures was investigated in the forebrain and diencephalon of the rat by means of indirect immunofluorescence. CGRP-like immunoreactive (CGRP-IR) fibers first appeared at gestational day 18 (E18) in the primordia of the bed nucleus of the stria terminalis, central amygdaloid nucleus and posterolateral portion of the lateral hypothalamic nucleus. CGRP-IR fibers gradually increased in immunoreactivity and in number in these nuclei toward birth. A pronounced increase of CGRP-like immunoreactive fibers was found after birth in these nuclei and made very dense plexuses of fibers in adult animals. A few CGRP-IR fibers first appeared at the perinatal stage in the lateral septum, the most caudal portion of the caudate-putamen, and ventromedial and ventroposterior thalamic nuclei. Then the immunoreactive fibers increased and dense plexuses were present in these brain areas of the adult. A small number of immunoreactive fibers appeared at postnatal day 4-7 (P4-7) in the caudal insular cortex and ventromedial hypothalamic nucleus. A significant number of fibers was present in these areas of the adult. On the other hand, CGRP-like immunoreactivity in cell bodies was first detected with faint immunofluorescence at P4 in the anteromedial parts of the lateral hypothalamic area and ventromedial and ventroposterior thalamic nuclei. Thereafter CGRP-IR cells increased in immunoreactivity, and a significant number of cells was noted in these nuclei of the adult. In addition, CGRP-IR cells are identified in the medial amygdaloid nucleus, and medial preoptic area at P14. In conclusion, in the forebrain and diencephalon (1) CGRP-like immunoreactivity appears in fibers earlier than in cell bodies, suggesting that at least some of immunoreactive fibers originate from the lower brainstem, and (2) high density of CGRP-IR structures present in the brain at embryonic and early postnatal stages may indicate that CGRP could be an important factor involved in the developmental organization of the central nervous system.

Aging↗
PubMed1994

cDNA sequence of zebrafish (Brachydanio rerio) translation elongation factor-1 alpha: molecular phylogeny of eukaryotes based on elongation factor-1 alpha protein sequences.

We have isolated and determined the nucleotide sequence of a cDNA clone containing the complete coding region for elongation factor-1 alpha (EF-1 alpha) from an embryonic zebrafish cDNA library. A secondary structure model based on all known EF-1 alpha and EF-Tu protein sequences is presented and the presence of conserved putative protein kinase C phosphorylation sites in loop regions of eukaryotic EF-1 alpha is demonstrated. Using distance matrix and maximum parsimony methods we constructed multi-kingdom phylogenetic trees containing 22 different eukaryotic sequences. Strikingly, both tree constructions show Fungi to be the closest relative of Animalia among eukaryotic kingdoms. A 12 amino acid stretch present in all animal and fungal sequences known to date was found to be absent from all plant, protist an archaebacterial EF-1 alpha sequences suggesting that this sequence was inserted following the separation of plants from the lineage leading to fungi and animals. In contrast to our results, molecular phylogenies based on small subunit ribosomal RNA sequences as well as other protein sequences have failed to yield consistent results regarding the branching order among the kingdoms Plantae, Fungi and Animalia. The slow evolutionary rate and universal occurrence of EF-1 alpha (EF-Tu in eubacteria) makes this protein a particularly interesting tool for probing distant evolutionary relationships.

Animals↗
PubMed1993

The development and function of the Drosophila CNS midline cells.

1. The midline cells of the Drosophila embryonic CNS comprise a discrete neuroanatomical structure consisting of a small subset of neurons and glia. 2. Developmental commitment of the CNS midline cells requires the action of dorsal/ventral patterning genes. 3. The single-minded gene encodes a basic-helix-loop-helix transcription factor and acts as a master regulator for the CNS midline lineage. 4. A number of different transcription factors and proteins involved in cell-cell interactions are necessary for the differentiation of midline neurons and glia. 5. CNS midline cells have important functions in the formation of the ventral epidermis and axon commissures.

