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PubMed1991

Growth and motor development in fetuses of women with type-1 diabetes. II. Emergence of specific movement patterns.

In 20 women with type-1 diabetes, the emergence of fetal movement patterns was studied using real-time ultrasound. One-hour recordings were made once a week between the 7th and 17th week of gestation. Data were compared to those obtained in uncomplicated pregnancy. The diabetic women were being treated with continuous subcutaneous insulin infusion (CSII) therapy. Except for breathing movements, there was a 1-2 week delay in the first appearance of all movement patterns which normally emerge during the first 12 weeks of pregnancy. Breathing movements were observed for the first time at an earlier age than in the control fetuses (P less than 0.02). When the emergence of frequently occurring movement patterns was plotted against fetal crown-rump length, which is usually smaller in diabetic pregnancy, there was still a general delay in comparison with the control group. The delay in motor development therefore does not run completely parallel with the delay in growth. This indicates the possible existence of a specific diabetes-related influence on the functional development of the embryonic and fetal nervous system. Hyperglycaemia, for example, may be responsible, as the delay in the emergence of fetal general movements was most profound in the women whose periconceptional quality of glucose control was poor.

Anthropometry↗
PubMed2002

The role of growth hormone in fetal development.

Studies across several species, particularly the mouse, show that growth hormone (GH, somatotrophin) is an important determinant of litter size, and to a lesser extent, of birth length. GH acts at all stages of development, from ovulation through preimplantation development to the late fetus, with actions on both embryo/fetus and mother contributing to successful fetal development. The fact that these are not more obvious in vivo is likely a result of redundancy of cytokine hormone action, particularly in relation to prolactin, which shares common actions and receptor locations with GH.

Animals↗
PubMed1996

Insulin-like growth factor I promotes growth selectively in fetal sheep in late gestation.

Insulin-like growth factor I (IGF-I) is required for normal fetal growth and skeletal maturation in late gestation, because null mutations of the IGF-I gene in mice reduce fetal weight and retard ossification of bones. To determine if, conversely, increased abundance of IGF-I promotes fetal growth and skeletal maturation, fetal sheep were infused intravascularly with recombinant human IGF-I (n = 7) (26 +/- 3 micrograms. h-1.kg-1) from 120 to 130 days gestation and compared with controls (n = 15). IGF-I infusion increased plasma IGF-I concentrations by 140% (P = 0.002) and weights of fetal liver, lungs, heart, kidneys, spleen, pituitary, and adrenal glands by 16-50% (P < 0.05). Weights and/or lengths of the fetus, placenta, gastrointestinal tract, individual skeletal muscles, and long bones were unchanged by IGF-I. However, IGF-I increased the percentage of proximal epiphyses of long bones present (P < 0.05) and their cross-sectional areas by 15 to 38% (P < 0.05). These results show that IGF-I promotes growth of major fetal organs, endocrine glands, and skeletal maturation in vivo, consistent with IGF-I actively controlling and not merely facilitating fetal growth. The variable response of different tissues may partly reflect tissue specificity in growth requirements for additional factors.

Animals↗
PubMed1994

Late appearance of a type I alveolar epithelial cell marker during fetal rat lung development.

Recent studies in fetal lung using immunological and molecular probes have revealed type I and type II cell phenotypic markers in primordial lung epithelial cells prior to the morphogenesis of these cell types. We have recently developed monoclonal antibodies specific for adult type I cells. To evaluate further the temporal appearance of the type I cell phenotype during alveolar epithelial cell ontogeny, we analyzed fetal lung development using one of our monoclonal antibodies (mAb VIII B2). The epitope recognized by mAb VIII B2 first appears in the canalicular stage of fetal lung development, at approx. embryonic day 19 (E19), in occasional, faintly stained tubules. Staining with this type I cell probe becomes more intense and more widespread with increasing gestational age, during which time the pattern of staining changes. Initially, all cells of the distal epithelial tubules are uniformly labelled along their apical and basolateral surfaces. As morphological differentiation of the alveolar epithelium proceeds, type I cell immunoreactivity appears to become restricted to the apical surface of the primitive type I cells in a pattern approaching that seen in the mature lung. We concurrently analyzed developing fetal lung with an antiserum to surfactant apoprotein-A (alpha-SP-A). Consistent with the findings of others, labeling of SP-A was first detectable in scattered cuboidal cells at E18. Careful examination of the double-labeled specimens suggested that some cells were reactive with both the VIII B2 and SP-A antibodies, particularly at E20. Confocal microscopic analysis of such sections from E20 lung confirmed this impression. Three populations of cells were detected: cells labeled only with alpha-SP-A, cells labeled only with mAb VIII B2, and a smaller subset of cells labeled by both.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed2001

Cryopreservation of ICR mouse oocytes: improved post-thawed preimplantation development after vitrification using Taxol, a cytoskeleton stabilizer.

