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cDNA sequence analysis and expression of the expression of the ribosomal protein S24 during oogenesis and embryonic development of the sea urchin Paracentrotus lividus.

A gene for the Paracentrotus lividus ribosomal protein S24, called P1-S24, has been isolated and sequenced. Ribosomal protein P1-S24 consists 130 amino acids and has a molecular weight of 14869 Da. Sequence analysis shows a high percentage (90%) identity with the corresponding gene of Strongylocentrotus purpuratus. Hybridization of the cDNA to digested sperm DNA suggests that P1-S24 is represented in no more than two copies. Studies of the temporal expression of P1-S24 gene indicate a good correlation between this and the expression of the rRNA genes both during oogenesis and embryonic development.

Amino Acid Sequence↗

Signals regulating tendon formation during chick embryonic development.

Tendons are collagen-rich structures that link muscle to cartilage. By using quail-chick chimeras, it has been shown that tendon and cartilage cells originate from the same mesodermic compartment, which is distinct from that giving rise to muscle cells. Axial tendons originate from the sclerotomal compartment, and limb tendons originate from the lateral plate, whereas axial and limb muscles derive from dermomyotomes. Despite these different embryologic origins, muscle and tendon morphogenesis occurs in close spatial and temporal association. Facilitated by the distinct embryologic origin of myogenic and tendon cells, surgical studies in the avian embryo have highlighted interactions between tendons and muscles, during embryonic development. However, these interactions seem to differ between axial and limb levels. The molecular mechanisms underlying muscle and tendon interactions have been shown recently to involve different members of the fibroblast growth factor family. This review covers the available data on the early steps of tendon formation in the limb and along the primary axis. The relationship with muscle morphogenesis will be highlighted.

Animals↗

Expression of polypeptide variants of receptor-type protein tyrosine phosphatase beta: the secreted form, phosphacan, increases dramatically during embryonic development and modulates glial cell behavior in vitro.

Glial cells express three splicing variants of a receptor-type protein tyrosine phosphatase called RPTP beta. Two are receptor forms that differ in a large extracellular domain. The third is a secreted proteoglycan called phosphacan that lacks the cytoplasmic phosphatase domains. We have now identified, by immunoblotting, proteins corresponding to these three forms of RPTP beta in rat C6 glioma cells and brain. The short receptor form is much more prevalent than the full-length receptor in C6 glioma cells. Phosphacan is much more abundant than either of the receptor forms in rat brain, and its expression increases progressively during embryonic development, while the receptor forms show only moderate changes. In contrast to the long form and phosphacan that were detected as proteoglycans, the short receptor form, lacking the large alternatively spliced domain, was not detected as a chondroitin sulfate proteoglycan. We recently showed that phosphacan binds to the neuron-glia cell adhesion molecule, Ng-CAM, and we now report that glia expressing RPTP beta adhere and extend processes on substrates coated with Ng-CAM. After one day in culture, however, the glia retract their processes and often lift off the substrate. Conditioned medium from glial cells, which contains large amounts of phosphacan, inhibits glial adhesion to Ng-CAM, and depletion of phosphacan from the conditioned medium by immunoadsorption reduces the inhibitory activity. The results show that phosphacan increases dramatically during development, and indicate that secreted forms of RPTP beta can modulate glial cell adhesion and behavior.

Animals↗

Neurotrophins are not required for normal embryonic development of olfactory neurons.

Neurons of the vertebrate olfactory epithelium (OE) regenerate continuously throughout life. The capacity of these neurons to regenerate and make new and precise synaptic connections in the olfactory bulb provides a useful model to study factors that may control or mediate neuronal regeneration. Expression and in vitro studies have suggested potential roles for the neurotrophins in the olfactory system. To directly examine whether neurotrophins are required for olfactory neuron development, we characterized in vivo the role of the neurotrophins in the primary olfactory system. For this, we generated mutant mice for TrkA, TrkB, TrkC, and also for BDNF and NT3 together with P2-IRES-tau-LacZ trangenic mice. Histochemical staining for beta-galactosidase at birth allowed in vivo analysis of the P2 subpopulation of olfactory neurons as well as their projections to the olfactory bulb. Our data indicate that Trk signaling is not required for normal embryonic development of the olfactory system.

