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At least 325 records · Page 18Linked to original sources

PubMed2005

Assessment of ploidy stability of the somatic embryogenesis process in Quercus suber L. using flow cytometry.

Flow cytometry analyses were used to verify the ploidy stability of Quercus suber L. somatic embryogenesis process. Leaf explants of two adult cork oak trees (QsG0 and QsG5) of the North of Portugal were inoculated on MS medium with 2,4-D and zeatin. After 3 months, calluses with embryogenic structures were isolated and transferred to fresh MS medium without growth regulators and somatic embryo evolution was followed. Morphologically normal somatic embryos (with two cotyledons) and abnormal somatic embryos (with one or three cotyledons) were used in this assay. Flow cytometry combined with propidium iodide staining was employed to estimate DNA ploidy levels and nuclear DNA content of somatic embryos and leaves from mother plants. No significant differences (P< or =0.05) were detected among embryos, and between the embryos and the mother plants. Also, after conversion of these embryos, no significant morphological differences were observed among the somatic embryo-derived plants. These results and further studies using converted plantlet leaves and embryogenic callus tissue indicate that embryo cultures and converted plantlets were stable with regard to ploidy level. As no major somaclonal variation was detected our primary goal of "true-to-type" propagation of cork oak using somatic embryogenesis was assured at this level. The estimation of the 2C nuclear DNA content for this species is similar to the previously obtained value.

Flow Cytometry↗

PubMed1998

Extracellular beta-1,3-glucanases are induced during early somatic embryogenesis in Cichorium.

In leaf tissues of the Cichorium hybrid clone '474' (C. intybus L. var. sativum x C. endivia L. var. latifolia), the acquisition and expression of embryogenic competence was characterised by the appearance of 15 polypeptides (Boyer et al., 1993, Plant Sci 93: 41-53). The 38-kDa proteins were found to be abundantly present in conditioned embryogenic medium after the first division of the induced cells. These proteins seemed to be glycosylated as indicated by general carbohydrate detection methods. Internal amino-acid sequences obtained after microsequencing tryptic peptides appeared to be 36-57% homologous with plant beta-1,3-endoglucanases. In addition, these 38-kDa proteins were recognised by antibodies raised against the pathogenesis-related tobacco glucanase PR2a and their beta-1,3-glucanase activity was demonstrated by direct detection in polyacrylamide gels after electrophoresis. These results strongly suggested that the 38-kDa somatic-embryogenesis-related (SER) polypeptides are beta-1,3-glucanases. Moreover, the level of glucanase activity was nearly three times higher in the medium of the embryogenic '474' line than in the medium of a non-embryogenic line. The possible involvement of the extracellular 38-kDa proteins in callose degradation during somatic embryogenesis is discussed.

Amino Acid Sequence↗

PubMed2002

Comparative and experimental embryogenesis of Plectidae (Nematoda).

Comparative analysis of early embryogenesis indicates that considerable differences exist among nematode species. To better understand to what extent the well-studied development of Caenorhabditis elegans is representative for nematodes in general, we extended our earlier studies to other families of this phylum. Here we report our findings on seven species of Plectidae. We found that Plectidae embryos share a number of developmental similarities with one branch of nematodes (Secernentea), including C. elegans, but not with the other branch (Adenophorea), and thus support conclusions concerning their phylogenetic position drawn from molecular data. However, Plectidae also show developmental differences to other Secernentea, suggesting an early separation from them. Prominent characteristics of Plectidae are (1) strict left-right divisions of somatic founder cells generating a prominent early bilateral symmetry and (2) a very early start of gastrulation with immigration of a single gut precursor cell. To determine whether gastrulation with two gut precursors is crucial for C. elegans embryos, we induced it to gastrulate with a single blastomere like in Plectidae. As this alteration is compatible with an essentially normal subsequent embryogenesis, cleavage of the gut precursor before gastrulation is obviously not required. As major differences exist among nematodes concerning the potential to compensate for eliminated early blastomeres, we tested this feature in one Plectus species. We found that Plectus does not replace a lost cell but behaves like C. elegansin this respect, in contrast to our previous findings in Acrobeloides nanus, another member of the Secernentea.

Animals↗

PubMed1998

Distribution of murine mannose receptor expression from early embryogenesis through to adulthood.

