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Maternal age effect on early human embryonic development and blastocyst formation.

In humans, age-related decline in female fertility can be explained by a reduction in quality either of the older uterus or of the embryos arising from aging oocytes. The aim of this study was to examine the latter hypothesis, using in vitro fertilization (I.V.F.) and coculture of embryos until the blastocyst stage. We determined the blastocyst formation rate ([blastocysts/embryos on day 2]* 100) and the blastocyst expansion rate ([expanded blastocysts/blastocysts]* 100) according to the patient's age the day of I.V.F. With increase in age, the number of retrieved oocytes decreased, without alteration of the cleavage rate. In patients above age 30 years, preimplantation development to blastocysts declined due to an increase in embryo arrest at the morula stage. If blastocyst stage was reached, a negative linear relationship between blastocyst expansion rate and patient age was observed. Drops in gamete production and embryo development with increasing age led to a drastic decrease in patients having at least one expanded blastocyst (< 30 years, 82%; > or = 40 years, 36%). A high delivery rate per oocyte retrieval (25.8%) was observed in patients above age 40 years after embryo transfer at the blastocyst stage. These results give a clear indication of decline in the quality of human embryos arising from aging oocytes. The origin of this alteration is discussed in terms of chromosome abnormalities, role of maternally-inherited products from the oocyte, timing of genomic activation, and temporal pattern of gene expression during initial development of the human embryo.

Adult↗

Embryonic development of the ventral body wall and its congenital malformations.

Congenital abdominal wall defects, frequently associated with other anomalies, are found in many forms. Consequently, there is still controversy in the literature concerning nomenclature, classification, and pathogenesis. Recently, we proposed a new nomenclature and classification of abdominal wall defects based on the early development of the umbilical cord and of the ventral body wall. According to this classification the complete spectrum of abdominal wall defects, including cloacal exstrophy, bladder exstrophy, and epispadias, can be subdivided into four types: primary (thoraco-)abdominoschisis, omphalocele, body wall dysplasia, and secondary (thoraco-)abdominoschisis. Each type is characterized by its specific configuration of the placenta, the membranes, the umbilical cord, and the fetus. Anomalies such as urachal remnants and omphalomesenteric duct malformations can be explained by disturbances during later stages of umbilical cord development.

Abdominal Muscles↗

Cyclic nucleotide content of ciliary and dorsal root ganglia during embryonic development in the chick.

The concentration of cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) was measured in developing chick ciliary (CG) and dorsal root ganglia (DRG) as a function of embryonic age from day 8 through day 18 using radioimmunoassays. The concentration of cAMP and cGMP increased in both ganglia from day 8 through day 14. cAMP levels were nearly two-fold higher in DRG than in CG. Normalization of the data for ciliary ganglia to the number of cells per ganglion and calculation of their molar concentrations indicates a 48% increase in cGMP and a 3.2-fold increase in cAMP during the developmental process of natural neuronal cell death.

Animals↗

FSH priming improves oocyte maturation, but priming with FSH or hCG has no effect on subsequent embryonic development in an in vitro maturation program.

AIM: To determine whether maturation and subsequent blastocyst development of in vitro matured oocytes can be improved by in vivo follicle stimulating hormone (FSH) or human chorionic gonadotrophin (hCG) priming, using a mouse model. EXPERIMENTAL DESIGN: Five groups of oocytes were used: in vivo control, in vitro matured (IVM) control, IVM after 24 h in vivo priming with FSH, IVM after 48 h in vivo priming with FSH and IVM after 16 h in vivo priming with hCG. In vitro fertilization (IVF) was performed on all groups. Oocyte maturation, fertilization, blastocyst development rates and blastocyst cell numbers were assessed for all groups. RESULTS: Significant improvement in oocyte maturation was observed in the two FSH priming groups compared with the IVM control group (P<0.005 and P<0.001, respectively). There were no significant differences in fertilization between all five groups. Blastocyst development was significantly higher in the in vivo control compared to the IVM groups (P<0.001). No significant differences were observed in blastocyst cell numbers among all five groups. CONCLUSIONS: While FSH priming improves the maturation rate of IVM oocytes, FSH or hCG priming does not improve development to the blastocyst stage.

Animals↗

Angiogenesis in embryonic development--a review.

Recent studies, particularly of genetically altered mice, have greatly improved our understanding of the molecular basis underlying the development of the vascular system. Endothelial signalling systems, such as the vascular endothelial growth factor (VEGF)/VEGF receptor system and the angiopoietin/Tie2 system, were identified as central regulators of embryonic angiogenesis. The complex interactions between these and other endothelial signalling systems are beginning to emerge. Placenta (2000), 21, Supplement A, Trophoblast Research, 14, S11-S15.

Angiopoietin-1↗

Commitment of fetal male germ cells to spermatogonial stem cells during mouse embryonic development.