Animals↗
PubMed1994

Modulation of wingless signaling by Notch in Drosophila.

Extensive genetic and molecular analyses indicate that Notch acts as a transmembrane receptor in an evolutionarily conserved cell interaction mechanism that appears to control a common step in the progression of an uncommitted cell towards the differentiated state. In Drosophila, Notch mutations were shown to affect the development of a broad spectrum of tissues, including the wing. We found that mutations in the segment polarity gene wingless are capable of acting as dominant enhancers of notchoid, a recessive Notch allele affecting the wing. The Wingless protein is homologous to the mammalian proto-oncoprotein Wnt-1 and is thought to act as the signal in a cell interaction mechanism that specifies differentiation of the embryonic epidermis as well as imaginal structures such as the wing. Although some components of the Wingless signal transduction pathway have been identified, the receptor for Wingless remains elusive. This genetic link between the Wingless and Notch pathways has been further examined by determining the relative expression patterns and subcellular localization of Notch and Wingless in mutant and wild-type backgrounds. We find that Notch is necessary for the implementation of the Wingless signal in specifying normal wing development. We discuss the possibility that Notch is directly involved in the reception of Wingless in the light of current models for the developmental action of Notch.

Animals↗
PubMed2006

Effects of cyanobacterium Fischerella ambigua isolates and cell free culture media on zebrafish (Danio rerio) embryo development.

The toxic effects of several species of fresh water cyanobacteria, notably Microcystis species and associated toxins, the microcystins, Anabaena species (anatoxin), Nodularia sp. (nodularin), and Cylindrospermopsis raciborskii (cylindrospermopsin), are well known. Little, however, is known about the effects of secondary metabolites other than alkaloids. Early life stage tests with zebrafish (Danio rerio) were used to detect bioactive properties of compounds released by healthy cyanobacteria (Fischerella ambigua), particularly on the early developmental stages of fish. This approach, using F. ambigua is probably most valuable as it shows the toxicity of healthy growing cyanobacteria. The effects of cyanobacterial secondary metabolites on the embryonic stages of fish are of considerable interest as many aquatic creatures, particularly fish, are unable to avoid the potential toxins that may be released by undesirable algal blooms or as a result of allelopathic effects. In the current study, the zebrafish (D. rerio) was used as a model experimental system to investigate the effects of ambigols A and C, tjipanazole D and C, 2,4-dichlorobenzoic acid, cell free culture media, and media extracts of a terrestrial/fresh water strain of the cyanobacterium F. ambigua on embryo development. Fish embryo tests performed with the cell free culture medium showed that after 3h of exposure to undiluted culture medium all fish embryos died. At a tenfold dilution the process of epiboly (formation of the gastrula) was retarded in all embryos, lesions were observed, and their general development was significantly arrested, finally followed by death. The same tests performed with extracts (dichloromethane, n-butanol, and residual cell free culture medium) of the cell free culture medium, ambigol A, ambigol C, 2,4-dichlorobenzoic acid and tjipanazole D showed only ambigol A to have an influence on zebrafish development at concentrations>or=1 mg/l (2.06 microM). After 55 h all embryos showed pectoral oedema, irregularly shaped fin folds, bent tails, and unusual circular neoplasms in the dorsal tail fin fold. Due to the high concentration of ambigol A used in this assay these effects were considered to be of minor importance when compared to those of the culture medium.

Animals↗
PubMed2004

Thrombospondin 1: a multifunctional protein implicated in the regulation of tumor growth.

