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Influence of superovulation on endometrial and embryonic development.

The authors have studied the temporal relationship between follicular rupture and endometrial development in 13 women during a natural ovarian cycle (length 25 to 35 days), and subsequently after standard treatment with clomiphene citrate, human menopausal gonadotropin and human chorionic gonadotropin (hCG) to induce multiple folliculogenesis for oocyte recovery, in vitro fertilization, and embryo freezing (cycle length 23 to 27 days). An endometrial biopsy was taken during both cycles 1.5 to 2.0 days after the oocytes had been released or removed. The samples were examined by light and transmission electron microscopy. Samples of peripheral blood were taken at defined times for hormone analysis. After treatment 11 subjects (85%) had advanced morphological development of the endometrium (8 women by 3 to 4 days, 3 women by 1 to 2 days). The concentrations of plasma estradiol (E2) and progesterone (P) on the days of follicular rupture and endometrial biopsy were significantly raised in the treatment cycles. The concentration of total urinary estrogens on the day of hCG administration and the mean change in the concentration of plasma E2 (treatment/control) on the days of endometrial biopsy were positively correlated with the extent of endometrial advancement. In addition, the mean change in the concentration of plasma P (treatment/control) was markedly increased on the days of follicular rupture and endometrial biopsy in those subjects with an advanced endometrium. Embryonic development was not so obviously related to the extent of superovulation. Asynchronous endometrial and embryonic development may therefore contribute to the low pregnancy rate in these patients.

Adult

Embryonic development of four different subsets of cholinergic neurons in rat cervical spinal cord.

The developmental stage at which a neuron becomes committed to a neurotransmitter phenotype is an important time in its ontogenetic history. The present study examines when choline acetyltransferase (ChAT) is first detected within each of four different subsets of cholinergic neurons previously identified in the cervical enlargement of the spinal cord: namely, motor neurons, partition cells, central canal cluster cells, and dorsal horn neurons. By examining the temporal sequence of embryonic development of these cholinergic neurons, we can infer the relationships between ChAT expression and other important developmental events. ChAT was first detected reliably on embryonic day 13 (E13) by both biochemical and immunocytochemical methods, and it was localized predominantly within motor neurons. A second group of primitive-appearing ChAT-positive cells was detected adjacent to the ventricular zone on E14. These neurons seemed to disperse laterally into the intermediate zone by E15, and, on the basis of their location, were tentatively identified as partition cells. A third group of primitive ChAT-immunoreactive cells was detected on E16, both within and around the ventral half of the ventricular zone. By E17, some members of this "U"-shaped group appeared to have dispersed dorsally and laterally, probably giving rise to dorsal horn neurons as well as dorsal central canal cluster cells. Other members of this group remained near the ventral ventricular zone, most likely differentiating into ventral central canal cluster cells. Combined findings from the present study and a previous investigation of neurogenesis (Phelps et al.: J. Comp. Neurol. 273:459-472, '88), suggest that premitotic precursor cells have not yet acquired the cholinergic phenotype because ChAT is not detectable until after the onset of neuronal generation for each of the respective subsets of cholinergic neurons. However, ChAT is expressed in primitive bipolar neurons located within or adjacent to the germinal epithelium. Transitional stages of embryonic development suggest that these primitive ChAT-positive cells migrate to different locations within the intermediate zone to differentiate into the various subsets of mature cholinergic neurons. Therefore, it seems likely that spinal cholinergic neurons are committed to the cholinergic phenotype at pre- or early migratory stages of their development. Our results also hint that the subsets of cholinergic cells may follow different migration routes. For example, presumptive partition cells may use radial glial processes for guidance, whereas dorsal horn neurons may migrate along nerve fibers of the commissural pathway. Cell-cell interactions along such diverse migratory pathways could play a role in determining the different morphological, and presumably functional, phenotypes expressed by spinal cholinergic neurons.

Animals

The embryonic development of the lateral nasal wall from 8 to 24 weeks.