OBJECTIVE: To establish an effective cryopreservation method. DESIGN: In vitro model study. SETTING: Infertility Medical Center, Pochon CHA University. ANIMAL(S): Four-week-old ICR mice superovulated with pregnant mare serum gonadotropin (PMSG) and human chorionic gonadotropin. INTERVENTION(S): Vitrified-thawed oocytes were fertilized and subsequently cultured in vitro. MAIN OUTCOME MEASURE(S): Post-thawed development, chromosome/spindle normalities, and blastocyst quality. RESULT(S): More cumulus-enclosed oocytes were fertilized and developed to the 8-cell stage after vitrification and thawing than denuded oocytes. However, cryopreserved oocytes of both types had lower spindle and chromosome normalities than fresh oocytes, which resulted in reduced developmental competence after thawing. The addition of 1 microM of Taxol, a cytoskeleton stabilizer, to vitrification solution greatly promoted the blastocyst formation of vitrified-thawed oocytes, compared with no addition (24.0% vs. 58.6%). No difference in blastocyst quality, which was evaluated by blastomere and inner cell mass cell numbers and inner cell mass cell per trophoblast ratio, was found between fresh oocytes and oocytes vitrified with Taxol. CONCLUSION(S): A vitrification solution consisting of 5.5 M ethylene glycol, 1.0 M sucrose, 10% fetal bovine serum, and 1 microM Taxol greatly improved post-thawed development of vitrified oocytes.

Animals↗
PubMed1986

New perspectives in early human development.

A new hypothesis is presented concerning the maturation of the human nervous system and the duration of pregnancy. It has been found that many neural functions characteristic of the fetus continue after birth until a major transformation takes place at the end of the second postnatal month. From a comparative perspective the duration of pregnancy in man is relatively short among primates. An explanation may be found in metabolic constraint. The special energy demands of a large fetal nervous system and the build-up of a subcutaneous fat deposit may have prevented a proportional prolongation of pregnancy during evolution of hominids.

Animals↗
PubMed1998

Smad2 signaling in extraembryonic tissues determines anterior-posterior polarity of the early mouse embryo.

Smad proteins transmit TGFbeta signals from the cell surface to the nucleus. Here we analyze Smad2 mutant embryos created using ES cell technology. Smad2 function is not required for mesoderm production per se, but, rather unexpectedly, in the absence of Smad2 the entire epiblast adopts a mesodermal fate giving rise to a normal yolk sac and fetal blood cells. In contrast, Smad2 mutants entirely lack tissues of the embryonic germ layers. Smad2 signals serve to restrict the site of primitive streak formation and establish anterior-posterior identity within the epiblast. Chimera experiments demonstrate these essential activities are contributed by the extraembryonic tissues. Thus, the extraembryonic tissues play critical roles in establishing the body plan during early mouse development.

Animals↗
PubMed2006

De novo methylation, long-term promoter silencing, methylation patterns in the human genome, and consequences of foreign DNA insertion.

This chapter presents a personal account of the work on DNA methylation in viral and mammalian systems performed in the author's laboratory in the course of the past 30 years. The text does not attempt to give a complete and meticulous account of the work accomplished in many other laboratories; in that sense it is not a review of the field in a conventional sense. Since the author is also one of the editors of this series of Current Topics in Immunology and Microbiology on DNA methylation, to which contributions by many of our colleagues in this field have been invited, the author's conscience is alleviated that he has not cited many of the relevant and excellent reports by others. The choice of viral model systems in molecular biology is well founded. Over many decades, viruses have proved their invaluable and pioneering role as tools in molecular genetics. When our interest turned to the demonstration of genome-wide patterns of DNA methylation, we focused mainly on the human genome. The following topics in DNA methylation will be treated in detail: (1) The de novo methylation of integrated foreign genomes; (2) the long-term gene silencing effect of sequence-specific promoter methylation and its reversal; (3) the properties and specificity of patterns of DNA methylation in the human genome and their possible relations to pathogenesis; (4) the long-range global effects on cellular DNA methylation and transcriptional profiles as a consequence of foreign DNA insertion into an established genome; (5) the patterns of DNA methylation can be considered part of a cellular defense mechanism against foreign or repetitive DNA; which role has food-ingested DNA played in the elaboration of this mechanism? The interest in problems related to DNA methylation has spread-like the mechanism itself-into many neighboring fields. The nature of the transcriptional programs orchestrating embryonal and fetal development, chromatin structure, genetic imprinting, genetic disease, X chromosome inactivation, and tumor biology are but a few of the areas of research that have incorporated studies on the importance of the hitherto somewhat neglected fifth nucleotide in many genomes. Even the fly researchers now have to cope with the presence of this nucleotide, in however small quantities it exists in the genome of their model organism, at least during embryonal development. The bulk of the experimental work accomplished in the author's laboratory has been shouldered by many very motivated undergraduate and graduate students and by a number of talented postdoctoral researchers. Their contributions are reflected in the list of references in this chapter. We have also had the good luck to receive funding through a number or organizations as acknowledged.