Thrombospondins belong to a family of extracellular matrix (ECM) proteins widely found from embryonic to adult tissues. The modular structure of thrombospondins contains a series of peptide sequences implicated in a multiplicity of biological functions. Extracellular matrix undergoes important alterations under proteolysis that occurs in pathological processes like tumorigenesis. An elevated secretion of thrombospondin 1 (TSP1) is often observed in tumors and is sometimes considered as a predictive factor. However, the role of TSP1 in cancer progression remains controversial and must be carefully apprehended. The regulation of cell adhesion, proliferation, apoptosis by TSP1 is examined in the present review and it is clear from the literature and from our investigations that TSP1 presents both stimulatory and inhibitory effects. The exposition of cryptic sites upon conformational changes can partially explain this contradiction. More interestingly, the analysis of TSP1-directed intracellular signaling pathways activated through specific receptors or supramolecular receptors docking systems may be useful to discriminate the precise function of TSP1 in tumor progression. The central role played by TSP1 in the control of matrix-degrading enzyme activation and catabolism reveals attractive tracks of research and highlights the involvement of the lipoprotein receptor-related protein (LRP) receptor in these events. Therefore, TSP1-derived peptides constitute a source of potentially active matrikins which could provide essential tools in cancer therapy.

Apoptosis↗
PubMed2025

Mouse totipotent blastomere-like cells model embryogenesis from zygotic genome activation to post implantation.

Embryo development begins with zygotic genome activation (ZGA), eventually generating blastocysts for implantation. However, in vitro systems modeling the pre-implantation development are still absent and challenging. Here, we used mouse totipotent blastomere-like cells (TBLCs) to develop spontaneous differentiation and blastoid formation systems, respectively. We found Wnt signaling enabled the rapid expansion of TBLCs and the optimization of their culture medium. We successfully developed a TBLC-spontaneous differentiation system in which mouse TBLCs (mTBLCs) firstly converted into two types of ZGA-like cells (ZLCs) distinguished by Zscan4 expression. Surprisingly, Zscan4-, but not Zscan4+, ZLCs further passed through intermediate 4-cell and then 8-cell/morula stages to produce epiblast, primitive endoderm, and trophectoderm lineages. Significantly, single TBLCs underwent expansion, compaction, and polarization to efficiently generate blastocyst-like structures and even post-implantation egg-cylinder-like structures. Conclusively, we established TBLC-based differentiation and embryo-like structure formation systems to model early embryonic development, offering criteria for evaluating and understanding totipotency.

Animals↗
PubMed2002

Muscarinic receptor protein expression in the ocular tissues of the chick during normal and myopic eye development.

Muscarinic receptor signalling has been implicated in both the embryonic and postnatal development of ocular structures as well as in myopic eye growth. A radioligand binding assay was used to determine whether changes in muscarinic receptor density and/or affinity occurred in the chick retina, choroid and sclera during early post-hatching development or with the induction of myopia. Specific receptor binding sites were saturable with increasing concentrations of the muscarinic receptor ligand [3H]N-methyl-scopolamine in the retina and choroid but not in the sclera. In normal eyes, binding density in the retina was not altered from age P5 to P10 (447+/-14 vs. 411+/-13 fmol/mg of protein, P=0.07). However, in the choroid, the number of receptor binding sites significantly increased between P5 and P10 (637+/-39 vs. 1125+/-121 fmol/mg of protein, P<0.01). Binding affinity (K(D)) was not altered with age in either the retina or choroid. Myopia was induced in chicks by deprivation of form vision, using translucent diffusers, from age P3. Despite the induction of significant degrees of ocular elongation and myopia at P5 (-8.7+/-0.3 D, P<0.01) and P10 (-22.5+/-1.3 D, P<0.01), neither muscarinic receptor density nor affinity were altered in the retina or choroid of myopic eyes. These findings indicate that regulation of muscarinic receptor numbers in the chick choroid is occurring in normal post-hatching development of this tissue. However, myopic eye enlargement was not associated with changes in muscarinic receptor protein expression in the chick retina and choroid.

Aging↗
PubMed1996

Early olfactory fiber projections and cell migration into the rat telencephalon.