The mannose receptor is a 175-kDa transmembrane glycoprotein that appears to be expressed on the surface of terminally differentiated macrophages and Langerhans cells. The ectodomain of the mannose receptor has eight carbohydrate recognition domains. The receptor recognizes the patterns of sugars that adorn a wide array of bacteria, parasites, yeast, fungi, and mannosylated ligands. Clearance studies in whole animals have localized radiolabeled ligands, such as mannosylated bovine serum albumen, not only to macrophages, but also to liver sinusoidal endothelial cells. Hitherto, there has been no comprehensive analysis of expression of the mannose receptor in embryonic and adult mouse tissues. In this study, we have undertaken a systematic survey of the expression of the mannose receptor from early embryogenesis through to adulthood. The mannose receptor is expressed on tissue macrophages throughout the adult mouse as expected. However, the mannose receptor is first observed on embryonic day 9 on cells that line blood island vessel walls in the yolk sac. The mannose receptor is localized on sinusoidal endothelial cells in embryonic liver by embryonic day 11 and in bone marrow at embryonic day 17. This pattern persists in these organs throughout embryogenesis into adulthood when sinusoidal endothelial cells of lymph nodes also express the mannose receptor. The receptor is also found on lymphatic endothelial cells of small intestine. In contrast, sinusoids of spleen and thymus do not express mannose receptor antigen. This study demonstrates that the mannose receptor is expressed on tissue macrophages and on subsets of vascular and lymphatic endothelial cells. Thus, the mannose receptor maybe a marker of the so-called reticuloendothelial system.

Aging↗

PubMed1999

Embryogenesis of pancreaticobiliary maljunction inferred from development of duodenal atresia.

The embryogenesis of pancreaticobiliary maljunction is inferred from the embryogenesis of duodenal atresia. The epithelial cells of the duodenum begin to proliferate and completely plug the lumen, but a process of vacuolation recanalizes the duodenum. Recanalization of the common duct frequently appears with two lumina and openings into the duodenum with two orifices. These two major canals create a narrow segment of the duodenum and this narrow zone is the area in the duodenum that is most prone to faulty recanalization and atresia formation. A bifid biliary system inserts at blind upper and lower pouches of the duodenum, and the common bile duct inserts in a Y fashion. The common bile duct inserts at the stenotic segment, and the end result is a T-shaped formation in patients with duodenal stenosis. During the development of the bile duct, abnormal fusion may occur between the bile duct and branches of the right ventral pancreatic duct. The site in the bile duct where a branch of the pancreatic duct joins is likely to develop atresia due to disturbance of the recanalization process, as seen in duodenal atresia. Severe impairment of vacuolation causes divided atretic bile duct at the site where the pancreatic duct inserts in a Y-fashion into the upper and lower bile duct. Moderate impairment of vacuolation causes a stenosis at the site where the pancreatic duct inserts in a T-shape, with a moderate dilatation of the bile duct.

Choledochal Cyst↗

PubMed2002

Gene expression of insulin-like growth factor-I receptor and p53 suppressor during zebrafish (Danio rerio) embryogenesis.

In vertebrates insulin-like growth factors (IGFs) regulate important cellular activities involving proliferation, differentiation, and antiapoptosis and their biological activities are mediated through the insulin-like growth factor-I receptor (IGF-IR). To understand the functions of IGF-IR in zebrafish embryogenesis, the polymerase chain reaction (PCR) cloning technique was applied to isolate the IGF-IR gene. A 5'-truncated 3285-nucleotide zebrafish IGF-IR sequence was assembled from 3 overlapping clones. This contained a partial coding region of 1550 nucleotides and a 1735-nucleotide 3' untranslated region. The deduced 515 amino acid residues included the conserved kinase domain and shared 60.9%, 61.1%, and 59.9% homology to human, mouse, and frog, respectively. To understand the relationship of IGF-IR with p53 suppressor gene during embryogenesis, expression of both genes was analyzed in parallel by semicompetitive reverse transcriptase PCR and whole-mount in situ hybridization. This analysis indicated that messenger RNA of both genes was of maternal origin, but the p53 suppressor mRNA was relatively more abundant than the IGF-IR message in most of the developmental stages, except possibly at 28 hours postfertilization. At this stage the IGF-I receptor message was highly expressed and visible in whole internal organ regions by whole-mount in situ hybridization, while p53 message was concentrated in the head portion and barely detectable in the trunk portion. The results suggest that IGF-IR and p53 mRNA are expressed at different places and different times. However, the temporal and spatial relationship of IGF-IR and its relationship to p53 suppressor protein during developmental processes remain unknown.

