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Embryonic development of the CNS microvasculature in the mouse: new insights into the structural mechanisms of early angiogenesis.

Vascularization and expression of blood-brain barrier (bbb) associated morphological characteristics were studied by ultrastructural analysis during the embryonic development of the mouse CNS. At day 9 (E9) capillaries were only found in the perineural mesenchymal tissue exhibiting fenestrations and loosely attached pericytes. At E10 endothelial cells (EC) together with pericytes and other mesenchymal cells invaded the intraneural domain. The immigrating EC showed numerous protrusions and lost their fenestrations as soon as the new intraneural capillaries were formed. Intraneural EC showed prominent nuclei, many pinocytotic vesicles and junctional complexes which appeared tight in some places. These results indicate that from the first day of intraneural vascularization onwards, the morphological properties of the bbb are present in the early embryonic mouse cerebral cortex.

Animals↗

Role of nucleophosmin in embryonic development and tumorigenesis.

Nucleophosmin (also known as NPM, B23, NO38) is a nucleolar protein directly implicated in cancer pathogenesis, as the NPM1 gene is found mutated and rearranged in a number of haematological disorders. Furthermore, the region of chromosome 5 to which NPM1 maps is deleted in a proportion of de novo human myelodysplastic syndromes (MDS), and loss of chromosome 5 is extremely frequent in therapy-related MDS. NPM is a multifunctional protein, and its role in oncogenesis is controversial as NPM has been attributed with both oncogenic and tumour suppressive functions. To study the function of Npm in vivo, we generated a hypomorphic Npm1 mutant series (Npm1+/- < Npm1(hy/hy) < Npm1-/-) in mouse. Here we report that Npm is essential for embryonic development and the maintenance of genomic stability. Npm1-/- and Npm1(hy/hy) mutants have aberrant organogenesis and die between embryonic day E11.5 and E16.5 owing to severe anaemia resulting from defects in primitive haematopoiesis. We show that Npm1 inactivation leads to unrestricted centrosome duplication and genomic instability. We demonstrate that Npm is haploinsufficient in the control of genetic stability and that Npm1 heterozygosity accelerates oncogenesis both in vitro and in vivo. Notably, Npm1+/- mice develop a haematological syndrome with features of human MDS. Our findings uncover an essential developmental role for Npm and implicate its functional loss in tumorigenesis and MDS pathogenesis.

Animals↗

Embryonic development in the chick following exposure to ethanol, acetaldehyde and cyanamide.

The influence of cyanamide, an inhibitor of aldehyde dehydrogenase, on the embryopathic effects of ethanol and acetaldehyde, was investigated in the chick embryo. Both ethanol and cyanamide significantly increased embryonic mortality, but did not affect embryonic growth, compared to treatment with either ethanol or cyanamide. Acetaldehyde combined with cyanamide increased embryonic mortality and retarded embryonic growth. Cyanamide influence on embryonic development was minimal. The extent of acetaldehyde involvement in ethanol teratogenicity remains unclear from the present findings.

Acetaldehyde↗

Pairing SOX off: with partners in the regulation of embryonic development.

The SOX family of high-mobility group (HMG) domain proteins has recently been recognized as a key player in the regulation of embryonic development and in the determination of the cell fate. In the case of certain SOX proteins, they regulate the target genes by being paired off with specific partner factors. This partnering might allow SOX proteins to act in a cell-specific manner, which is key to their role in cell differentiation. The focus of this article is the mechanism of action of SOX proteins, in particular, how SOX proteins specifically pair off with respective partner factors and, as a consequence, select distinct sets of genes as their regulatory targets.

Animals↗

Influence of neostigmine treatment on embryonic development of acetylcholine receptors and neuromuscular junctions.

The postulated role of the acetylcholine receptor in the formation of neuromuscular synapses during the course of embryonic development was investigated in the superior oblique muscle of white Peking duck embryos. The possibility that the number of receptors could be experimentally lowered by chronic injections of the anticholinesterase agent, neostigmine methylsulfate, was determined using 125I-alpha-bungarotoxin. The total number of acetylcholine receptors on incubation day 12, 2 d subsequent to the onset of treatment, was reducted 45% as compared to saline-treated controls. A similar reduction in total receptor content (49%) was also observed on day 19. Radioautographic preparations showed that clusters of acetylcholine receptors were rare and that the grain density of extrajunctional receptors was also reduced. Hence, chronic treatment with neostigimine during development was observed to exert an effect on both the number and distribution of receptors in the developing superior oblique muscle. These changes occurred in the absence of any apparent effect on muscle differentiation in general. Myoblasts and myotubes were present on day 14 and further differentiated into myofibers by day 18 in both neostigmine and saline-treated muscles. The cytology of the develop;ing muscle cells also appeared normal. This is in contradistinction to the striking morphological changes that take place in adult mammalian and avian muscle after anticholinesterase treatment. More significantly, the decreased total receptor content and sparsity of clusters had no apparent effect on the formation of developing neuromuscular junctions at the electron microscopic level. The frequency of neuromuscular junctions in neostigmine-treated muscles was similar to that of the controls. It is concluded that acetylcholine receptor clusters are not required for the events leading to the morphological formation of neuromuscular junctions during in vivo development.