This histological study of 20 fetal heads aged between 8 and 24 weeks of gestation demonstrates and describes the embryonic development of the lateral wall of the nose. The three turbinates (inferior, middle, and superior) arise as soft-tissue swellings (preturbinates) by 8 weeks' gestation. A cartilage capsule surrounds the nose at 8 weeks and by 9 weeks, medially directed flanges of cartilage have invaded all three preturbinates. The uncinate process arises from the medial surface of the lateral cartilaginous capsule and is first identifiable by 10 weeks. An "air space" progressively develops from 11 to 12 weeks lateral to the cartilaginous uncinate process and from this space, the embryonic channel to the maxillary sinus develops. The embryonic woven bone of the maxilla can be identified from 9 to 10 weeks and enlarges both absolutely and relatively to the nasal cavity, so that by 13 to 14 weeks, this expanding bone forms the lateral wall of the inferior meatus as the cartilaginous nasal capsule regresses.

Embryonic and Fetal Development

Peculiarities of the embryonic development of Polypodium hydriforme Ussov (Coelenterata), a parasite of acipenserid oocytes.

"Unicellular" stages (107 specimens) and multicellular stages (64 specimens) of embryogenesis of Polypodium, found in 14 sterlet (Acipenser ruthenus L.) females, have been studied with light microscopy, cytophotometry, and autoradiography following incubation with 3H-uridine. All stages of the embryonic development occur inside host oocytes. The "unicellular" stage includes a binucleate cell with unequally sized nuclei; separation inside it of a small cell around the smaller nucleus, i.e. transformation of the single cell into a complex of 2 cells, the larger one enveloping the smaller; formation of a cavity inside the nucleus of the large (outer) cell, and migration of the small cell into it, and "cell-in-a-cell" stage, the small (generative) cell being inside the cavity formed by the nucleus of the large (trophic) cell. The latter gives rise to a hypertrophied but still unicellular envelope around the embryo, the trophamnion. The multicellular stages start with segmentation of the generative cell into blastomeres. These form a morula lying inside the cavity of the trophamnion. Gastrulation occurs by morular delamination. The inversion of the germ layers, typical of parasitic Polypodium stages, apparently arises during gastrulation. Both the generative cell ("egg") and the blastomeres are haploid, at least until the morula stage. The eggs of Polypodium are the smallest ones among coelenterates; they lack yolk and develop without fertilization. Diploidy seems to be restored during segmentation. The trophamnion cell grows, its nucleus becomes highly polypoid, and its cytoplasm accumulates mucoprotein inclusions. Both the blastomere nuclei and the trophamnion nucleus have large nucleoli and actively synthesize RNA. The stages of embryogenesis of Polypodium closely correspond to stages of the host oogenesis. The embryonic development of Polypodium lasts several years and is the slowest among coelenterates. However, it has some features typical of the class Hydrozoa.

Animals

Influence of 13-cis and all-trans retinoic acid on rat embryonic development in vitro: correlation with isomerisation and drug transfer to the embryo.

In vitro experiments using whole rat embryo cultures show that all-trans retinoic acid (all-trans RA) administered at low concentrations (30 ng/ml culture medium) is 10 times more active than 13-cis retinoic acid (13-cis RA) and 3 times more active when administered at high concentrations (1000 ng/ml culture medium). Morphological investigation of the embryos shows that both substances directly influence embryonic development in an identical manner. Isomerisation products of the administered compounds (all-trans RA from 13-cis RA and vice versa) were detected by HPLC both in the culture medium and the embryo. Correlation of embryonic retinoid concentration with the observed effects led us to suggest that the isomerisation to all-trans RA is crucial in regard to 13-cis RA-induced abnormal embryonic development. A 100% effect can be induced in vitro with very low amounts of all-trans RA (7.2 ng/g) in the embryo.

Animals

Changes in the electrical properties of chick ciliary ganglion neurones during embryonic development.