Journal Article↗

PubMed2006

Embryogenesis and seed development in Sinomanglietia glauca (Magnoliaceae).

The development of the floral bud, especially the ovule and seed coat, of Sinomanglietia glauca was observed. Floral buds were covered by eight to nine hypsophyll pieces. The hypsophyll nearest the tepal was closed completely and characterized by two arrays of densely stained cells with dense cytoplasm, which split longitudinally at flowering. The perianth consisted of 16 tepals arranged in three whorls. The gynoecium was composed of numerous apocarpous carpels; the ovule was anatropous with two integuments. Embryogenesis was of the Polygonum type, and the endosperm was nuclear. The inner integument degenerated during seed development. The seed of S. glauca had an endotestal seed coat comprised of a sclerotic layer derived from the inner adaxial epidermis of the outer integument and a sarcotesta derived mainly from the middle cells between the inner and outer epidermis of the outer integument. The embryo developed normally, so embryogenesis is not the cause of difficult regeneration.

Flowers↗

PubMed2005

Toxicity of organic compounds to marine invertebrate embryos and larvae: a comparison between the sea urchin embryogenesis bioassay and alternative test species.

This study investigated the toxic effects of the insecticides lindane and chlorpyrifos, the herbicide diuron, the organometallic antifoulant tributyltin (TBT), and the surfactant sodium dodecyl sulfate (SDS) on the early life stages of Paracentrotus lividus (Echinodermata, Euechinoidea), Ciona intestinalis (Chordata, Ascidiacea), Maja squinado and Palaemon serratus (Arthropoda, Crustacea) in laboratory acute toxicity tests. The assays studied embryogenesis success from fertilized egg to normal larvae in P. lividus (48 h incubation at 20 degrees C) and C. intestinalis (24 h incubation at 20 degrees C), and larval mortality at 24 and 48 h in M. squinado and P. serratus. For P. lividus, the median effective concentrations (EC50) reducing percentages of normal larvae by 50% were: 350 microg l(-1) for chlorpyrifos, 5500 microg l(-1) for diuron, 4277 microg l(-1) for SDS, and 0.309 microg l(-1) for TBT. For C. intestinalis, the EC50 values affecting embryogenesis success were 5666 microg l(-1) for chlorpyrifos, 24,397 microg (l-1) for diuron, 4412 microg l(-1) for lindane, 5145 microg I(-1) for SDS, and 7.1 microg l(-1) for TBT. The median lethal concentrations (LC50) for M. squinado larval survival were 0.84 microg l(-1) (24 h) and 0.79 microg l(-1) (48 h) for chlorpyrifos, 2.23 microg(l(-1) (24 h) and 2.18 microg l(-1) (48 h) for lindane, and 687 microg l(-1) (48 h) for SDS. For P. serratus the LC50 values obtained were 0.35 microg l(-1) (24 h) and 0.22 microg l(-1) (48 h) for chlorpyrifos, 3011 microg l(-1) (24 h) and 3044 microg l(-1) (48 h) for diuron, 5.20 microg l(-1) (24 h) and 5.59 microg l(-1) (48 h) for lindane, and 22.30 microg l(-1) (24 h) and 17.52 microg l(-1) (48 h) for TBT. Decapod larvae, as expected, were markedly more sensitive to the insecticides than sea urchins and ascidians, and SDS was the least toxic compound tested for these organisms. Lowest observed effect concentrations (LOEC) of TBT for sea urchin and ascidian embryos, chlorpyrifos and lindane for crustacean larvae, and SDS, were similar to those found in many coastal areas indicating that there would be a risk to invertebrate embryos and larvae from exposure in the field to these pollutants.

Animals↗

PubMed2001

Growth hormone (GH) action in early embryogenesis: expression of a GH-response gene in sites of GH production and action.