Animals↗

six-banded, a novel Drosophila gene, is expressed in 6 segmental stripes during embryonic development and in the eye imaginal disc.

We have characterised a Drosophila P-element enhancer detector insertion F125, which is expressed in the embryonic head and CNS as well as in various third instar imaginal discs. In an attempt to identify the gene with the equivalent expression pattern, we have characterised an adjacent gene. It encodes two novel conceptual proteins: Type I (1182 amino acids) and Type II, representing a shorter form of 774 amino acids truncated at both termini relative to Type I that is generated by alternative splicing. Based on its embryonic expression pattern, the gene was called six-banded (sba). Both splice forms are expressed in a unique embryonic pattern: initially as 6, then 12 stripes during early stages of embryonic development. Subsequently, expression is found in the developing trachae and during larval development is restricted to the eye imaginal disc where both transcripts are present immediately anterior to and behind the morphogenetic furrow. While sba expression in the eye antennal disc is mirrored by the expression of the adjacent F125 P-element, other patterns reported by this enhancer detector are not mimicked by the sba gene suggesting that the expression of the P-element represents a 'composite' of the effects of nearby enhancers.

Animals↗

Changes in proteins from yolk and in the activity of benzoyl-L-tyrosine ethyl ester hydrolase from the yolk sac membrane during embryonic development of the chicken.

The wet weight and nitrogen content in the water soluble fraction and granule fraction of chicken egg yolk changed during embryonic development. Rapid decreases between Days 14 and 16 were the largest changes observed. When protein components in the soluble and granule fractions of yolk from developing eggs were analyzed by polyacrylamide gel electrophoresis under denaturing conditions, the intensities of bands of some high molecular-weight proteins decreased during incubation, while smaller proteins increased. Most of such changes were observed after Day 16. The activities of proteinases and hydrolases assayed with protein substrates or synthetic substrates at a neutral pH were higher in the yolk sac membrane than in yolk itself. A benzoyl-L-tyrosine ethyl ester hydrolase showed the highest specific activity among the enzymes examined in yolk sac membrane homogenates. Its activity increased after Day 5 of embryogenesis, and became maximum on Days 14 to 20. The data suggest that some of the yolk proteins are degraded by yolk sac membrane enzymes during the latter half of the incubation period.

Animals↗

[Germ cells in the murine embryonic development].

This paper reviewed the recent progress of the origin, migration and proliferation, sex determination, and genomic modification of murine germ cells during its embryonic development. Murine germ cells originate from primordial germ cells at about 7-7.5dpc. Then PGCs migrated into germinal ridge at about 12.5dpc during which Steel/c-kit signal pathway plays important roles and stopped division at 13.5dpc. The sex of germ cells was mainly determined by the soma microenvironment in the gonad. And there are essential genes for sperm formation on the Y chromosome. The de novo methylation of murine germ cells was much later than soma cells and was completed at about 18.5dpc. The X chromosome reactivation of female germ cells was finished at about 14.5-15.5dpc which was independent of sexual differentiation of germinal ridge.

English Abstract↗

Genetic control of cuticle formation during embryonic development of Drosophila melanogaster.

The embryonic cuticle of Drosophila melanogaster is deposited by the epidermal epithelium during stage 16 of development. This tough, waterproof layer is essential for maintaining the structural integrity of the larval body. We have characterized mutations in a set of genes required for proper deposition and/or morphogenesis of the cuticle. Zygotic disruption of any one of these genes results in embryonic lethality. Mutant embryos are hyperactive within the eggshell, resulting in a high proportion reversed within the eggshell (the "retroactive" phenotype), and all show poor cuticle integrity when embryos are mechanically devitellinized. This last property results in embryonic cuticle preparations that appear grossly inflated compared to wild-type cuticles (the "blimp" phenotype). We find that one of these genes, krotzkopf verkehrt (kkv), encodes the Drosophila chitin synthase enzyme and that a closely linked gene, knickkopf (knk), encodes a novel protein that shows genetic interaction with the Drosophila E-cadherin, shotgun. We also demonstrate that two other known mutants, grainy head (grh) and retroactive (rtv), show the blimp phenotype when devitellinized, and we describe a new mutation, called zeppelin (zep), that shows the blimp phenotype but does not produce defects in the head cuticle as the other mutations do.