1. Whole-cell recording techniques were used to examine the expression of ionic currents in chick ciliary ganglion neurones dissociated acutely at various stages of embryonic development. Currents were also examined in dissociated cells that had been maintained in vitro for several days. 2. Voltage-activated, tetrodotoxin (TTX)-sensitive Na+ currents (INa) could be detected in all cells tested between stage 25 and stage 40 (embryonic days 4.5-14). INa increased in both amplitude and density throughout development, but no obvious changes in kinetics or sensitivity to TTX were observed. 3. High-threshold Ca2+ currents (ICa) were also detectable between stage 25 and stage 40. ICa increased in both amplitude and density throughout this time. No obvious changes in kinetics or voltage dependence were observed. 4. Delayed rectifier K+ currents (IDR) and A-currents (IA) could be detected in Ca(2+)-free salines, and distinguished on the basis of differences in kinetics, voltage dependence, and sensitivity to tetraethylammonium (TEA). IA was either absent, or present at very low densities at stages 26-30, but showed a sharp increase in density thereafter. In contrast, IDR was detectable as early as stage 25, and did not display a significant increase in density during development. 5. Ca(2+)-activated K+ currents (IK(Ca)) were either undetectable or present at very low density between stage 26 and stage 30 (embryonic days 5-9) but showed a large increase in amplitude and density thereafter. 6. Ionic currents were examined in age-matched cells dissociated acutely on embryonic day 13, or isolated on embryonic day 9 and maintained in vitro for an additional 4 days. Most of the cells maintained in culture for 4 days did not express detectable IK(Ca), and had significantly reduced IA compared to acutely isolated controls. The cultured cells expressed normal densities of IDR, ICa and INa. 7. All ionic currents increased in amplitude during normal embryonic development, and all but IDR increased in density. The largest change in density generally occurred between stages 30 and 40, during which time ciliary ganglion neurones form synapses with target tissues. 8. Isolation of ciliary neurones from the in ovo environment prevented the normal development of IA and IK(Ca), suggesting that the expression of these channels is controlled by one or more extrinsic environmental factors. In contrast, the normal expression of INa, ICa and IDR is not dependent upon extrinsic factors.

Animals

Embryonic development of rat diaphragm. An electron-microscopic study.

Ultrastructural aspects of white Wistar rats diaphragm during part of its embryonic development (from the 13th embryonic day till birth) have been studied. The dominating structures observed in the period of the thirteenth embryonic day (ED 13) are undifferentiated cells, their cytoplasm being poor in organelles but rich in ribosomes. The close examination of these cells reveals that some of them possess a kind of thin filaments near their Golgi zones. At ED 14-15 clusters of myoblasts are readily detected (their cytoplasm containing a lot of glycogen granules and myofibrils, some of them even with Z-line material); contact sites between their cell membranes appear, somewhere forming specialized junctions; in this period the myoblasts start to fuse giving rise to the primary generation of myotubes. At ED 16-17 the quantity of myofilaments and glycogen granules increases alongside with the initiation of a basal-lamina formation; occasionally some oval, undifferentiated cells very similar to those viewed at ED 13 are found. At ED 18-19 the cytoplasm of the myotubes contains a lot of myofibrils and a well-developed endoplasmic reticulum; at some places the adjacent membranes still form deep interdigitations. At the end of the prenatal myogenesis (ED 20-21) most of the muscle cells are close to their mature morphological appearance--the sarcomers are well-organized and some nuclei present a peripheral localization; nevertheless, in this period new generations of myotubes can be also distinguished.

Animals

Expression of murine Ia antigens during embryonic development.

An immunochemical analysis of the kinetics of appearance of Ia antigens during embryonic development was performed. Ia antigens first appear on the surface of embryonic cells 11 days postconception and their expression between days 11 and 16 of gestation is confined to the fetal liver. Ia antigen synthesis by fetal liver cells is detectable at day 14. Ia seems to precede Ig as a surface marker of embryonic liver cells, since Ig cannot be detected until day 16 of gestation. H-2 antigens may be immunoprecipitated from day 10 whole embryo cells. F9 primitive teratocarcinoma cells are Ia negative and H-2 negative.

Aging

'Cleavage fields': hypothesis on early embryonic development.

The hypothesis is put forward that events of the early embryonic development can be interpreted on the basis of a radial distribution of cytoplasmic components, i.e. of a 'cleavage field', progressively established during the growth of the oocyte. The orientations of the cleavage spindles and the corresponding furrows' positions are assumed to be correlated to the field's temporal evolution which, in turn, is determined by flows of cytoplasmic components originated by the changes in the membrane shape. From this viewpoint, a simple explanation of egg regulation is proposed, and the particular case of the sea urchin embryo is briefly discussed.