Growth hormone (GH) may act as a local growth factor in early embryonic development, since GH- and GH-receptor (GHR) immunoreactivity is present in all tissues and most cells of embryonic chicks during organogenesis. However, as GHR-immunoreactivity could, alternatively, reflect the presence of GH-binding proteins (GHBPs) rather than authentic receptors linked to signal transduction mechanisms, GHR immunoreactivity may not be indicative of GH target sites. The possibility that GH may act as an autocrine or paracrine factor during embryogenesis was therefore assessed in the present study by determining the presence and cellular localization of mRNA for a GH-responsive gene. The mechanism of GH action involves the induction of a number of specific GH-response genes. In chickens a novel GH-responsive gene (GHRG-1) has been identified as a marker of GH action. In situ hybridization, using a 860 bp probe for GHRG-1 mRNA, demonstrated widespread expression of the GHRG-1 gene in embryonic tissues known to contain GH- and GHR-immunoreactivity (e.g. in the spinal cord, skin, heart, liver, muscle, bone and lung). GHRG-1 mRNA was not, however, present in all cells of each tissue. It was, furthermore, not present in subepithelial cells of the esophagus and bronchus and was lacking in many spinal cord ependyma, which are also known to lack GH immunoreactivity. These results therefore support the possibility that GH acts as an autocrine/paracrine factor during early chick embryogenesis, which was hitherto thought to be a "growth-without-GH" syndrome.

Animals↗

PubMed1983

Onset of 5 S RNA gene regulation during Xenopus embryogenesis.

The transcription of 5 S RNA genes during oogenesis results in the storage of sufficient 5 S RNA in ribosomes to support subsequent embryogenesis. Xenopus oocytes of all stages synthesize oocyte-type 5 S RNA. A generalized repression of transcription occurs at meiosis and is maintained throughout early cleavage. The onset of 5 S RNA synthesis is detected at approximately the 4000-cell blastula stage (stage 9), concomitant with de novo synthesis of other species of RNA. At this developmental stage the level of 5 S RNA synthesis is low relative to the synthesis of tRNA and small nuclear RNAs. Analysis of this newly synthesized 5 S RNA reveals it to be a nearly equal mixture of oocyte and somatic 5 S RNA derived from both maternal and paternal genes. Given the 50:1 ratio of oocyte to somatic 5 S RNA genes in X. laevis, these results indicate that the majority of the oocyte 5 S RNA genes are inactivated at this time. This reflects differential transcription of the two families of 5 S RNA genes rather than post-transcriptional stability as demonstrated by the ability of a chromatin template isolated from stage 9 embryos to direct the same ratio of oocyte to somatic 5 S RNA synthesis in vitro as that observed in vivo. By completion of gastrulation, 5 S RNA synthesized in vivo and directed from chromatin in vitro is at least 90% somatic 5 S RNA. These results are consistent with a model in which the decrease in concentration of the 5 S-specific transcription factor relative to the number of 5 S RNA genes during embryogenesis contributes to the inactivation of the oocyte 5 S RNA genes.

Age Factors↗

PubMed1987

Coordinate accumulation of homeologous transcripts of seven cotton Lea gene families during embryogenesis and germination.

One of two related patterns of total transcript accumulation are seen during embryogenesis for 18 cotton Lea (Late embryogenesis-abundant) gene families in the allotetraploid cotton Gossypium hirsutum L. cv Coker 201. Coordinate accumulation in each class is complex, suggesting that Lea mRNA abundance is regulated by several events. Each of the Lea gene families probably contains two active homeologous genes (alloalleles), one in each of cotton's two subgenomes. It is of interest whether both transcripts of a Lea family are regulated the same or whether the complexity of total transcript accumulation is due to different regulation of the two transcripts. The two polypeptides encoded by the homeologous transcripts of 7 Lea families can be distinguished on two-dimensional gels. The majority of Lea transcripts in total RNAs is shown to be functional in vitro throughout development; thus in vitro translation should faithfully measure their relative abundance. The ratio of the two transcripts of each was followed during embryo maturation when Lea transcript concentration increases an average of 70-fold and during the first 12 hr of germination when Lea transcripts decline in concentration an average of 50-fold. For 6 of the Lea families, the relative level of the two transcripts is invariant throughout this period. The two constituent transcripts of the seventh, Lea9, change 5-fold in relative concentration during late maturation and 3-fold during germination. Both transcripts still follow the same temporal pattern of accumulation; only their rates of change are somewhat different. These Lea families are a random sample of the 18 described. If multiple events affect the mRNA abundance of each Lea family, such events then each affect their individual transcripts in a similar way.