Adenoviridae↗
PubMed2001

Perinatal development of endothelial nitric oxide synthase-deficient mice.

The purpose of this study was to evaluate the influence of endothelial nitric oxide synthase (eNOS) deficiency on fetal growth, perinatal survival, and limb development in a mouse model with a targeted mutagenesis of the Nos3 gene. Wild-type (Nos3+/+) and eNOS-deficient fetuses (Nos3-/-) were evaluated on Gestational Day (E)15 and E17, and newborn pups were observed on Day 1 of life (D1). The average term duration of pregnancy was 19 days. For the evaluation of postnatal development, a breeding scheme consisting of Nos3+/- x Nos3+/- and Nos3-/- x Nos3-/- mice was established, and offspring were observed for 3 wk. Southern blotting was used for genotyping. No significant differences in fetal weight, crown-rump lengths (CRL), and placental weight were seen between Nos3+/+ and Nos3-/- fetuses on E15. By E17, Nos3-/- fetuses showed significantly reduced fetal weights, CRL, and placental weights. This difference in body weight was also seen throughout the whole postnatal period. In pregnancies of Nos3-/- females, the average number of pups alive on D1 was significantly decreased compared to either E15 or E17. Placental histology revealed no abnormalities. On E15, E17, and D1, Nos3(-/-) fetuses demonstrated focal acute hemorrhages in the distal limbs in 0%, 2.6%, and 5.7%, respectively, of all mutant mice studied on the respective days. Bone measurements showed significantly shorter bones in the peripheral digits of hindpaws of Nos3-/- newborns. We conclude mice deficient for eNOS show characteristically abnormal prenatal and postnatal development including fetal growth restriction, reduced survival, and an increased rate of limb abnormalities. The development of this characteristic phenotype of eNOS-deficient mice dates back to the prenatal development during the late third trimester of pregnancy.

Animals↗
PubMed2000

Post-implantation development of mouse androgenetic embryos produced by in-vitro fertilization of enucleated oocytes.

We report here on the precise ability of mouse androgenetic embryos produced by in-vitro fertilization of enucleated oocytes to develop to day 9.5 of gestation when cultured with M16 and CZB media. Androgenetic embryos cultured with CZB rather than M16 medium developed to the blastocyst stage in a more significant proportion (56.6% versus 45.0%, P < 0.001). However, after cavitation, the rate of cell proliferation of androgenetic embryos cultured with CZB medium was significantly decreased (P < 0.05). Embryo transfer experiments showed that blastocysts cultured with M16 medium were superior to those cultured with CZB medium in their ability to develop to 9.5-day-old fetuses (28.1% versus 11.1%, P < 0.001). These results showed that the present procedure for producing androgenetic mouse embryos is reliable and that M16 medium is superior for culturing the embryos. Fetal sexing by polymerase chain reaction (PCR) also demonstrated that both XX and XY embryos develop to 9.5-day fetuses at theoretical rates (1:2). This is the first finding that mouse XX androgenones survive after implantation.

Animals↗
PubMed2001

Fetal growth, early postnatal growth and motor development in Pakistani infants.