Animals

[Effect of glycerol formal on the embryonic development of the rat].

In order to evaluate the effect on embryonic development of a solvent commonly used in pharmacological investigations, glycerol formal was administered to pregnant rats from days 6 through 15 of gestation at the daily doses of 0.25, 0.50 and 1.0 ml/Kg i.m. Glycerol formal did not induce systemic toxicity in the mothers, but showed an embryotoxic and teratogenic activity on the products of conception.

Animals

Effect of the number of inseminated spermatozoa on subsequent human and mouse embryonic development in vitro.

It has been shown, in both human and mouse in-vitro fertilization (IVF), that an excess number of spermatozoa in the insemination medium leads to reduced fertilization rates. In this study, we evaluated human embryonic development after dividing the oocytes of each of 62 IVF attempts into two groups on the basis of insemination with two widely used concentrations (50,000 and 100,000 spermatozoa/ml). The embryonic growth was retarded in the group inseminated with 100,000 spermatozoa/ml: significantly fewer fast developing embryos (4-cell and 5- to 8-cell stages) were found (53.4% in the 100,000/ml group and 65.5% in the 50,000 group; P less than 0.05). In two experimental series, mouse embryonic development was evaluated in the presence of 0, 50,000, 100,000 and 500,000 spermatozoa per ml. In the first series, the spermatozoa were present during 5-20 h after insemination, while in the second series, the spermatozoa were present during the whole culture period of 120 h. The development of mouse embryos was impaired when 500,000/ml spermatozoa were present during the whole culture period. In contrast with human IVF results, the presence of up to 500,000 spermatozoa during the first 20 h after insemination did not have any significant detrimental effect on blastocyst formation in the mouse.

Animals

Changes in DNA methylation during mouse embryonic development in relation to X-chromosome activity and imprinting.

Changing DNA methylation patterns during embryonic development are discussed in relation to differential gene expression, changes in X-chromosome activity and genomic imprinting. Sperm DNA is more methylated than oocyte DNA, both overall and for specific sequences. The methylation difference between the gametes could be one of the mechanisms (along with chromatin structure) regulating initial differences in expression of parental alleles in early development. There is a loss of methylation during development from the morula to the blastocyst and a marked decrease in methylase activity. De novo methylation becomes apparent around the time of implantation and occurs to a lesser extent in extra-embryonic tissue DNA. In embryonic DNA, de novo methylation begins at the time of random X-chromosome inactivation but it continues to occur after X-chromosome inactivation and may be a mechanism that irreversibly fixes specific patterns of gene expression and X-chromosome inactivity in the female. The germ line is probably delineated before extensive de novo methylation and hence escapes this process. The marked undermethylation of the germ line DNA may be a prerequisite for X-chromosome reactivation. The process underlying reactivation and removal of parent-specific patterns of gene expression may be changes in chromatin configuration associated with meiosis and a general reprogramming of the germ line to developmental totipotency.

Animals

The effect of copper on the embryonic development and hatching of Sepia officinalis L.

The influence of copper on embryonic development and hatching of Sepia offinalis was investigated. Copper exerts a profound effect on both hatching stage and time-to-hatching. At high copper concentrations (50-200 ppb Cu2+), the embryos hatch earlier than the controls but have a lower survival potential. No external morphological malformations were found. Whereas copper does not accumulate in the embryo or in the vitellus, it is absorbed by the envelope and/or the chorion.

Animals

Post-embryonic development of rectifying electrical synapses in the crayfish: ultrastructure.

