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PubMed1993

Embryonic and fetal development: fundamental research.

Much progress has been made over the past decades in the development of in vitro techniques for the assessment of chemically induced effects in embryonic and fetal development. In vitro assays have originally been developed to provide information on the mechanism of action of normal development, and have hence more adequately been used in fundamental research. These assays had to undergo extensive modification to be used in developmental toxicity testing. The present paper focuses on the rat whole embryo culture system, but also reviews modifications that were undertaken for the in vitro chick embryo system and the aggregate cultures of fetal rat brain cells. Today these tests cannot replace the existing in vivo developmental toxicity tests. They can, however, be used to screen chemicals for further development or further testing. In addition, these in vitro tests provide valuable information on the mechanisms of developmental toxicity and help to understand the relevancy of findings for humans. In vitro systems, combined with selected in vivo testing and pharmacokinetic investigations in animals and humans, can thus provide essential information for human risk assessment.

Animals↗
PubMed2004

Pre-decondensed sperm head injections into female pronuclei result in chromosomal mingling, zygotic cleavage, and adequate embryonic and fetal development up to delivery of healthy offspring: a novel method of assisted syngamy.

Investigation of the developmental potential post-injection of a pre-decondensed or non-pre-decondensed sperm head into the female pronucleus of a pre-activated oocyte. Rat pre-activated oocytes were treated with intrapronuclear pre-decondensed sperm head injections (IPSHI) (n = 133) or intrapronuclear non-pre-decondensed sperm head injections (INPSHI) (n = 138). All injected oocytes were transferred to pseudopregnant female recipients. Rat IPSHI techniques resulted in the delivery of five healthy offspring. Rat INPSHI techniques did not result in any pregnancies. Rat IPSHI techniques can result in delivery of healthy offspring. Successful performance of human IPSHI techniques might serve as a novel method to manage cases of intracytoplasmic sperm injection failure due to lack of development of male pronucleus or due to failure in pronuclei fusion.

Animals↗
PubMed1995

Differential localisation of UDP-glucuronosyltransferase in kidney during human embryonic and fetal development.

The aim of our study was to localise UDP-glucuronosyltransferase (UDPGT) in the developing mesonephric and metanephric kidneys of the human embryo and fetus, using immunohistochemical methods and an antibody preparation with broad specificity to the human isoforms. In embryonic and early fetal development of the metanephric kidney, UDPGT is located primarily in derivatives of the ureteric bud such as the ureter, pelvis, calyces and collecting ducts. This early predominance of UDPGT to ureteric bud derivatives declines by mid-fetal life: a) as nephrons evolve and develop they become increasingly UDPGT immunoreactive such that in mature metanephric kidney, the proximal tubules are highly UDPGT reactive, with other elements of the nephron also immunopositive (albeit at lower reactivities) and b) with the formation of an immunonegative transitional epithelium in ureter, pelvis and calyces, the reactivity retained in collecting ducts is only a small proportion of the total. The distribution of UDPGT immunoreactivity is relatively uniform in proximal tubular cells throughout development. This is in contrast to collecting ducts where, in fetal life, this reactivity is displaced to apices and bases by intracellular glycogen deposits. Parietal cells of Bowman's capsule are immunoreactive, but glomeruli are negative. In mesonephric kidney, as early as 32 days post-ovulation, tubules and the mesonephric duct are UDPGT immunoreactive and mesonephric immunopositivity overlaps with that in the developing metanephric kidney.

Embryonic and Fetal Development↗
PubMed1994

Phenolsulphotransferase: localization in kidney during human embryonic and fetal development.

The aim of our study was to localize phenolsulphotransferase (PST) in the developing mesonephric and metanephric kidneys of the human embryo and fetus using immunohistochemical methods with an antibody preparation recognizing members of the human phenolsulphotransferase enzyme family. In embryonic and early fetal development of the metanephric kidney, PST is located primarily in derivatives of the ureteric bud such as the ureter, pelvis, calyces and collecting ducts. This predominance declines by mid-fetal life: first, as nephrons evolve and develop they become increasingly PST-immunoreactive such that in mature metanephric kidney, the proximal tubules are highly PST-reactive, with other elements of the nephron also immunopositive (albeit at lower reactivities) and secondly, with the formation of an immunonegative transitional epithelium in ureter, pelvis and calyces, the reactivity retained in collecting ducts is only a small proportion of the total. The distribution of PST immunoreactivity is relatively uniform in proximal tubular cells throughout development, in contrast to collecting ducts, where, in fetal life, this reactivity is displaced to apices and bases by intracellular glycogen deposits. Mesonephric kidney tubules and the mesonephric duct are PST-immunoreactive and although mesonephric immunopositivity overlaps with that in the developing metanephric kidney the renal contribution to sulphation is absent or low at a time when the developing conceptus is most vulnerable to the potential toxic effects of teratogens.