The formation and development of primary olfactory axons was studied in the rat embryo using acetylcholinesterase histochemistry, immunocytochemistry for neuron-specific beta-tubulin (TuJ1) and growth associated protein 43 (GAP43), and a fluorescent tracer DiI. Olfactory axons extend from the olfactory receptor neurons localized in the olfactory epithelium. These fibers grow to reach and enter the olfactory bulbs, where they form the first relay and integrative synaptic station in the olfactory system: the olfactory glomerulus. In this communication we address the development of primary olfactory fibers: first from the olfactory placode and later from the olfactory epithelium. Olfactory fibers enter the olfactory bulbs apparently in a disordered manner but soon arrange themselves in hook shaped aggregates of fibers, with many boutons (immature synaptic terminals), to form the glomeruli. We detected this kind of structure for the first time at embryonic day 16. The olfactory receptor cells are usually anchored in the basal lamina of the olfactory epithelium but some of them, after reaching their targets, lose their epithelial attachment, leave the olfactory epithelium and migrate to and enter the olfactory bulbs. The traffic of cells between the olfactory epithelium and the brain lasts late into embryonic development. We describe four types of migratory mechanism used by different populations of cells to reach their targets in the telencephalic vesicle and propose the existence of migrating cells that enter the telencephalon. These data were corroborated by injections into the olfactory epithelium a of murine retrovirus carrying the Escherichia coli lac-Z gene.

Acetylcholinesterase↗
PubMed1997

Conservation of BMP signaling in zebrafish mesoderm patterning.

To investigate the conservation of mechanisms for mesodermal patterning between zebrafish and Xenopus, we isolated two cDNA clones encoding bone morphogenetic protein (BMP)-related proteins from a zebrafish cDNA library. Based on their predicted amino acid sequences, these two clones were designated as zbmp-2 and zbmp-4. Whole-mount in situ hybridization analysis revealed that in gastrula embryo, both genes were localized in the ventral part of the embryo, consistent with the proposed function of Xenopus BMP-4 in ventral mesoderm specification. zbmp-4 expression, however, was also seen in the embryonic shield, the most dorsal mesodermal structure. To examine the ability of zbmp-2 to ventralize mesoderm, we injected synthetic mRNA into zebrafish embryos and found that overexpression of this gene eliminated dorsal structures including notochord at both morphological and molecular level. In contrast, expression of ventral marker gene eve1 was expanded to the dorsal side. These effects are analogous to the ventralization of embryos caused by ectopic xBMP-4 expression. Taken together, one may conclude that the developmental mechanisms for mesodermal patterning regulated by BMPs are evolutionarily conserved between amphibians and teleosts.

Amino Acid Sequence↗
PubMed1996

Pulsed electromagnetic fields influence hyaline cartilage extracellular matrix composition without affecting molecular structure.

Pulsed electromagnetic fields (PEMF) influence the extracellular matrix metabolism of a diverse range of skeletal tissues. This study focuses upon the effect of PEMF on the composition and molecular structure of cartilage proteoglycans. Sixteen-day-old embryonic chick sterna were explanted to culture and exposed to a PEMF for 3 h/day for 48 h. PEMF treatment did not affect the DNA content of explants but stimulated elevation of glycosaminoglycan content in the explant and conserved the tissue's histological integrity. The glycosaminoglycans in sterna exposed to PEMF were indistinguishable from those in controls in their composition of chondroitin sulfate resulting from chondroitinase ABC digestion. Specific examination with [35S]-sulfate labels showed that PEMF treatment significantly suppressed both the degradation of pre-existing glycosaminoglycans biosynthetically labeled in ovo and the synthesis of new [35S]-sulfated glycosaminoglycans. The average size and aggregating ability of pre-existing and newly synthesized [35S]-sulfated proteoglycans extracted with 4 M guanidinium chloride from PEMF-treated cartilage explants were identical to controls. The chain length and degree of sulfation of [35S]-sulfated glycosaminoglycans also were identical in control and PEMF-treated cultures. PEMF treatment also reduced the amount of both unlabeled glycosaminoglycans and labeled pre-existing and newly synthesized [35S]-sulfated glycosaminoglycans recovered from the nutrient media. [35S]-Sulfated proteoglycans released to the media of both control and PEMF-treated cultures were mostly degradation products although their glycosaminoglycan chain size was unchanged. These results demonstrate that exposure of embryonic chick cartilage explants to PEMF for 3 h/day maintains a balanced proteoglycan composition by down-regulating its turnover without affecting either molecular structure or function.