The post-embryonic development of the rectifying Giant Fibre-Motor Giant (GF-MoG) synapse and the Giant Fibre-Segmental Giant (GF-SG) synapse has been investigated using electron-microscopy. In adults, the MoG and SG neurons make contact with the GFs by sending narrow 'finger-like' processes through the glial and connective tissue sheath surrounding each GF. The junctional region is characterized by closely apposed membranes (approximately 4 nm separation) traversed by regularly spaced connections, and large (60-80 nm) spherical vesicles in the presynaptic fibre. In newly hatched crayfish junctional contact is made over extensive areas of flat membrane apposition, due to the absence of a thick connective sheath around the giant fibres. Initially the junctional region is dominated by contacts which are morphologically indistinguishable from chemical synapses, i.e. 1. The apposed membranes are separated by a cleft of approximately 20-30 nm (an order of magnitude larger than the cleft distance at electrotonic synapses). 2. There is pre- and post-synaptic thickening of the junctional membranes with a dense cytoplasmic material. 3. Small (25-40 nm) pleomorphic vesicles are found on the presynaptic side of the junction, commonly in association with a dense presynaptic bar. Regions of junctional contact displaying the adult electronic-type morphology first appear at approximately one week post-hatching. At this age they are limited in distribution and occupy a central position in the area of contact surrounded by a broad 'chemical-like' annulus. During subsequent development these sites with electrotonic-type morphology grow in relative size, so that the 'chemical-like' sites become compressed towards the edges of the regions of contact. The adult type of morphology, in which the 'chemical-like' regions are vestigial, is achieved approximately two months after hatching.

Animals

Sialic acid at the surface of myocardial cells during embryonic development.

We tested the hypothesis that the reduction of automaticity during the embryonic development of chick ventricular myocytes is correlated with the number of sialic acid residues at the cell surface. The major findings were twofold. First, the sialic acid content of ventricular tissue fragments declined during the period between 4 and 17 days of development; however, when a 26% reduction of cell surface area was taken into account, the surface density of sialic acid at 7 and 17 days was not significantly different. Second, the sialic acid content of ventricular cell aggregates (after 3 days in gyratory culture) increased during the same two-week period. On the surface of these cells, the density was significantly greater at 17 days than at 7 days, even after a 17% increase in cell surface area had been taken into account. When the developmental increase in sialic acid content was compared with a concomitant decline in aggregate beat rates, we calculated a correlation coefficient of 0.85. Thus, while there could be some relationship between aggregate automaticity and sialic acid content, there appears to be no such correlation for fragments of chick ventricle.

Animals

The role of transposable elements-endogenous retroviruses in embryonic development and regeneration.

Endogenous retroviruses (ERVs) are dynamically regulated across the lifespan and can function as context-dependent components of host gene-regulatory networks. During embryonic development, selected ERV-derived elements are co-opted to support zygotic genome activation, lineage specification, and placental development. In adult tissues, ERV-derived sequences can contribute to tissue and immune homeostasis, whereas potentially disruptive ERV activity is constrained by epigenetic mechanisms. During regeneration and somatic cell reprogramming, ERV and broader transposable-element programs undergo transient, locus-specific remodeling. In aging, the weakening of epigenetic and nuclear restraint can promote aberrant ERV derepression, inflammation, and functional decline. This review summarizes the diverse roles of ERVs across these contexts and discusses the challenges of defining locus-specific functions, resolving repetitive sequences, and developing safe ERV-targeted interventions.

Endogenous Retroviruses

Inhibition of polyisoprenoid and glycoprotein biosynthesis causes abnormal embryonic development.

Compactin, a potent inhibitor of polyisoprenoid biosynthesis, induces abnormal gastrulation during sea urchin development at concentrations that have no effect on earlier embryonic development or on macromolecular synthesis. Three lines of evidence suggest that the developmental lesion caused by compactin results from inhibition of dolichol biosynthesis and a concomitant inhibition in the biosynthesis of the oligosaccharide chains of N-linked glycoproteins. (i) Embryos cultured in the presence of compactin gastrulate normally when supplemented with dolichol alone, whereas supplementation with cholesterol or coenzyme Q or both does not prevent the compactin-induced developmental lesion. (ii) Exogenously supplemented [3H]dolichol is incorporated into a compound with the chromatographic properties of oligosaccharide-pyrophosphoryldolichol. (iii) Embryos cultured in the presence of compactin exhibit a decreased capacity to synthesize mannose-labeled glycolipids and N-linked glycoproteins. This decrease in synthesis is abolished if the embryos are cultured in the presence of dolichol along with compactin.

Animals