PubMed Health⌕ Search

SEARCH · PubMed Health

Results for “Embryonic Development”

Explore indexed PubMed citations for clinical trials, systematic reviews and public health research. Read source abstracts and follow each citation to its original PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 703 records · Page 39Linked to original sources

Timing mechanisms in early embryonic development.

Embryological development takes place in four dimensions and requires the existence of time measuring processes within the embryo. Evidence is accumulating that suggests that the emergence of many events during early embryonic development is controlled by timing mechanisms or developmental clocks. The purpose of this work is to review recent studies on developmental timing with speculations about underlying possible mechanisms. It is an attractive idea that the development of an embryo is timed by a single clock set in motion at fertilization, but this seems not feasible. The clock mechanism which determines the time of initiation of cellular differentiation may be independent of that for the timing of morphogenesis. The clock mechanism for cellular differentiation may be closely associated with the cycles of DNA replication, while the clock which counts the time to onset of early morphogenetic events is found in the cytoplasm. These ideas can provide a framework which may help to organize existing observations and to stimulate new experimental approaches to the problem.

Acetylcholinesterase↗

Gamma tropomyosin gene products are required for embryonic development.

The actin filament system is essential for many cellular functions, including shape, motility, cytokinesis, intracellular trafficking, and tissue organization. Tropomyosins (Tms) are rod-like components of most actin filaments that differentially affect their stability and flexibility. The Tm gene family consists of four genes, alphaTm, betaTm, gammaTm (Tm5 NM, where "NM" indicates "nonmuscle"), and deltaTm (Tm4). Multiple isoforms of the Tm family are generated by alternative splicing of three of these genes, and their expression is highly regulated. Extensive spatial and temporal sorting of Tm isoforms into different cellular compartments has been shown to occur in several cell types. We have addressed the function of the low-molecular-weight Tms encoded by the gammaTm gene by eliminating the corresponding amino-terminal coding sequences from this gene. Heterozygous mice were generated, and subsequent intercrossing of the F1 pups did not result in any viable homozygous knockouts. Genotype analysis of day 2.5 morulae also failed to detect any homozygous knockouts. We have failed in our attempts to delete the second allele and generate in vitro double-knockout cells, although 51 clones displayed homologous recombination back into the originally targeted locus. We therefore conclude that low-molecular-weight products from the gammaTm gene are essential for both embryonic development and cell survival.

Animals↗

Fatty acid synthesis is essential in embryonic development: fatty acid synthase null mutants and most of the heterozygotes die in utero.

In animals, including humans, the source of long-chain saturated fatty acids is de novo synthesis, which is mediated by fatty acid synthase (FAS), ingested food, or both. To understand the importance of de novo fatty acid synthesis, we generated FAS knockout mice. The heterozygous FAS mutants (Fasn+/-) are ostensibly normal. In Fasn+/- mice the levels of FAS mRNA and the FAS activity are approximately 50% and 35% lower, respectively, than those of WT mice; hence, FAS levels are affected by gene dosage. When the Fasn+/- mutant mice were interbred, Fasn-/- mice were not produced; thus, FAS is essential during embryonic development. Furthermore, the number of Fasn+/- progeny obtained was 70% less than predicted by Mendelian inheritance, indicating partial haploid insufficiency. Even when one of the parents was WT, the estimated loss of heterozygous progeny was 60%. This loss of Fasn+/- pups appeared to be strain-specific and became more pronounced as the heterozygous females produced more litters. Most of the Fasn-/- mutant embryos died before implantation and the Fasn+/- embryos died at various stages of their development. Feeding the breeders a diet rich in saturated fatty acids did not prevent the loss of homoor heterozygotes. These observations are very important in considering teratogenic consequences of drugs aimed at inhibiting FAS activity, to reduce either obesity or the growth of cancerous tissues.

Animals↗

Effects of estrogens during embryonal development on crowing in the domestic fowl.