Animals↗
PubMed1987

The organization of the oocyte and embryogenesis in twinning and fusion malformations.

Certain congenital malformations occur in excess among twins and also among first-degree relatives of twins. In the general population, these anomalies are familially associated with each other, and, like twinning, familially associated with unusual brain function asymmetry. They affect structures built by fusion of bilateral embryonic halves and remodeled under major influence of neural crest mesenchyme. This conjunction of associations suggests that twinning, symmetry development, and this group of malformations might share causal elements at least some of which are heritable. The problem here is the absence of zygosity differences in these relationships, because of which they cannot be explained within the biology of twinning as it has been understood. A potential resolution is offered by way of a mechanism common to MZ and DZ twinning, involving a relationship between oocyte organization and the determination of body symmetries.

Congenital Abnormalities↗
PubMed2002

Age-related changes of thymalin content in human epidermis.

Immunomorphological analysis revealed the presence of thymalin in human epidermis and in fetal reticuloepithelium. These structures are developed from the common embryonic primordium ectoderm. In embryos and adult humans thymalin is present only in young epidermal cells, which undergo age-related involution. By the age of 70 years, the layer of thymalin-containing cells looks thinned and discontinuous. The content of thymalin, a thymic factor, decreases with age.

Adult↗
PubMed1984

Identification and characterization of insulin receptors on foetal-mouse brain-cortical cells.

The occurrence of insulin receptors was investigated in freshly dissociated brain-cortical cells from mouse embryos. By analogy with classical insulin-binding cell types, binding of 125I-insulin to foetal brain-cortical cells was time- and pH-dependent, only partially reversible, and competed for by unlabelled insulin and closely related peptides. Desalanine-desasparagine-insulin, pig proinsulin, hagfish insulin and turkey insulin were respectively 2%, 4%, 2% and 200% as potent as bovine insulin in inhibiting 125I-insulin binding to brain-cortical cells, which corresponds to their relative biological potencies in classical insulin-target cells; no competition was observed with glucagon and nerve growth factor, even at high concentrations. Scatchard analysis of competitive-binding data resulted in curvilinear plots with a high-affinity binding of Ka = 3.6 X 10(8) M-1. Insulin binding to foetal brain-cortical cells differed, however, in two distinct aspects from that to classical insulin-binding cell types. Firstly, dilution of 125I-insulin-bound cells in the presence of unlabelled insulin did not accelerate dissociation of the labelled hormone. Secondly, exposure of brain-cortical cells to insulin before the binding assay enhanced insulin binding, suggesting up-regulation of insulin receptors in response to insulin. In conclusion, foetal-mouse brain-cortical cells bear specific binding sites for insulin. Their insulin receptor shows a marked specificity and affinity for insulin, but differs in at least two properties from most classical insulin receptors. These differences in hormone-receptor interaction could reflect structural differences between insulin receptors on embryonic and differentiated cells.

Animals↗
PubMed1989

High-resolution autoradiographic studies of comparative nucleologenesis and genome reactivation during early embryogenesis in pig, man and cattle.