Adult↗
PubMed1984

Expression of cellular oncogenes during embryonic and fetal development of the mouse.

Cellular oncogenes are conserved with great fidelity across a broad span of evolution. This avid conservation suggests possible roles in critical physiologic functions. Little, however, is known about their activity in normal cellular processes. In this study, we examined the expression pattern of eight cellular oncogenes during embryonic and fetal development of the mouse. Five of these genes (c-myc, c-erb, c-Ha-ras, c-src, and c-sis) were expressed at appreciable levels, and four were modulated in a consistent manner during the course of prenatal development.

Age Factors↗
PubMed1996

Localization of large-T oncoprotein during the embryonic and fetal development of transgenic mice.

Oncoproteins are not only involved in the development of tumors but also play a role in physiological regulation in embryonic growth and differentiation. The mechanisms by which regulation is accomplished in embryonic stages differ from postnatal or adult stages. Oncoproteins responsible for tumors in the adult, i.e., products of proto-oncogenes, are prevented from causing tumors in the embryo. If oncogenes are introduced artificially into the embryo. will they be governed by the embryonic regulation described above? To answer this question we used transgenic mice in which the hybrid construct MSV-SV40-Large-T, composed of the Simian-Virus-Oncogene. Large-T with its SV40-promotor and the Moloney Murine Sarcoma Virus (MSV)-enhancer, had been integrated. Under the influence of large-T expression, these animals develop either brain or endocrine pancreas tumors. In the present investigation, we localised large-T expression during development of mouse embryos and fetuses. Interestingly, we saw large-T positive reactions in organ anlagen other than those that later develop tumors. We found large-T antigens in cartilage anlagen, e.g., in ribs and vertebrae, particularly in fetuses of days 14 to 17, and also in a variety of epithelial cells such as in the lung or the choroid plexus. Our results indicate that, as for proto-oncogene products, the effect of an artificially introduced transgenic oncogene product can also be regulated by embryonic cells.

Animals↗
PubMed1995

Inhibin and activin in embryonic and fetal development in ruminants.

Inhibin, activin and follistatin are protein hormones with diverse physiological roles. The involvement of inhibin in the regulation of pituitary FSH production and secretion in adult males and non-pregnant females is well established. However, it is unlikely that inhibin plays a similar role in pregnancy in ruminants. Inhibin and activin molecules show a high degree of structural similarity to potent growth and differentiation factors of the transforming growth factor beta (TGF-beta) superfamily of peptides and their localization in a range of embryonic and fetal tissues indicates that they may thus play a role in development. Furthermore, the demonstration that follistatin is also present in a number of embryonic and fetal tissues and fluids has further implications for the actions of activin to which it binds. The role of inhibin, activin and follistatin in early development has yet to be established since gene knockout experiments have so far proved inconclusive. During mid- and late gestation, high concentrations of inhibin are found in the testes and plasma of male fetuses of sheep and cattle. Inhibin may play a role in regulating pituitary FSH release in late pregnancy, but the very high concentrations of this hormone in ovine fetal testes and in male fetal plasma compared with that observed in the fetal ovary and female fetal plasma has yet to be explained. The recent observation of high concentrations of inhibin, activin and follistatin in amniotic fluid surrounding the fetus is intriguing. Excretion via urine or lung liquid is partly responsible for the presence of these proteins in amniotic fluid. The fetal membranes and the placenta are also possible sources. It remains to be established whether these proteins constitute an inactive pool of secreted hormone or whether they have other actions in this fetal compartment.

Activins↗
PubMed1998

Embryonic and fetal development of structures associated with the cerebro-spinal fluid in man and other species. Part I: The ventricular system, meninges and choroid plexuses.