In the domestic fowl, crowing is typically a male-specific vocal behavior while the females normally do not crow. These sex differences in vocalization may result from organizational actions of estrogens during specific periods of embryonic development. To further investigate the role of estrogens in differentiation of crowing and development of the acoustic characteristics of crow calls, male domestic fowls were treated on Incubation Day 8 with estradiol benzoate (EB) or either oil or saline vehicles. On the same incubation day, the female fowls were treated with an aromatase inhibitor, fadrozole, or saline vehicle. An adulthood vocalization of cocks and hens was recorded during corresponding tests of sexual behavior. The exposure to EB or fadrozole had no effect on sexual differentiation of the gonads and all fadrozole-treated hens laid eggs at a rate similar to the control hens that received saline. While the levels of plasma testosterone at adulthood did not differ in treated and untreated cocks, the incidence of crowing rate was significantly lower in cocks that were exposed to estradiol. Acoustic analysis revealed a considerable reduction in duration and acoustic energy of calls while the main frequency characteristics were not changed. Four out of the seven tested fadrozole-treated hens demonstrated regularly crow-like vocalization with shorter duration and lower energy of calls in comparison to crows of the control males. These findings point out to a role for estradiol in organization of crowing behavior and a specific temporal pattern of the crowing call.

Androgens↗

Biochemical alterations in skeletal muscle induced by 2,4-dichlorophenoxyacetic butyl ester during chick embryonic development.

2,4-Dichlorophenoxyacetic butyl ester (2,4-D b.e.) (3.1 mg/egg) was applied on fertile hen eggs before starting the incubation. Chicks hatched from treated eggs showed motor dysfunctions, postural troubles and edematous muscles. The electromyography revealed muscular weakness, prolonged motor distal latency, and myotonia. The biochemical composition of leg and complexus muscles from 1-day-old chicks was determined. A significant diminution (24%) in the glycogen level of leg muscles was produced by the treatment. There was a small increase (15%) in sarcoplasmic proteins from leg muscles and an increase of a 20 kD protein in the myofibrillar proteins from complexus muscles. Even though total lipid content was not changed, 2,4-D b.e. treatment produced a diminution of sterol esters (20%) and phosphatidylcholine (11%) and an increase of phosphatidylserine (61%), triglycerides (37%) and free fatty acids (FFA) (448%) in leg muscles. Increases of phosphatidylethanolamine (16%), sterols (58%) and FFA (267%) were detected in complexus muscles. A remarkable increase (700-1500%) of unsaturated FFA, e.g. oleic, linoleic and arachidonic acids, was observed. Considering the avian embryo lipid metabolism, it is proposed that FFA and triglycerides were accumulated because they could not be metabolized in the mitochondria. Since FFA are potent cytotoxic compounds, their increase may be a key factor in the 2,4-D b.e. toxic action in muscle and other tissues during embryonic development.

2,4-Dichlorophenoxyacetic Acid↗

Embryonic development of the ureter and bladder: acquisition of smooth muscle.

To delineate the temporal and spatial acquisition of the smooth muscle of the ureter, Sprague-Dawley rat embryos and newborn pups were immunostained with alpha-smooth muscle actin (alpha-SM actin) antibody. Alpha-SM actin expression was first detected in the urinary tract at 16 days of gestation (E16) in a thin subserosal zone about the urogenital sinus. At this time, the E16 ureter is composed of a simple cuboidal epithelium which is surrounded by 1 to 2 layers of condensed alpha-SM actin negative spindle shaped cells. No immunostaining was detected along the ureter or its intrarenal branches until the 20th day of gestation (E20). Alpha-SM actin expression in the E20 ureter exhibited regional differences. The number of alpha-SM actin positive smooth muscle cells was greatest in the distal ureter, intermediate in the mid ureter, and least in the proximal ureter near the kidney. While smooth muscle formation in the bladder was subserosal, in the ureter it was subepithelial. During postnatal life, alpha-SM actin expression increased in both organs as all periepithelial spindle cells stained positive and intensified their staining. Smooth muscle differentiation of the ureter and bladder occurs later in embryonic life than other visceral and vascular organs and occurs in an ascending fashion from the bladder to the intrarenal collecting system. It is likely that the activation of visceral smooth muscle myogenesis within the urinary tract is governed by positional information specific to the embryonic development of each organ.