Using high-resolution autoradiography, simultaneous studies of ultrastructure and nucleic acid dynamics were performed during nucleologenesis in early porcine, human and bovine embryos. In contrast to the early genome activation known to occur during the second cell cycle in the mouse, the onset of rRNA synthesis detected by (5-3H) uridine incorporation in the nucleolar compartment is delayed by cleavage of one cell cycle (to the third cell cycle) in the early pig embryo and by two cell cycles (to the fourth cell cycle) in human and cattle embryos. Extranucleolar RNA synthesis, as detected by nucleoplasm labelling, generally started shortly before rRNA synthesis. The timing of nucleolar labelling was well correlated with the penetration of embryonic DNA into the nucleolus precursor body and with nucleolus structure differentiation. Ultrastructural and/or autoradiographic techniques are suggested for the study of the onset of embryonic transcription, e.g. in embryos "reconstructed" by micromanipulation.

Animals↗
PubMed2002

[Intestinal polyp of the umbilical cord].

The morphology, pathogenesis, complications and differential diagnosis of an intestinal polyp of the umbilical cord are presented. The polyp were detected postnatal on the umbilical cord in an healthy male newborn. The presents of intestinal tissue upon the umbilical cord ist possible about the persistence from remnants of the ductus omphalomesentericus with prolapse and differentiation of the intestinal cells. The ductus omphalomesentericus is a tubular structure, a communication between the developing embryonic gut and the yolk sac, forming during the early embryonic life. Obliteration of the omphalomesenteric duct is usually complete by the 10(th) week of gestation. Various portions of the duct may persist, however, giving rise to polyps, fistulas or cysts of the umbilical cord with potentially dangerous clinical consequences. Other tumors of the umbilical cord are myxoma, angioma and teratoma are differential diagnosis.

Adult↗
PubMed2006

The role of the proline-rich domain of Ssdp1 in the modular architecture of the vertebrate head organizer.

Lim1, Ssdp1, and Ldb1 proteins are components of the Ldb1-associated transcriptional complex, which is important in the head-organizing activity during early mouse development. Depletion of each individual protein alone causes a headless phenotype. To explore in more detail the modular architecture of the complex, we have generated two different gene-trapped mouse lines that express truncated forms of Ssdp1. Embryos derived from the gene-trapped line that encodes a truncated Ssdp1 lacking the proline-rich sequence exhibit a lethal abnormal head-development phenotype, resembling mouse embryos deficient for Lim1, Ssdp1, or Otx2 genes. Embryos derived from the second gene-trapped line, in which most of the proline-rich domain of Ssdp1 is retained, did not show abnormalities in head development. Our data demonstrate that components of the Ldb1-dependent module can be subdivided further into discrete functional domains and that the proline-rich stretch of Ssdp1 is critical for embryonic head development. Furthermore, phylogenetic comparisons revealed that in Caenorhabditis elegans, a similar proline-rich sequence is absent in Ssdp but present in Ldb1. We conclude that although the overall architecture of the Ldb1-dependent module has been preserved, the genetic specification of its individual components has diversified during evolution, without compromising the function of the module.

Amino Acid Sequence↗
PubMed2001

Vascular patterning defects associated with expression of activated Notch4 in embryonic endothelium.

Notch proteins function as receptors for membrane-bound ligands (Jagged and Delta-like) to regulate cell-fate determination. We have investigated the role of Notch signaling in embryonic endothelium of the mouse by expressing an activated form of the Notch4 protein in vasculature under the regulation of the Flk1 (VEGFR) locus. Expression of activated Notch4 results in a growth and developmental delay and embryonic lethality at about 10 days postcoitum. The extent of the developing vasculature in mutant embryos was restricted, fewer small vessels were seen, and vascular networks were disorganized. The brain periphery of mutant embryos contained large dilated vessels with evidence of compromised vessel-wall integrity and large areas of necrosis; yolk-sac vasculature was abnormal. Expression of an activated form of Notch4 in embryonic vasculature leads to abnormal vessel structure and patterning, implicating the Notch pathway in phases of vascular development associated with vessel patterning and remodeling.

Animals↗