Little is known about the development of the central nervous system (CNS) in humans. Ethical considerations preclude experimental studies in this field, and as a result most available data on human ontogenesis are descriptive. Comparative anatomic and embryologic studies have demonstrated that the main developmental milestones are conserved across species, and their results can be used to suggest a likely scenario for human development. The development of the ventricles, meninges, and choroid plexuses are discussed in this article. The central cavity of the neural tube is formed during neurulation, which occurs during the fourth gestational week. The first milestone is occlusion of the spinal neurocele (the central canal in the neural tube) shortly after neurulation. This prevents free communication between the ventricular system and the amniotic cavity. The second milestone is development of the meninges, which separate the central nervous system from the rest of the body. The embryonic origin of the meninges varies across species. In birds (and probably in mammals), the spinal meninges are derived from the somitic mesoderm, the brainstem meninges from the cephalic mesoderm, and the telencephalic meninges from the neural crest. Differentiation of the meninges, which involves formation of the subarachnoid space, occurs early, before the cerebrospinal fluid (CSF) begins to flow around the CNS. During ontogenesis, the meninges play a key role in regulating the growth of underlying nervous structures. They induce the formation of the superficial glial limiting layer and stimulate the growth of precursors located in the superficial blastemas of the cerebellum and hippocampus. The choroid plexuses are complex specialized structures that produce most of the CSF. Their epithelium derives from the neural tube epithelium and their mesenchyma from the meninges. Of the many enzymes produced in the choroid plexuses, some reflect the pivotal metabolic role of these structures (alkaline and acid phosphatases, magnesium-dependent ATPase, glucose-6-phosphatase, thiamine pyrophosphatase, adenylate cyclase, oxidoreductase, esterases, hydrolases, cathepsin D, and glutathion S-transferase). The two enzymes that are crucial to the production of CSF are Na+/K+ ATPase and carbonic anhydrase. Inactivation of catecholamines is mediated by catechol-O-methyltransferase and by the monoamine oxidases A and B. The morphology and synthesis profile of the choroid plexuses changes during development, although little is known about these changes in humans.

Animals↗
PubMed1987

A fetuin-related glycoprotein (alpha 2HS) in human embryonic and fetal development.

The human plasma protein, alpha 2HS glycoprotein, has an amino acid composition very similar to that of fetuin, the major protein in fetal calf and lamb serum. Immunohistochemical studies of human fetuses (6-33 weeks gestation) showed that alpha 2HS glycoprotein and fetuin have similar distributions in developing brain and several other tissues, e.g., bone, kidney, gonads, gastrointestinal tract, respiratory and cardiovascular systems. There were notable differences in the liver and thymus in the distribution of the two proteins. Fetuin and alpha 2HS glycoprotein are present in plasma and cerebrospinal fluid of both human and sheep fetuses; their concentrations are reciprocally related: in human plasma and cerebrospinal fluid alpha 2HS glycoprotein concentration is high and fetuin low; the reverse is the case in sheep fetuses. Estimates of the concentration of alpha 2HS glycoprotein in human fetal cerebrospinal fluid and plasma were obtained. It is suggested that alpha 2HS glycoprotein may play a role in developing tissues, especially in the human fetus, similar to that of fetuin in other species.

Blood Proteins↗
PubMed2003

The role of essential trace elements in embryonic and fetal development in livestock.

This review addresses the concept that essential trace minerals play a vital role in many enzymatic and metabolic pathways that are critical for conceptus development during pregnancy in livestock species. The conceptus relies entirely on the maternal system for a sufficient supply of trace minerals and other nutrients needed for normal development. If this supply is inadequate, growth and/or health of the conceptus can be affected adversely, and many of these effects carry over into the neonatal period. Information, accumulated in our laboratory and presented herein, indicates that zinc, copper and manganese are among the trace minerals that have the greatest impact on reproduction. For example, levels of zinc, copper and manganese were several fold greater in the conceptus than in other reproductive tissues, indicating that the conceptus preferentially accumulates these minerals, an action that may be important for conceptus development, growth and survival. Moreover, some recent results indicate that increasing the biological availability of zinc, copper and manganese, by attachment to short peptide chains (i.e., proteinated trace minerals) can enhance reproductive performance of swine. Mineral concentrations in conceptuses from female pigs consuming proteinated trace minerals were greater than those from females that consumed only inorganic mineral salts. Elucidating the mechanisms whereby conceptus development and survival are enhanced by essential trace minerals may lead to development of specific feeding programs to increase the number and health of offspring at parturition, thereby allowing for further improvements in production efficiency in animal agriculture.