The continuous production of mammalian sperm is maintained by the proliferation and differentiation of spermatogonial stem cells that originate from primordial germ cells (PGCs) in the early embryo. Although spermatogonial stem cells arise from PGCs, it is not clear whether fetal male germ cells function as spermatogonial stem cells able to produce functional sperm. In the present study, we examined the timing and mechanisms of the commitment of fetal germ cells to differentiate into spermatogonial stem cells by transplantation techniques. Transplantation of fetal germ cells into the seminiferous tubules of adult testis showed that donor germ cells, at 14.5 days postcoitum (dpc), were able to initiate spermatogenesis in the adult recipient seminiferous tubules, whereas no germ cell differentiation was observed in the transplantation of 12.5-dpc germ cells. These results indicate that the commitment of fetal germ cells to differentiate into spermatogonial stem cells initiates between embryonic days 12.5 and 14.5. Furthermore, the results suggest the importance of the interaction between germ cells and somatic cells in the determination of fetal germ cell differentiation into spermatogonial stem cells, as normal spermatogenesis was observed when a 12.5-dpc whole gonad was transplanted into adult recipient testis. In addition, sperm obtained from the 12.5- dpc male gonadal explant had the ability to develop normally if injected into the cytoplasm of oocytes, indicating that normal development of fetal germ cells in fetal gonadal explant occurred in the adult testicular environment.

Animals↗

Obstruction and recanalization of the ureter during embryonic development.

Major controversies still exist regarding the terminology, the etiology and the pathogenesis of congenital obstructive diseases of the ureter. To try to provide some additional information to this controversial subject, a comparative study of ureteral development in rat and human embryos, using light and electron microscopy, has been performed. During fetal development we observed and demonstrated the existence of obstructive phenomena, both at the level of the ureterovesical junction (Chwalla's membrane) and along the ureter. At the end of the embryonic period, the ureter undergoes a physiologic recanalization process.

Animals↗

Effect of repeated ACTH-stimulation on early embryonic development and hormonal profiles in sows.

This study was conducted to evaluate the effect of adrenal stimulation by synthetic adrenocorticotrophic hormone (ACTH) on the first 2 days of pregnancy in 22 multiparous sows. The experiment was performed during the second oestrus after weaning and the sows were divided into one control (C-group) and one experiment group (E-group). To determine the time of ovulation, transrectal ultrasonographic examination was performed. E-group sows were treated repeatedly with 0.1 mg/kg bodyweight of synthetic ACTH (tetracosactide) i.v. 4-8h after ovulation and continuing every 6h, until slaughter. Blood samples were collected every second hour from about 12h before expected ovulation until slaughter and were analyzed for cortisol, prostaglandin F(2 alpha) -metabolite, and progesterone (P(4)). All sows were slaughtered approximately 48 h after ovulation and the isthmic part of the oviduct was divided into three equally long segments and flushed separately with phosphate buffered saline (PBS). The uterine horns were also flushed with PBS. The embryos of the E-group sows tended (P=0.056) to have a lower cleavage rate than the embryos of the C-group sows but there was no difference between groups in oviductal transport rate of the embryos. In the E-group, significantly (P<0.05) more sows had only embryos with <20 spermatozoa attached to the ZP compared with the C-group. The plasma concentration of cortisol was significantly higher (P<0.0001) in the E-group sows during the time of treatment while the baseline level of prostaglandin F(2 alpha) -metabolite was significantly lower. The baseline level of progesterone increased in both groups after ovulation but there was no significant difference between the groups. Repeated ACTH-stimulation (1) had no effect on the oviductal transport rate of the embryos, (2) had a negative effect on the embryo development, (3) and caused a changed endocrine profile that might have changed oviductal milieu affecting embryo development.

Adrenocorticotropic Hormone↗

Fluid-dynamical basis of the embryonic development of left-right asymmetry in vertebrates.

Experimental work in developmental biology has recently shown in mice that fluid flow driven by rotating cilia in the node, a structure present in the early stages of growth of vertebrate embryos, is responsible for determining the normal development of the left-right axis, with the heart on the left of the body, the liver on the right, and so on. The role of physics, in particular, of fluid dynamics, in the process is one of the important questions that remain to be answered. We show with an analysis of the fluid dynamics of the nodal flow in the developing embryo that the leftward flow that has been experimentally observed may be produced by the monocilia driving it being tilted toward the posterior. We propose a model for morphogen transport and mixing in the nodal flow and discuss how the development of left-right asymmetry might be initiated.

Animals↗

Effects of the porcine oviduct-specific glycoprotein on fertilization, polyspermy, and embryonic development in vitro.