Alleles↗

PubMed1989

Protein synthesis and morphogenesis are not tightly linked during embryogenesis in Fucus.

Fertilized eggs of the brown alga Fucus have long been used as model organisms for investigating the early events in the establishment of polarity and subsequent embryogenesis since large numbers of zygotes can easily be obtained. We have analyzed protein synthesis in eggs and embryos during the first day of development using two-dimensional gels and found that synthesis of 12 of the 60 most prominent proteins changed either qualitatively or quantitatively. Actin and beta-tubulin were identified by immunoblotting; synthesis of these cytoskeletal proteins was initiated at different times during the first 12 hr of development. Unique, reproducible patterns of protein synthesis observed during development in the light permitted accurate staging of developing embryos. Inhibitors such as cytochalasin and sucrose, however, blocked morphogenesis without affecting protein synthesis, and, conversely, growth in the dark delayed protein synthesis but had very little effect on the timing of morphogenesis. The data are consistent with morphogenesis and protein synthesis being relatively independent during early embryogenesis. Actinomycin D added soon after fertilization had no effect on protein synthesis 1 day later, indicating that the proteins analyzed were encoded by maternal mRNA stored in the egg.

Cell Division↗

PubMed1989

A sperm-supplied product essential for initiation of normal embryogenesis in Caenorhabditis elegans is encoded by the paternal-effect embryonic-lethal gene, spe-11.

Loss-of-function mutations in the spe-11 gene in Caenorhabditis elegans result in a paternal-effect embryonic-lethal phenotype: fertilization of wild-type oocytes by sperm from homozygous spe-11 mutant males leads to abnormal zygotic development, whereas oocytes from homozygous spe-11 hermaphrodites when fertilized by wild-type sperm develop normally. Embryos fertilized by sperm from homozygous spe-11 worms fail to complete meiosis and show defects in eggshell formation, mitotic spindle orientation, and cytokinesis. Genetic analysis suggests that the spe-11 gene is expressed before the completion of spermatogenesis and that the wild-type locus encodes a product that is present in sperm and participates, directly or indirectly, in initiating the correct program of early events in C. elegans embryos. Such an ontogenetic role of the spe-11+ gene product in early embryogenesis distinguishes spe-11 mutations from the two paternal-effect mutations identified in Drosophila, ms(3)K81 and pal, which primarily affect chromosome behavior. Analysis of spe-11 provides the first step toward genetic dissection of the functions of the sperm in early embryogenesis in C. elegans.

Animals↗

PubMed1989

Dorsal expression of the Drosophila z600 gene during early embryogenesis.

The Drosophila z600 gene is a member of an overlapping gene cluster located in the 71CD interval of chromosome 3. We have used several approaches to study the spatial and temporal expression of z600 during embryogenesis. Northern analysis shows that z600 is zygotically expressed, with gene transcripts accumulating transiently during early embryo-genesis. The localization of z600 transcripts in embryo sections by in situ hybridization reveals a dynamic pattern of RNA distribution. z600 RNA is distributed throughout the embryo during the midsyncytial blastoderm stage, becomes concentrated dorsally and posteriorly during cellularization, and persists in the dorsal-most region of the embryo during gastrulation and germ band extension. z600 transcript accumulation is altered in dorsalized or ventralized mutant embryos, suggesting that z600 is directly or indirectly regulated by genes specifying the dorsal-ventral pattern in the embryo. An analysis of germ line transformants harboring a z600-lacZ gene fusion demonstrates that a 276-bp DNA region contains the sequences responsible for dorsal activation, but not ventral repression, of z600 gene expression during early embryogenesis.

Animals↗

PubMed1992

Calmodulin transcription is limited to the nervous system during Drosophila embryogenesis.