Amino Acid Sequence↗

Combined proton NMR imaging and spectral analysis of locust embryonic development.

Images and spectra of desert locust [Schistocerca gregaria (Forskal)] embryos were collected in vivo using a 4.7-tesla proton ((1)H) NMR imaging spectrometer. The 100 x 100 x 500 mum image resolution and 125-nl localized spectral volumes were obtained within minutes of each other. The dynamics of embryonic development were slow enough that the time delay between imaging and spectral measurements is negligible. Thus, image and spectral data correspond closely and approximate real-time observations of embryonic changes. The above procedure was applied every 12 hr during the entire course of development. This analytical approach demonstrates that imaging and localized spectroscopy can be used to visualize and assess changes in embryonic water and lipid content in concert with the development of a living embryo.

Journal Article↗

Expression of metallothionein genes during the post-embryonic development of Drosophila melanogaster.

Expression of the two Drosophila melanogaster metallothionein genes, Mtn and Mto, has been analyzed by in situ hybridization during post-embryonic development. Mtn and Mto transcripts were detected exclusively in the digestive tract of larvae, pupae and adults reared on standard medium. Mtn and Mto expression domains overlap, but each gene is also expressed at unique sites. Mtn mRNA levels are approximately 10 and 20 times higher than those of Mto in larvae and adults, respectively. Copper and cadmium ions strongly induce Mtn and Mto mRNA accumulation in the midgut. Zinc is a weaker inducer, acting only at high concentrations. Mtn gene expression is induced by these three metals in Malpighian tubules, while Mto gene expression in this organ is induced only by zinc. Iron is a poor inducer of metallothionein mRNA accumulation. Functions of MTN and MTO proteins in metal homeostasis and detoxification are considered.

Animals↗

Embryonic development after follicle culture is influenced by follicle-stimulating hormone isoelectric point range.

We evaluated the effects of follicular exposure in vitro to either of two mutually exclusive isoforms of FSH (least acidic and acid) on the subsequent capacity of oocytes for embryonic development. The effects of dose and follicle culture duration were examined. At the threshold dose (that required to produce antra) and at one subthreshold dose, the major difference between the two isoform fractions was the timing and effectiveness of acquisition of two-cell embryonic developmental capacity. With the least-acidic fraction, the highest rate of two-cell development (approximately 80%) occurred after 3 days of follicle culture only at the threshold dose (2.5 ng/ml). With the acid fraction, the highest two-cell rate (approximately 60%) occurred after 5 days of culture but at equivalent rates over a range of doses between 10 ng/ml and 100 ng/ml (threshold dose was 50 ng/ml). At threshold dose or below, the capacity for two-cell embryo production appeared not to be influenced by antral status for either isoform. At above threshold doses, the least-acidic fraction induced an increasing proportion of antral follicles with increasing dose, but this increase was associated with a progressive decrease in embryo production. This relationship was more extreme after longer culture and was due to degeneration of the cumulus-oocyte complex associated with apparently increased differentiation of the mural granulosa cells. The acid fraction was by comparison less bioactive and insensitive to overdosing. The broader isoform mix of the unfractionated FSH provided a measure of protection against overdosing characteristic of the acid fraction while retaining the capacity of the least-acidic fraction to induce antral formation at a low dose.

Animals↗

Insulin-like growth factor gene expression during rat embryonic development.

The function of insulin-like growth factors I and II (IGF-I and IGF-II) in embryogenesis is unknown. To investigate the ontogeny of IGF gene expression during mammalian development we used a highly sensitive and specific solution-hybridization assay to determine the steady state levels of IGF mRNAs during midgestation in the rat. IGF-I mRNA can be detected as early as day 11 of embryonic development and rises 8.6-fold over the ensuing 48 h. By contrast IGF-II mRNA is relatively constant over days 11-14 of gestation. These observations suggest that both IGFs may play important roles in early fetal development.