Actins↗

Metaxin, a gene contiguous to both thrombospondin 3 and glucocerebrosidase, is required for embryonic development in the mouse: implications for Gaucher disease.

We have identified a murine gene, metaxin, that spans the 6-kb interval separating the glucocerebrosidase gene (GC) from the thrombospondin 3 gene on chromosome 3E3-F1. Metaxin and GC are transcribed convergently; their major polyadenylylation sites are only 431 bp apart. On the other hand, metaxin and the thrombospondin 3 gene are transcribed divergently and share a common promoter sequence. The cDNA for metaxin encodes a 317-aa protein, without either a signal sequence or consensus for N-linked glycosylation. Metaxin protein is expressed ubiquitously in tissues of the young adult mouse, but no close homologues have been found in the DNA or protein data bases. A targeted mutation (A-->G in exon 9) was introduced into GC by homologous recombination in embryonic stem cells to establish a mouse model for a mild form of Gaucher disease. A phosphoglycerate kinase-neomycin gene cassette was also inserted into the 3'-flanking region of GC as a selectable marker, at a site later identified as the terminal exon of metaxin. Mice homozygous for the combined mutations die early in gestation. Since the same amino acid mutation in humans is associated with mild type 1 Gaucher disease, we suggest that metaxin protein is likely to be essential for embryonic development in mice. Clearly, the contiguous gene organization at this locus limits targeting strategies for the production of murine models of Gaucher disease.

Amino Acid Sequence↗

Changing patterns of gap junctional intercellular communication and connexin distribution in mouse epidermis and hair follicles during embryonic development.

In the mouse embryo between embryonic days 12 (E12) and 16, regular arrays of epidermal placodes on the mystacial pad develop into whisker follicles. This system was chosen for analysis of gap junctional intercellular communication during differentiation. The patterns of communication were studied by microinjection of the tracers Lucifer yellow-CH (LY-CH) and neurobiotin (NB), while immunofluorescent staining was used to study distribution of connexins 26 and 43. Extensive communication was seen between keratinocytes in developing hair pegs or, in later-stage hair follicles, in the germinative matrix. Coupling between adjacent hair pegs via interfollicular epidermis was not observed. Coupling also became restricted as follicular cells differentiated to form outer root sheath, inner root sheath, and hair shaft. Extensive gap junctional coupling is characteristic of keratinocytes that are rapidly proliferating (as in hair pegs and germinative matrix). Follicular keratinocytes commence differentiation shortly before restriction of gap junctional coupling becomes evident. Dermal mesenchymal cells undergoing different modes of differentiation also exhibit differences in gap junctional coupling, as evidenced by poor transfer of LY-CH between cells in dermal condensations of hair follicles compared with extensive transfer elsewhere in the dermis. LY-CH and NB were not transferred between epidermal or follicular epithelium and mesenchyme, arguing against a direct role for gap junctions permeable to known second messenger molecules or nucleotides in epithelial-mesenchymal interactions in this system. The distribution of connexins 26 and 43 in epidermis and hair follicles changed during differentiation but there was no correlation with changing patterns of dye transfer, indicating an unexpected degree of complexity in the relationship between gap junctional intercellular communication and connexin protein distribution during development.

Animals↗

Influence of human chorionic gonadotropin (hCG) and hCG internalization by granulosa cells on the rate of in vitro fertilization and embryonic development of human oocytes.

Follicular fluids and granulosa cells were obtained from 28 aspirated follicles of nine women undergoing laparoscopy in an in vitro fertilization program. Follicular growth was stimulated by a human menopausal gonadotropin regimen and laparoscopy was performed 32 hours after human chorionic gonadotropin (hCG) administration. Follicular fluid 17 beta-estradiol (E2) levels were higher and hCG levels were lower in follicles with oocytes that fertilized and cleaved beyond two blastomeres (greater than two-cell group) than in those with nonfertilizable oocytes (NF group) (P less than 0.05). Compared to those from the NF group, granulosa cells from the greater than two-cell group secreted less progesterone (P) in vitro and had a fourfold increase in percentage of cells with internalized hCG. These results demonstrate that the steroidogenic capacity of granulosa cells from follicles whose oocytes fertilize and undergo accelerated embryonic development in vitro differs from the capacity of granulosa cells from NF follicles. This difference may be due to their enhanced ability to bind and subsequently internalize hCG.