We have examined the RNA expression pattern for the Drosophila calmodulin gene during embryogenesis by in situ hybridization to transcripts in whole embryos. Our results indicate that maternally derived calmodulin mRNA is homogeneously distributed throughout the early embryo, but that these maternal transcripts are lost by maximal germ band extension. Zygotic transcription of the gene in mid- to late-stage embryos is restricted to neural cell precursors and their progeny in both the central and peripheral nervous systems. Thus, activation of calmodulin transcription during embryonic development appears to mark a commitment to a neural fate. Northern blot analysis revealed that the two transcripts from the calmodulin gene are differentially expressed during embryogenesis. Comparison of Northern blot and in situ hybridization data indicates that the longer calmodulin mRNA is a nervous tissue-specific transcript. This suggests that neural-specific regulation of polyadenylation site usage occurs. We have also examined calmodulin expression in embryos homozygous for mutations in four loci which are known to affect nervous system development: numb, the achaete-scute complex, daughterless, and mastermind. The calmodulin transcription pattern is altered in embryos mutant for each of these loci, suggesting that regulation by these genes, either directly or indirectly, is taking place.

Animals↗

PubMed1986

Transglutaminase (TG) involvement in early embryogenesis.

Transglutaminase (TG) has been examined in different stages of preimplantation mouse embryogenesis. The specific activity of this enzyme in the soluble cellular fraction increases 2-fold from 2-cell embryos to 8-cell morulae and 4-fold from 2-cell embryos to blastocyst. The same developmental profile was seen when either N,N'-dimethylcasein or endogenous substrates were used in the TG assay. Using high-speed supernatants from different stage embryos as a source of enzyme and [3H]putrescine as acyl acceptor, the major acyl donor components were tubulin and a high molecular weight (HMW) cross-linkage product, as assessed by electrophoresis and immunoblotting. When either assembled or monomeric cytoskeleton proteins were compared as substrates, microtubules were the best acyl donors. These studies indicate that TG activity is modulated during the changing demands of blastomeres for microtubule cytoskeleton in early embryogenesis.

Animals↗

PubMed1992

Xenopus laevis oocyte G alpha subunits mRNAs. Detection and quantitation during oogenesis and early embryogenesis by competitive reverse PCR.

The expression of mRNAs coding for different Xenopus laevis oocyte G alpha subunits was analyzed by the PCR technique. Using the nucleotide sequences of five previously cloned cDNAs for oocyte G alpha subunits [FEBS Lett. 244, 188-192, 1989; FEBS Lett. 268, 27-31, 1990] and the highly sensitive reverse PCR reaction we found that G alpha o, G alpha i-1, G alpha i-3 and G alpha s species are present in oocyte stage VI, G alpha o mRNA being the most abundant transcript. G alpha o mRNA was further quantitated through oogenesis, unfertilized eggs and early embryogenesis stages by a competitive PCR reaction using an 'in vitro' deleted G alpha o mRNA as the internal standard. Using this approach we found that Xenopus G alpha o mRNA levels were constant during oogenesis and unfertilized eggs at a concentration of 3.5 pg of mRNA/stage (5 x 10(5) molecules) and diminish gradually during early embryogenesis, reaching a level of 0.3 pg in the gastrula stage. These findings show that oocyte G alpha o, and perhaps the rest of the alpha subunits, are expressed as maternal mRNAs and could play an important role in signal transduction at the beginning of oocyte cell differentiation.

Animals↗

PubMed1995

The prothymosin alpha gene is specifically expressed in ectodermal and mesodermal regions during early postimplantation mouse embryogenesis.

Prothymosin alpha (ProT alpha) is a highly acidic nuclear protein, once believed to have an extracellular immunoregulatory role but more recently implicated in cell proliferation and/or differentiation. Several recent studies have revealed that ProT alpha mRNA is present during embryogenesis. However, these studies did not investigate the spatial distribution of ProT alpha mRNA in the embryo. Here we present a detailed study of the spatial distribution of ProT alpha mRNA during the early stages of postimplantation development (6.5-12.5 dpc) of the mouse. Three findings are of particular interest. First, ProT alpha mRNA levels increase during the early postimplantation stages (6.5-8.5 dpc) of mouse embryogenesis. Second, ProT alpha mRNA is not uniformly distributed in the mouse embryo, but is present in a spatially specific manner. Third, we have observed that the mouse ProT alpha gene is expressed almost exclusively in ectodermal and mesoderm-derived structures, and not in cells which give rise to the definitive endoderm.

Animals↗