BACKGROUND: Studies have often compared the postnatal motor development of 'small' versus 'normal' newborns. Not much is known about the associations between a broad spectrum of size at birth and motor development. The effect of early postnatal growth on motor development is little researched. Growth failure in terms of shortness and thinness should be differentiated, but not many studies have the data for this analysis. METHODS: This is a longitudinal study of infants born in Lahore, Pakistan, between 1984 and 1987. Age at commencement of independent walking and age at 'building a 3-cube tower' were taken as indicators of gross and fine motor development, respectively. Size at birth was captured by length and thinness as continuous variables; postnatal growth from birth to 6 months of age was measured by changes in length and thinness. Adjustment for covariates and handling of censored cases were performed by generalized log gamma regression. RESULTS: Thinness at birth and postnatal stunting and wasting had a linear, inverse association with gross motor development (each P < 0.05). Birth length had a non-linear, inverse association with this outcome (P < 0.05). Birth length, thinness at birth and postnatal wasting had a linear, inverse association with fine motor development (each P < 0.05). CONCLUSION: Both fetal and early postnatal growth over a broad spectrum may affect infants' motor development. It is not just the babies who were very small at birth that suffered. Birth length appeared to be more influential than other anthropometric indicators.

Child Development↗
PubMed1997

Postnatal development and behaviour of Wistar rats after prenatal toluene exposure.

Pregnant Wistar rats were treated with different concentrations of toluene by inhalation (300, 600, 1000 and 1200 ppm) from day 9 to day 21 of pregnancy for 6 h a day in a whole-body inhalation chamber (controls inhaled fresh air only). From day 22, rats were kept single-caged and were allowed to deliver. Besides a detailed evaluation of the physical development of the offspring we performed the following tests: forelimb-grasp reflex, righting reflex, cliff-drop aversion reflex, maintainance of balance on a rotating rod, measurement of locomotor activity and learning ability in a discrimination learning test. A toluene exposure of 1200 ppm resulted in a reduced body weight of rat dams and offspring and a higher mortality until weaning. The physical development (incisor eruption, eye opening and vaginal opening) was retarded in this group. There were no clear-cut and concentration-dependent differences in the development of reflexes, rota rod performance and locomotor activity between the offspring of animals exposed to toluene and the controls. Likewise, no effects were found on learning ability in the operant conditioning task. Compared to the controls there were no differences in mating, fertility and pregnancy indexes in the F1-generation. The tests performed have provided no evidence that toluene exposures < or = 1200 ppm induce adverse effects on the behaviour of rat offspring exposed during late embryonic and fetal development.

Animals↗
PubMed2004

Preimplantational embryo development and incidence of blastomere apoptosis in bovine somatic cell nuclear transfer embryos reconstructed with long-term cultured donor cells.

This study was performed to investigate whether types and/or age of donor cells affect preimplantational embryo development and the incidence of apoptosis in bovine somatic cell nuclear transfer (SCNT) embryos. Bovine fetal or adult ear fibroblasts were isolated, cultured in vitro and categorized into fresh or long-term cultured cells in terms of population doublings (PD): in fetal fibroblasts, <16 being considered fresh and >50 being long-term cultured; in adult ear fibroblasts, <16 being considered fresh and >30 being long-term cultured. Bovine oocytes from slaughterhouse ovaries were matured in TCM-199, enucleated and reconstructed by SCNT. The reconstructed oocytes were fused, chemically activated, and cultured in modified synthetic oviduct fluid (mSOF) at 39 degrees C in a humidified atmosphere of 5% CO(2) air for 7 days. The early development of SCNT embryos was monitored under a microscope and the quality of blastocysts was assessed by differential counting of inner cell mass (ICM) and trophectoderm (TE) cells and by apoptosis detection in blastomeres using a terminal deoxynucleotidyl transferase-mediated d-UTP nick end-labeling (TUNEL) assay. As results, types and/or age of donor cells did not affect the rate of blastocyst formation and the number of ICM and TE cells. However, a significant increase in apoptotic blastomeres was observed in SCNT embryos reconstructed with long-term cultured fetal or adult ear fibroblasts compared to those in SCNT embryos derived from fresh fetal or adult ear fibroblasts. In conclusion, these results indicated that the long-term culture of donor cells caused increased the incidence of apoptosis in bovine SCNT embryos but did not affect the developmental competence and the cell number of blastocysts.

Animals↗
PubMed1998

Intracellular divalent cation homeostasis and developmental competence in the hamster preimplantation embryo.