Animals↗

Autonomic regulation of calcium cycling in developing embryonic mouse hearts.

In the present study, we combined optical Ca(2+) imaging with immunocytochemistry studies to characterize autonomic regulation of Ca(2+) cycling during early development in isolated embryonic mouse hearts. At embryonic days 9.5-11.5 (E9.5-E11.5), the Ca(2+) transient originated in the superior portion of the right atrium, propagated rapidly through both atria, slowly through the atrio-ventricular (AV) ring, and rapidly through both ventricles. Isoproterenol (ISO) significantly increased heart rate, increased Ca(2+) transient amplitude, rate of rise (RR) and a rate of decay, and shortened AV conduction time, indicating the presence of functional beta-adrenergic receptors. The muscarinic agonist carbachol (CCh) had no effects until 1 day later at E10.5. Both beta1-adrenergic and M2 muscarinic receptors were detected in ventricular muscle sections by immunochemistry at E10.5. Growing nerves, labeled using growth-associated protein 43 antibodies, were detected at the E14.5 stage, but not at E10.5, whereas mature sympathetic nerves, detected by tyrosine hydroxylase (TH) labeling, were not yet present at E14.5. These results demonstrate that functional regulation of Ca(2+) cycling by beta-adrenergic receptors occurs earliest in developing embryonic mouse hearts, followed a day later by muscarinic receptor responsiveness, with autonomic innervation developing later. These results define the functional and structural sequence of autonomic regulation of Ca(2+) transient in the embryonic mouse heart.

Age Factors↗

The zebrafish BMP4 gene: sequence analysis and expression pattern during embryonic development.

We have isolated zebrafish BMP4 gene from a zebrafish genomic DNA library. The size of the isolated BMP4 gene was approximately 14.9 kb. The isolated gene contained two exons which formed the complete coding region together with part of the 3'-noncoding region. The deduced BMP4 protein sequence contained 400 amino acids. Sequence comparison showed that it shared 73% amino acid sequence identity with that of human and mouse BMP4. An intron with a size of 8,963 bp was present between two coding exons. Danio retroposon A (DANA)-like retroposon was located in the intron. It contained four conserved boxes and was flanked by a pair of direct repeats of 9 nucleotide sequence (GTTTTAATA). During embryonic development of the zebrafish, a 3.8-kb BMP4 mRNA was detected from gastrula stage up to a month-old hatching larvae via Northern blot analysis. In addition, the use of reverse transcription polymerase chain reaction further demonstrated the presence of BMP4 mRNA in both the early developmental stages (i.e., cleavage and blastula) and in adult fish. Developmental expression of BMP4 protein was also analyzed. Trace amounts of an 18-kD protein were detected at pharyngula stage, while the production increased from hatching larvae to adult fish. In adult fish, the expression of BMP4 mRNA was observed in brain, heart, digestive tracts, testes, and jaw. The results suggest that the zebrafish BMP4 gene may play important roles during zebrafish development.

Amino Acid Sequence↗

Reconstitution of membranes and embryonic development in dissociated blastula cells of the sea urchin by reinsertion of aggregation-promoting membrane proteins extracted with butanol.

Blastula embryos of the sea urchin Paracentrotus lividus, when dissociated into single cells by exposure to Ca2+- and Mg2+-free sea water, reassociate spontaneously to form aggregates capable of development to the final larval form (pluteus). This aggregation is prevented by Fab fragments obtained by immunization with purified membranes from blastula embryos. The inhibition was reversed by soluble proteins extracted with butanol from purified membranes or from intact cells. These extracts also strongly stimulated the rate of reaggregation of dissociated cells in the absence of Fab fragments. Exposure of dissociated cells to 2.5% (vol/vol) butanol removed completely the protein(s) responsible for reaggregation of the cells without impairing their viability. Reaggregation and embryonic development were completely restored to the extracted cells by readdition of the proteins extracted from either membranes or cells. Extracted cells from Paracentrotus could be reconstituted with proteins from Arbacia.

Animals↗

The involvement of a conserved family of RNA binding proteins in embryonic development and carcinogenesis.