Chorionic Gonadotropin↗

Histomorphometric and progesterone receptor immunohistochemical analysis in the oviduct of newly hatched chicks treated with follicle-stimulating hormone during embryonic development.

In this study we evaluated the histomorphology and ultrastructure of the oviduct of newly hatched chicks, as well as the immunohistochemical expression of progesterone receptor (PR) in this tissue after follicle-stimulating hormone (FSH) treatment on days 13, 15 and 17 of embryonic development. Results indicated a marked difference in the histology of the oviduct of newly hatched chicks treated with FSH. Magnum mucosa from these animals presented a pseudostratified epithelium with evaginations from the lumen into the epithelium and from the latter into the stroma beneath where tubular glands are formed. In contrast magnum mucosa from control animals presented columnar epithelium with no evaginations. In magnum epithelium FSH also induced the formation of cilia and microvilli projections into the lumen as well as an increase in the wall and lumen areas and in the density of nuclei per unitarea. PR immunoreactivity was only observed in the oviduct of FSH treated animals. PR was located in the nucleus of epithelial luminal cells, mucosal stromal cells and smooth muscle cells. These findings suggest that FSH induces an adequate hormonal milieu for the cytodifferentiation and PR gene expression in the chick oviduct.

Animals↗

A 31P topical magnetic resonance study of embryonic development in hens' eggs.

A 31P topical magnetic resonance study of whole chicken's eggs in vivo is described. Resonances from adenosine triphosphate, phosphocreatine, inorganic phosphate and phosphorous storage protein were observed. These changed in relative intensity as the embryo developed. No ill effects due to exposure to radiofrequency radiation or magnetic fields were observed. It is concluded that topical magnetic resonance can make a useful contribution to the study of embryonic development in avian eggs.

Adenosine Triphosphate↗

Comparative effects of neutron irradiation and X irradiation on the embryonic development of the rat.

Our aim was to compare the dose-response relationship for the embryotoxic effects of 0.43 MeV neutrons with those of 240 kVp X rays after in utero exposures during early organogenesis in the rat. At 9.5 days after conception, pregnant rats were exposed to 0.025 to 0.35 Gy 0.43 MeV neutrons at a dose rate of 0.04 to 0.07 Gy/h. Comparable biological effects were produced using 0.50 to 2.05 Gy 240 kVp X rays. Neutron irradiation produced a greater proportion of offspring with very low body weight than with malformations when compared to X rays. There were no embryotoxic effects observed at neutron exposures of 0.025, 0.049, 0.079, 0.10, 0.15 and 0.20 Gy or X-ray exposures of 0.50 and 0.96 Gy. Taken together, the results suggest that the mechanisms by which neutron irradiation affects embryonic development may, in part, be both quantitatively and qualitatively different from those by which X irradiation affects development. These results support the generalization that the embryo exhibits a nonlinear response to increasing doses of ionizing radiations during the period of early organogenesis.

Abnormalities, Radiation-Induced↗

Nonhuman primate in vitro fertilization: biochemical changes associated with embryonic development.

Little research has focused on the biochemistry of primate embryos fertilized in vitro. These studies were designed to examine alterations in cellular incorporation of leucine, uptake of steroid hormones, and overall viability of such embryos. The results may be applied to certain investigations of embryonic development in humans. Squirrel monkey ova and embryos were incubated in medium supplemented with either 3H-leucine, 3H-17 beta-estradiol (E2), or 3H-progesterone (P) and processed for autoradiographic analysis or assayed for radioactivity. Cellular incorporation of 3H-leucine (as an indication of protein synthesis), declined with oocyte maturation in vitro and remained constant at in vitro fertilization (IVF). The use of the vital dyes trypan blue and fluorescein diacetate showed high correlations with the cellular incorporation of 3H-leucine by ova, therefore demonstrating viability. Uptake of E2 and P increased at IVF. The results indicate normal metabolic development of primate embryos fertilized in vitro that is comparable to preimplantation development of other mammalian species.