The intracellular magnesium and calcium ion concentrations of in vivo-developed 2-cell hamster embryos were measured using ratiometric fluorometry. Intracellular magnesium and calcium ion concentrations were found to be 0.369 +/- 0.011 mM and 129.3 +/- 7.5 nM respectively. Culture of 1-cell hamster embryos for 24 hr to the 2-cell stage in control medium containing 0.5 mM magnesium and 2.0 mM calcium resulted in approximately a threefold increase to 343.5 +/- 8.0 nM in intracellular calcium ion concentration, while magnesium ion levels were not altered (0.355 +/- 0.007 mM). Increasing medium magnesium concentrations to 2.0 mM significantly increased intracellular magnesium ion concentrations of cultured 2-cell embryos with a concomitant reduction in intracellular calcium ion concentrations. Furthermore, increasing the medium magnesium concentration to 2.0 mM significantly increased development of 1-cell embryos collected at either 3 or 9 hr post-egg activation to the morula/blastocyst and blastocyst stages. Resultant blastocysts had an increased total cell number and increased development of the inner cell mass. Most important, however, culture with 2.0 mM magnesium increased the fetal potential of cultured 1-cells twofold. Therefore, because highest rates of development were observed in a medium that resulted in reduced intracellular calcium ion concentrations, it appears that altered calcium homeostasis is associated with impaired developmental competence of 1-cell embryos in culture.

Animals↗
PubMed1998

Evaluation of early embryonic loss induced by tributyltin chloride in rats: phase- and dose-dependent antifertility effects.

In our previous study, tributyltin chloride (TBTCl) on days 0-7 of pregnancy was found to produce implantation failure in rats. The objective of the present study was to determine the susceptible period for the antifertility effect of TBTCl in rats. Inseminated females were orally administered TBTCl at 8.1, 16.3, or 32.5 mg/kg on days 0-3 of pregnancy, or at 8.1, 16.3, 32.5, or 65.1 mg/kg on days 4-7 of pregnancy. Pregnancy outcome was determined on day 20 of pregnancy. Dosing with TBTCl on days 0-3 of pregnancy at 16.3 mg/kg and higher produced a significant increase in the rate of implantation failure. Dosing with TBTCl on days 4-7 of pregnancy caused a significant increase in the incidence of postimplantation loss at 16.3 mg/kg and higher in females with implantations. No increase in the incidence of fetal malformations was found in any TBTCl-treated groups. It could be concluded that the susceptibility to and manifestation of the antifertility effects of TBTCl vary with the gestational stage at the time of administration.

Administration, Oral↗
PubMed1992

Primitive neuroectodermal tumors after prophylactic central nervous system irradiation in children. Association with an activated K-ras gene.

Three patients had supratentorial malignant brain tumors 7 to 9 years after prophylactic central nervous system (CNS) treatment for acute lymphocytic leukemia or malignant T-cell lymphoma. Therapy was administered at the age of 3 to 8 years and included cranial irradiation (total dose, 1800 to 2400 cGy) and intrathecal methotrexate. The brain tumors had histologic and immunohistochemical features of primitive neuroectodermal tumors (PNET), including neuroblastic rosettes, rhythmic arrangement of tumor cells, and immunohistochemical expression of glial, and in one patient neuronal, marker proteins. Using polymerase chain reaction-mediated DNA amplification from paraffin-embedded tissues and subsequent DNA sequence analysis, an activating point mutation was detected in the K-ras protooncogene in one tumor. This mutation was a G to A transition in position 2 of codon 12, substituting aspartate (GAT) for glycine (GGT). This type of mutation has not been observed before in human brain tumors, but it is frequent in radiation-induced murine lymphomas. These observations suggest that PNET can be induced after completion of the embryonal and fetal development of the human CNS. Oncogene-activating point mutations may represent a pathogenetic mechanism involved in the genesis of radiation-induced brain tumors.

Aspartic Acid↗
PubMed2001

Cardiovascular physiology during fetal development and implications for tissue engineering.

Shear stress in fluid dynamics has a well-known impact on vascular cell morphology, proliferation, orientation, and the organization and composition of extracellular matrix. There is an increasing interest in the field of tissue engineering to use defined shear stress in bioreactors for tissue conditioning and guided tissue formation. Especially for cardiovascular structures like heart valves or blood vessels, the type and appropriate amount of shear stress needed to improve tissue formation remains speculative. The authors believe that fetal-like conditions may be more optimal for new tissue formation in a bioreactor. The purpose of this review is to delineate parameters of cardiovascular physiology during embryonic and fetal development that may have a potential impact on the design and setting of bioreactors for cardiovascular tissue engineering.

Biomedical Engineering↗