Vg1 RBP is a member of a family of highly conserved proteins that appear to be involved in RNA localization, stability, and/or translational control in a wide variety of cell types and organisms. Over the last few years, the human homologs of these proteins have been found to be overexpressed in an increasing number of different kinds of cancers. Although the role of these proteins in neoplasia is not understood, results from several labs, including our own, are beginning to suggest that many of these proteins may be important in cell motility, a necessary requirement for metastasis. This paper will review these data and suggest a model for the role of Vg1 RBP and its homologs in embryonic development and carcinogenesis.

Animals↗

Organization versus activation: the role of endocrine-disrupting contaminants (EDCs) during embryonic development in wildlife.

Many environmental contaminants disrupt the vertebrate endocrine system. Although they may be no more sensitive to endocrine-disrupting contaminants (EDCs) than other vertebrates, reptiles are good sentinels of exposure to EDCs due to the lability in their sex determination. This is exemplified by a study of alligators at Lake Apopka, Florida, showing that EDCs have altered the balance of reproductive hormones resulting in reproductive dysfunction. Such alterations may be activationally or organizationally induced. Much research emphasizes the former, but a complete understanding of the influence of EDCs in nature can be generated only after consideration of both activational and organizational alterations. The organizational model suggests that a small quantity of an EDC, administered during a specific period of embryonic development, can permanently modify the organization of the reproductive, immune, and nervous systems. Additionally, this model helps explain evolutionary adaptations to naturally occurring estrogenic compounds, such as phytoestrogens.

Animals↗

The effect of female antisperm antibodies on in vitro fertilization, early embryonic development, and pregnancy outcome.

STUDY OBJECTIVE: To evaluate the extent to which human in vitro fertilization-embryo transfer (IVF-ET) alleviates immunological infertility. DESIGN: Retrospective. SETTING: In vitro fertilization program. PATIENTS: Thirty-three patients with positive antisperm antibodies undergoing 50 cycles of IVF-ET in which maternal serum was replaced by 5 mg/mL of bovine serum albumin (BSA) comprised the study group. Seventy-one patients with tubal infertility served as controls. In 50 of these, medium was supplemented with 7.5% maternal serum, and 21 were assigned to BSA substitution. RESULTS: Percentage of fertilization in the study group was significantly lower (41 +/- 31; mean +/- SD) than that of controls with maternal serum (77 +/- 15) and BSA (76 +/- 22). Early embryonic quality, as assessed by percentage of cleavage and morphological grading, was found to be inferior in patients with antisperm antibodies. The percentage of advanced embryos (greater than or equal to 4 blastomeres) at the time of transfer was 42 +/- 39 in the study group, compared with 65 +/- 23 and 75 +/- 35 for maternal serum and BSA controls, respectively. Percentage of morphologically favorable embryos (grades 1 and 2 in a 1 to 5 grading system) was 49 +/- 31 in the study group, compared with 78 +/- 35 and 74 +/- 23 for the controls. Percentage of clinical pregnancy was somewhat lower in the study group (12.5%) than in controls with either maternal serum (18%) or BSA (19%). CONCLUSIONS: Antisperm antibodies may have an adverse effect on fertilization and early embryonic development. Female immunological infertility may not be completely alleviated by IVF-ET.

Adult↗

A novel subfamily of zinc finger genes involved in embryonic development.

C2H2 zinc finger proteins make up one of the largest protein families in eukaryotic organisms. Recent study in several different systems has identified a set of novel zinc finger proteins that appear to form a distinct subfamily that we have named the NET family. Members of the NET family (Noc, Nlz, Elbow, and Tlp-1) share two protein motifs--a buttonhead box and an Sp motif--with zinc finger proteins from the Sp family. However, the NET family is uniquely characterized by a single atypical C2H2 zinc finger, in contrast to the Sp family that contains three tandem C2H2 fingers. Here, we review current information about the biochemical function and in vivo role for members of this subfamily. In general, NET family proteins are required during embryonic development. They appear to act by regulating transcription, most likely as repressors, although they are unlikely to bind DNA directly. In the future, it will be important to directly test if NET family proteins control transcription of specific target genes, perhaps via interactions with DNA-binding transcription factors, as well as to further explore their function in vivo.

Animals↗

The effects of simulated microgravity on avian embryonic development.