Animals↗

Altered levels of a 5 S gene-specific transcription factor (TFIIIA) during oogenesis and embryonic development of Xenopus laevis.

Xenopus laevis oocytes contain a 38,000-Da protein which serves both as a 5 S gene-specific transcription initiation factor (TFIIIA) and to stabilize 5 S RNA in ribonucleoprotein complexes. Using an antibody to this protein we have measured the levels of TFIIIA during oogenesis and embryonic development in X. laevis. The maximal steady state level (approximately 10(12) molecules/oocyte) is reached early in oogenesis but drops 10- to 20-fold in later stages and another 10- to 20-fold during ovulation. The reduced amount present in the unfertilized egg remains at a nearly constant level throughout early development, but with cell division the cellular concentration drops from 3 X 10(9) to about 10(4) molecules/cell. An immunoreactive protein of the same size is also found in liver tissues and in cultured kidney cells, which also contain about 10(4) molecules/cell. By both structural (CNBr peptide analysis) and functional (transcription of 5 S genes) analyses the embryonic and kidney cell 38,000-Da factors appear indistinguishable from oocyte TFIIIA. In addition a second antigenically related protein of about 40,000 Da is found in late stage embryos, liver tissues, and adult kidney cells (where it is severalfold more abundant than the 38,000-Da TFIIIA). The chromatographic fractionation and functional analysis of the kidney cell extracts has shown that fractions containing the 38,000-Da protein support 5 S RNA synthesis while fractions containing the 40,000-Da protein do not. The significance of these findings for 5 S gene regulation is discussed with respect to the dual function of TFIIIA, the presence of rate-limiting amounts of TFIIIA, and the possibility of stage-specific factors.

Animals↗

Germ cell nuclear factor is a transcriptional repressor essential for embryonic development.

GCNF is an orphan member of the nuclear receptor superfamily. The nuclear receptor superfamily is a large superfamily of transcription factors, the majority of which are designated as orphan receptors because their ligands and functions are currently unknown. GCNF (Germ Cell Nuclear Factor) is so named because of its restricted expression pattern in the adult. In the testis, GCNF is expressed only in the post meiotic round spermatids. Likewise in the ovary, GCNF's expression is restricted to the growing oocyte. To date nothing is known of GCNF's putative ligand; however, much is known about its physiological function through the use of gene targeting. Inactivation of the GCNF gene showed that it was essential for normal embryonic development. In addition to being expressed in the germ cells of the adult, it is expressed widely throughout the embryo after gastrulation. Significant strides have also been made in understanding GCNF's mechanism of action using molecular biology. The DNA binding properties of GCNF have been investigated and its response element identified. GCNF binds as a homodimer to a direct repeat element with zero nucleotides between the reiterated sequence AGGTCA. GCNF target genes have been identified that contain this DR0 element in their promoters. Such genes as Protamines 1 and 2 and Oct4 are regulated by GCNF through this element. GCNF has been shown to be a repressor of the protamine and Oct4 genes. GCNF's repression function has been shown to be mediated by interaction with the co-repressors N-CoR and SMRT in the absence of ligand. Our current efforts are to explore GCNF function in the adult germ cells using tissue specific gene targeting to specifically knock out the GCNF gene in oocytes and spermatogenic cells. In addition, efforts are being made to identify the endogenous ligand that regulates GCNF's transcriptional properties.

Animals↗

Subnuclear targeting of Runx/Cbfa/AML factors is essential for tissue-specific differentiation during embryonic development.