Animals↗

Cartilage-derived morphogenetic proteins. New members of the transforming growth factor-beta superfamily predominantly expressed in long bones during human embryonic development.

Partially purified extracts from newborn calf articular cartilage were found to induce cartilage and bone when subcutaneously implanted in rats. This activity showed characteristics of bone morphogenetic proteins (BMPs). Degenerate oligonucleotide primer sets derived from the highly conserved carboxyl-terminal region of the BMP family were designed and used in reverse transcription-polymerase chain reactions with poly(A)+ RNA from articular cartilage as template to determine which BMPs are produced by chondrocytes. Two novel members of the transforming growth factor-beta (TGF-beta) superfamily were identified and designated cartilage-derived morphogenetic protein-1 (CDMP-1) and -2 (CDMP-2). Their carboxyl-terminal TGF-beta domains are 82% identical, thus defining a novel subfamily most closely related to BMP-5, BMP-6, and osteogenic protein-1. Northern analyses showed that both genes are predominantly expressed in cartilaginous tissues. In situ hybridization and immunostaining of sections from human embryos showed that CDMP-1 was predominantly found at the stage of precartilaginous mesenchymal condensation and throughout the cartilaginous cores of the developing long bones, whereas CDMP-2 expression was restricted to the hypertrophic chondrocytes of ossifying long bone centers. Neither gene was detectable in the axial skeleton during human embryonic development. The cartilage-specific localization pattern of these novel TGF-beta superfamily members, which contrasts with the more ubiquitous presence of other BMP family members, suggests a potential role for these proteins in chondrocyte differentiation and growth of long bones.

Amino Acid Sequence↗

[Histochemistry and electron microscopy of the embryonal development of the golden hamster kidney: study of the interstitial cells].

The structure and the origin of the interstitial cells in the kidney of the golden Hamster during its embryonic development was studied. For this purpose kidneys from 11, 13, 15 and 17 day-old embryos were used. The histochemical observations as well as the electron microscopy demonstrated that the interstitial cells have a metanephric origin and are embryonically similar to the cellular elements that differentiate the primordial renal vesicles and later organise the epithelial portion of the nephrons. These cells seem to take part in the formation of the basal membranes during tubular genesis.

Acid Phosphatase↗

Relationships between follicular fluid steroid hormone concentrations, oocyte maturity, in vitro fertilization and embryonic development in the rhesus monkey.

Oocytes and matched samples of follicular fluid (FF) were obtained from 70 follicles of five rhesus monkeys stimulated with either pregnant mare serum gonadotropin or human menopausal gonadotropin. Follicular aspiration was performed 30-32 h after human chorionic gonadotropin administration. The concentrations of estradiol (E2), progesterone (P), testosterone (T), and dihydrotestosterone (DHT) in FF were measured. Twenty-six percent of oocytes were classified as mature (M), 41% matured in vitro (Miv), 13% were dysmature, and 20% atretic. M oocytes were associated with significantly higher levels of P and a higher P:E2 ratio. There were no differences in hormone levels associated with fertilized and nonfertilized oocytes. Thirty-five embryos developed to the six- to eight-cell stage in vitro, of which 13 exhibited optimal cleavage rates. Significantly lower levels of E2 and higher P:E2 ratios were associated with the more rapidly cleaving embryos. Proportionally more embryos showing optimal cleavage rates developed from M compared to Miv oocytes, and only embryos derived from M oocytes developed to blastocysts in culture. Optimal cleavage rates to the six- to eight-cell stage in vitro, rather than fertilization rates, are a better indicator of (subsequent) developmental capacity, and, in this study, embryonic development was closely associated with the maturity of the oocyte at recovery.

Animals↗

Gliogenesis and ependymogenesis during embryonic development of the rat. An autoradiographic study.

With the aid of [3H]thymidine autoradiography gliogenesis and ependymogenesis were studied in the brain of the rat during embryonic development. Gliogenesis was found to begin on day 17 of gestation in the caudal regions of the brain stem, and to spread rostrally. On days 20 and 21 of gestation gliogenesis reached a peak, and then declined. Ependymogenesis began earlier and showed the following pattern: day 14 of gestation in the 4th ventricle and cerebral aqueduct, day 15 in the 3rd ventricle, and day 17 in the lateral ventricles reaching a peak on different days in different sites. Both gliogenesis and ependymogensis continued up to the last day of gestation, day 22. Issues pertaining to gliogenesis and the formation of glioblasts, and the relationship between gliogenesis and ependymogenesis are discussed.

Animals↗