Based on the few reports available, microgravity (MG) can have adverse effects on the early development of vascularised extra-embryonic membranes in avian eggs. Whether gravity or oxygen availability is the stimulus for development of the blood vessels in the chorioallantoic membranes (CAM) remains unclear. Under gravity the blastoderm forms on top of the yolk sac, closest to the oxygen rich region beneath the shell membranes, and from there the CAM buds from an abdominal extension subsequently to form a close contact with shell membranes. Then as the embryo develops it spreads beneath the eggshell surface to maximise the surface area of the CAM vascular bed available for O2 uptake. To investigate how simulated MG influences development of the CAM and embryo we conducted experiments on chicken embryos during incubation in a 3D-clinostat (control or continuous MG treatment at 5 rpm). Further, to determine if CAM angiogenesis is directed towards regions of high O2 tension or gravity we investigated the effects of wax treatment (50% shell surface area) on development in MG. We found that clinostat MG caused embryonic failure between day 0-5 by preventing normal development of CAM-shell membrane complex. Thereafter acute MG promoted increases in CAM mass, but did not affect embryo mass. Preliminary findings suggest that combined acute MG and wax treatment did not significantly affect embryonic growth in either MG or control groups, but retarded CAM growth in control embryos only. Finally, we will present evidence to show that acute and prolonged exposure to MG does not prevent normal growth and hatching, but might have more subtle effects on hatchling physiology, including reduced heart mass.

Acrylic Resins↗

Mef2c is a direct transcriptional target of ISL1 and GATA factors in the anterior heart field during mouse embryonic development.

The vertebrate heart forms initially as a linear tube derived from a primary heart field in the lateral mesoderm. Recent studies in mouse and chick have demonstrated that the outflow tract and right ventricle originate from a separate source of mesoderm that is anterior to the primary heart field. The discovery of this anterior, or secondary, heart field has led to a greater understanding of the morphogenetic events involved in heart formation; however, many of the underlying molecular events controlling these processes remain to be determined. The MADS domain transcription factor MEF2C is required for proper formation of the cardiac outflow tract and right ventricle, suggesting a key role in anterior heart field development. Therefore, as a first step toward identifying the transcriptional pathways upstream of MEF2C, we introduced a lacZ reporter gene into a bacterial artificial chromosome (BAC) encompassing the murine Mef2c locus and used this recombinant to generate transgenic mice. This BAC transgene was sufficient to recapitulate endogenous Mef2c expression, and comparative sequence analyses revealed multiple regions of significant conservation in the noncoding regions of the BAC. We show that one of these conserved noncoding regions represents a transcriptional enhancer that is sufficient to direct expression of lacZ exclusively to the anterior heart field throughout embryonic development. This conserved enhancer contains two consensus GATA binding sites that are efficiently bound by the zinc finger transcription factor GATA4 and are completely required for enhancer function in vivo. This enhancer also contains two perfect consensus sites for the LIM-homeodomain protein ISL1. We show that these elements are specifically bound by ISL1 and are essential for enhancer function in transgenic embryos. Thus, these findings establish Mef2c as the first direct transcriptional target of ISL1 in the anterior heart field and support a model in which GATA factors and ISL1 serve as the earliest transcriptional regulators controlling outflow tract and right ventricle development.

Animals↗

Actin gene in prawn, Macrobrachium rosenbergii: characteristics and differential tissue expression during embryonic development.

An actin gene (named Mar-actin) from the commercial prawn, Macrobrachium rosenbergii, was isolated, sequenced and gene expression was characterized. The cDNA sequence was 1281 bp in length and contained 1131 bp open reading frame encoding 376 amino acids. The amino acid sequence deduced from the nucleotide sequence showed high identity (70.3% to 98.1%) with other known actins of various organisms, highest with that of the European flounder (98.1%). The 3' untranslated region (3' UTR) of the Mar-actin mRNA has a high A+U content (approximately 78%) and contains one AUUUUUA and two repeats of the AUUUA motifs, that may function in regulating mRNA decay. Northern blot analysis revealed that the Mar-actin gene was expressed predominantly in muscle tissues. Transcripts in hepatopancreas were barely detectable. Expression of the Mar-actin gene varied during embryonic development and reached the maximal level at the zoea stage. This is the first report describing the complete sequence and expression pattern of the actin gene in prawns.

Actins↗