Runx (Cbfa/AML) transcription factors are critical for tissue-specific gene expression. A unique targeting signal in the C terminus directs Runx factors to discrete foci within the nucleus. Using Runx2/CBFA1/AML3 and its essential role in osteogenesis as a model, we investigated the fundamental importance of fidelity of subnuclear localization for tissue differentiating activity by deleting the intranuclear targeting signal via homologous recombination. Mice homozygous for the deletion (Runx2 Delta C) do not form bone due to maturational arrest of osteoblasts. Heterozygotes do not develop clavicles, but are otherwise normal. These phenotypes are indistinguishable from those of the homozygous and heterozygous null mutants, indicating that the intranuclear targeting signal is a critical determinant for function. The expressed truncated Runx2 Delta C protein enters the nucleus and retains normal DNA binding activity, but shows complete loss of intranuclear targeting. These results demonstrate that the multifunctional N-terminal region of the Runx2 protein is not sufficient for biological activity. We conclude that subnuclear localization of Runx factors in specific foci together with associated regulatory functions is essential for control of Runx-dependent genes involved in tissue differentiation during embryonic development.

Animals↗

Effect of microinjection time during postfertilization S-phase on bovine embryonic development.

Microinjection into bovine zygotes was performed to evaluate the effects of the timing of injection during the phase of DNA replication on the subsequent in vitro development of embryos and expression of injected chicken beta-actin promoter-lac Z gene construct. The period of DNA replication of bovine zygotes, determined by 3H-thymidine incorporation, begins between 12 hr and 13 hr postinsemination (hpi) of in vitro matured oocytes, reaches a maximum from 17 hpi to 19 hpi, and is complete by 21-22 hpi. Aphidicolin, an inhibitor of DNA polymerase alpha, was used to synchronize the pronuclei and the zygote population. Treatment with aphidicolin at 9-18 hpi arrested DNA replication without affecting formation of the pronuclei or embryo development. Cycloheximide, an inhibitor of protein synthesis, was used for nucleocytoplasmic resynchronization of the aphidicolin-treated zygotes. Microinjection was performed at 15 (early), 18 (mid), and 21 (late S phase) hpi. Embryonic development was affected following each of the three microinjection times. The development of zygotes injected at 18 hpi was significantly higher (P < 0.01) after 5 days of culture than those injected at 15 hpi or 21 hpi. Expression of the marker gene was observed in the higher stage of development (> 16 cells) only in the zygotes injected at 18 hpi. At the earlier stages of development, the proportions of embryos showing expression of the foreign gene were the same for all microinjection times. In aphidicolin- and cycloheximide-treated zygotes, expression of the marker gene followed the same curve as development, i.e., expression was low when injected early or late and higher (P < 0.005) when injected in the middle of zygotic S phase. The ability of the embryos to survive microinjection and to express the marker gene as a function of hpi seems to be influenced mostly in the cytoplasm processing stage rather than the pronuclei processing stage.

Animals↗

TCL1 participates in early embryonic development and is overexpressed in human seminomas.

Overexpression of the TCL1 oncogene has been shown to play a causative role in T cell leukemias of humans and mice. The characterization of Tcl1-deficient mice in these studies indicates an important developmental role for Tcl1 in early embryogenesis. In wild-type embryos, Tcl1 is abundant in the first three mitotic cycles, during which it shuttles between nuclei and the embryo cortical regions in a cell-cycle-dependent fashion. The absence of this protein in early embryogenesis results in reduced fertility of female mice. The present studies elucidate the mechanism responsible for the reduced female fertility through analysis of the oogenesis stages and early embryo development in Tcl1-deficient mice. Even though Tcl1(-/-) females display normal oogenesis and rates of oocyte maturation/ovulation and fertilization, the lack of maternally derived Tcl1 impairs the embryo's ability to undergo normal cleavage and develop to the morula stage, especially under in vitro culture conditions. Beyond this crisis point, differentiative traits of zygotic genome activation and embryo compaction can take place normally. In contrast with this unanticipated role in early embryogenesis, we observed an overexpression of TCL1 in human seminomas. This finding suggests that TCL1 dysregulation could contribute to the development of this germinal cell cancer as well as lymphoid malignancies.

Animals↗