This study evaluated the effects of porcine oviduct-specific glycoprotein (pOSP) on in vitro fertilization (IVF), polyspermy, and development to blastocyst. Experiment 1 evaluated the effects of various concentrations (0-100 microgram/ml) of purified pOSP on fertilization parameters, including penetration, polyspermy, male pronuclear formation, and mean number of sperm penetrated per oocyte. Experiment 2 examined the ability of an anti-pOSP immunoglobulin G to inhibit the observed effects of pOSP on fertilization parameters. Experiments 3 and 4 examined various concentrations of pOSP (0-100 microgram/ml) on zona pellucida solubility and sperm binding, respectively. Lastly, experiment 5 assessed the effects of various concentrations of pOSP (0-100 microgram/ml) on the in vitro embryo cleavage rate and development to blastocyst. Pig oocytes matured and fertilized in vitro were used for all experiments. An effect of treatment (P < 0.05) was detected for pOSP on penetration, polyspermy, and mean number of sperm per oocyte. Concentrations for pOSP of 0-50 microgram/ml had no effect on sperm penetration rates; however, compared with the control, 100 microgram/ml significantly decreased the penetration rate (74% vs. 41%). Addition of 10-100 microgram/ml significantly reduced the polyspermy rate compared with the control (61% vs. 24-29%). The decrease in polyspermy achieved by addition of pOSP during preincubation and IVF was blocked with a specific antibody to pOSP. No effect of treatment was observed on zona digestion time relative to the control; however, the number of sperm bound to the zona pellucida was significantly decreased by treatment (P < 0.05). Compared with the control, all concentrations of pOSP examined reduced the number of sperm bound per oocyte (45 vs. 19-34). A treatment effect (P < 0.05) was observed for pOSP on embryo development to blastocyst but not on cleavage rates. Addition of pOSP during preincubation and fertilization significantly increased postcleavage development to blastocyst, but a synergistic stimulation on development was not detected when pOSP was included during in vitro culture. These results indicate that exposure to pOSP before and during fertilization reduces the incidence of polyspermy in pig oocytes, reduces the number of bound sperm, and increases postcleavage development to blastocyst.

Animals↗

The embryonic development of the human paraseptal cartilage.

The paraseptal cartilage was first named by Spurgat in 1896. In the late 19th and early 20th century, the paraseptal cartilage of the lower and higher mammals was studied in great detail; however, very few studies were carried out on the human paraseptal cartilage. Over the past 20 years no articles have been published in the English literature on this topic. Despite all the early interest in this topic, several questions still remain unanswered about the origin of paraseptal cartilages, their ultimate fate (i.e. do they disappear or persist?) and their relationship with the vomer, the nasal capsule, and with Jacobson's organ. Eighteen human fetus heads ranging in age from eight to 32 weeks were studied and the literature reviewed. The development of the paraseptal cartilage and its clinical significance are discussed.

Adult↗

Interaction between splotch (Sp) and curly tail (ct) mouse mutants in the embryonic development of neural tube defects.

The mouse mutations splotch (Sp) and curly tail (ct) both produce spinal neural tube defects with closely similar morphology, but achieve this by different embryonic mechanisms. To determine whether the mutants may interact during development, we constructed mice carrying both mutations. Double heterozygotes exhibited tail defects in 10% of cases, although the single heterozygotes do not express this phenotype. Backcrosses of double heterozygotes to ct/ct produced offspring with an elevated incidence of neural tube defects, both spina bifida and tail defects, compared with a control backcross in which Sp was not involved. Use of the deletion allele Sp2H permitted embryos carrying a splotch mutation to be recognised by polymerase chain reaction assay. This experiment showed that only embryos carrying Sp2H develop spina bifida in the backcross with ct/ct, suggesting that the genotype Sp2H/+, ct/ct is usually lethal around the time of birth as a result of severe disturbance of neurulation. The interaction between Sp and ct was investigated further by examining embryos in the backcross for developmental markers of the Sp/Sp and ct/ct genotypes. Sp/Sp embryos characteristically lack neural crest derivatives, such as dorsal root ganglia, and die on day 13 of gestation. Double mutant embryos from the backcross did not exhibit either of these characteristics suggesting that homozygosity for ct does not cause Sp/+ embryos to develop as if they were of genotype Sp/Sp. The angle of ventral curvature of the posterior neuropore region is enhanced in affected ct/ct embryos whereas it was found to be reduced in Sp/Sp embryos compared with their normal littermates.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Fliih, a gelsolin-related cytoskeletal regulator essential for early mammalian embryonic development.

The Drosophila melanogaster flightless I gene is required for normal cellularization of the syncytial blastoderm. Highly conserved homologues of flightless I are present in Caenorhabditis elegans, mouse, and human. We have disrupted the mouse homologue Fliih by homologous recombination in embryonic stem cells. Heterozygous Fliih mutant mice develop normally, although the level of Fliih protein is reduced. Cultured homozygous Fliih mutant blastocysts hatch, attach, and form an outgrowing trophoblast cell layer, but egg cylinder formation fails and the embryos degenerate. Similarly, Fliih mutant embryos initiate implantation in vivo but then rapidly degenerate. We have constructed a transgenic mouse carrying the complete human FLII gene and shown that the FLII transgene is capable of rescuing the embryonic lethality of the homozygous targeted Fliih mutation. These results confirm the specific inactivation of the Fliih gene and establish that the human FLII gene and its gene product are functional in the mouse. The Fliih mouse mutant phenotype is much more severe than in the case of the related gelsolin family members gelsolin, villin, and CapG, where the homozygous mutant mice are viable and fertile but display alterations in cytoskeletal actin regulation.

Actins↗