Animal Husbandry↗
PubMed1992

Toxicology of maternally ingested trichloroethylene (TCE) on embryonal and fetal development in mice and of TCE metabolites on in vitro fertilization.

Trichloroethylene (TCE), an industrial solvent, is a soil and ground water contaminant found across the United States. The metabolism and carcinogenic potential of TCE have been studied extensively in the past 15 years yet there is little information on the chemical's possible effects on reproduction. No reference to the reproductive effects in mice of TCE by oral administration exists in the literature. In this study, female B6D2F1 mice were gavaged from Days 1 to 5, 6 to 10, or 11 to 15 (Day 1 = vaginal plug) with TCE in corn oil at 0, 1/10, and 1/100 of the oral LD50. Weights of mice were recorded and the livers and kidneys were weighed and preserved in 10% buffered formalin. Litters were counted, sexed, weighed, and measured for crown-rump length until weaning on Day 21 and some animals were allowed to develop to 6 weeks of age. At this time, a minimum of two litters from each dose were killed and gonads removed, weighed, and preserved in Bouin's fixative. Litters were also assessed for developmental abnormalities. No maternal or reproductive effect of TCE was seen at either dose level. TCE, administered consecutively on Days 1 to 5, 6 to 10, and 11 to 15 of pregnancy, does not appear to be a potential reproductive toxicant up to 1/10 the oral LD50. In a second series of studies, TCE and its metabolites dichloroacetic acid (DCA), trichloroacetic acid (TCAA), and trichloroethanol (TCOH) were added to culture media to assess the toxic effects on in vitro fertilization (IVF) in mice.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1992

Prenatal expression of the growth hormone (GH) receptor/binding protein in the rat: a role for GH in embryonic and fetal development?

Although fetal growth is generally considered to be independent of pituitary growth hormone (GH), it is possible that pituitary GH plays a modulatory role in organ development or that a GH-like substance of non pituitary origin may influence fetal growth through the GH receptor. Accordingly, we have used immunohistochemistry, northern blot analysis, the reverse transcriptase-polymerase chain reaction and solution hybridization to study the ontogeny of the GH receptor/binding protein (BP) from the 12-day-old embryo (E12) to the E18 rat fetus. GH receptor/BP immunoreactivity was observed in all major organ systems of the E18 rat fetus and was not preferentially associated with any germ layer derivative. A general increase in GH receptor/BP immunoreactivity was evident from E12 to E18, with a marked increase occurring between E16 and E18. Hemangioblastic tissue was, however, strongly or intensely immunoreactive at all stages of development, as was the placenta. Most noteworthy of the other tissues expressing GH receptor/BP immunoreactivity by day 18 were skeletal and smooth muscle, chondroprogenitor cells, epithelial lining cells, neuronal ganglia, ependymal cells and the adrenal cortex. In the placenta, the most prominent immunoreactivity was associated with decidual cells. Total RNA was isolated from E12 to E18 rat fetuses and adult rat liver. Northern hybridization with a 35S-labelled rat GH receptor cRNA probe revealed that 3.9 kb and 1.2 kb transcripts complementary to the rat GH receptor riboprobe are present from at least E16. The existence of GH receptor mRNA at E12 and E14 was demonstrated by the polymerase chain reaction.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1989

Effects of inhalation exposure to carbon disulfide and its combination with hydrogen sulfide on embryonal and fetal development in rats.

Pregnant rats were exposed to 0, 100, 200, 400 or 800 ppm of carbon disulfide (CS2), 100 ppm of hydrogen sulfide (H2S) alone or in combination with 400 and 800 ppm CS2, 6 h/d during days 6-20 of gestation. Maternal reproduction and fetal parameters were evaluated on gestational day 21. Treatment with 100 or 200 ppm CS2 or with 100 ppm H2S caused no maternal toxicity or adverse effects on the developing embryo or fetus. Exposure to 400 or 800 ppm CS2 resulted in a low incidence of club foot and in a significant reduction of maternal weight gain. Significant increases in unossified sternebrae occurred at 800 ppm CS2 and reduction of fetal body weight at 400 and 800 ppm CS2. The latter effect was enhanced by combination with 100 ppm H2S. These results support the conclusion that, at levels of exposure associated with maternal toxicity, CS2 leads to an increase in incidence of club foot and to fetal toxicity which is enhanced by simultaneous exposure to H2S.

Administration, Inhalation↗