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Fetal hemoglobin (HbF) synthesis in baboons, Papio cynocephalus. Analysis of fetal and adult hemoglobin synthesis during fetal development.

Fetal hemoglobin (HbF) and adult hemoglobin (HbA) synthesis was studied in fetal baboons, Papio cynocephalus, to determine the normal pattern of hemoglobin production during fetal development. Fetuses ranging from 53 to 180 days gestation (term gestation 184 days) were used. Erythroid cells were incubated with 3H-L-leucine, and the rates of globin chain synthesis and the distribution of radioactivity into hemoglobin intermediates and completed hemoglobin molecules were determined. Gamma chain synthesis accounted for approximately 97% of the total nonalpha chain synthesis up to 140 days gestation; beta chain synthesis accounted for the remainder. After 140 days gestation, approximately equal quantities of gamma and beta chain were synthesized in the bone marrow. Prior to 140 days gestation, total alpha chain synthesis was 30% greater than total non-alpha chain synthesis, while there was balanced chain synthesis after 140 days gestation. During the period of excess alpha chain synthesis, fetal erythrocytes contained a large pool of alpha-hemoglobin (alpha chain with heme attached) molecules uncombined with beta or gamma chains. In view of the possibility that alpha chains may have a lower affinity for gamma chains than beta chains, excess alpha chain synthesis may be required to maintain low levels of free gamma chains.

Animals↗

Placental hormones, nutrition, and fetal development.

Fetal growth retardation due to maternal malnutrition is widespread especially in the Third World. Little is known about the mechanisms that regulate the growth of the fetus and placenta during protein malnutrition. It is known that the placental size and levels of circulating placental hormones such as human chorionic gonadotrophins (hCG), human placental lactogen (hPL), and estrogens are affected by the nutritional status of the mother. There is suggestive evidence that during malnutrition, hPL may increase lipolysis and exert a glucose sparing effect in the mother, thereby promoting glucose availability to the fetus. We have studied the influence of dietary protein deficiency on the binding of dexamethasone to the specific cytosol receptors in adult and fetal tissues. A low protein diet in adult male rats is associated with a decrease in dexamethasone binding to liver cytosol receptors. On the other hand, protein deprivation in pregnant female rats leads to an increase in dexamethasone binding to liver cytosol receptors of both the mother and fetus. However, the influences of maternal protein deprivation on dexamethasone receptors in the fetal liver and lungs are not similar. At 21 days gestation the binding of dexamethasone to fetal lung receptors of protein-deficient mothers is lower than that in the controls. These differences at a critical time in the fetal lung development indicate that a fall in receptors for dexamethasone may lead to impaired phospholipid synthesis in fetuses of protein-deficient mothers and point to the importance of nutritional factors in the biochemistry of fetal development.

Animals↗

Physiological importance of system A-mediated amino acid transport to rat fetal development.

Fetal growth and development are dependent on the delivery of amino acids from maternal amino acid pools to the fetal blood. This is accomplished via transfer across the apical and basal plasma membrane of the placental syncytiotrophoblast. The aim of this study was to determine whether inhibition of system A (amino acid transporter) was associated with a decrease in fetal weight in the rat. System A is a ubiquitous Na(+)-dependent amino acid transporter that actively transports small zwitterionic amino acids. In brief, system A was inhibited by infusing a nonmetabolizable synthetic amino acid analog, 2-(methylamino)isobutyric acid from days 7-20 of gestation. On day 20, the rats were killed and tissues (maternal liver, fetuses, and placentas) were collected for analysis. The degree of system A inhibition was determined, as was the impact of said inhibition on fetal and maternal weights, system A-mediated placental transport, and placental system A-mediated transporter expression. Our results suggest that when system A is inhibited, fetal weight is diminished [control group: -3.55 +/- 0.04 g (n = 113), experimental group: -3.29 +/- 0.04 g (n = 128)], implying an integral role for system A transport in fetal growth and development in the rat.

Amino Acid Transport System A↗

[Fetal development and fetal heart function analyzed by M-mode echocardiography].

Real time-directed M-mode echocardiograms were obtained for 215 echographically evaluated AFD fetuses between 24 and 41 weeks of gestation to assess the cardiac function of normally growing human fetuses. The results are as follows. 1. A significant linear correlation exists between gestational weeks and end-diastolic wall thickening of RV, IVS and LV, respectively (RVAWTd, IVSTd and LVPWTd). 2. There is no significant difference among three (RVAWTd, IVSTd and LVPWTd) at any time from 24 to 41 weeks. 3. Percent systolic wall thickening for RV, IVS and LV are 39 +/- 24, 68 +/- 37 and 41 +/- 16(%), respectively. 4. Motion of IVS towards left ventricular free wall during systole, no septal motion or paradoxical movement (systolic septal anterior motion) is seen in 62, 35 and 3% of cases, respectively. 5. Paradoxical movement of IVS is seen in the case in which the RV/LV wall exclusion ratio is extremely low.

Echocardiography↗

Hypertrophy and hyperplasia of bovine fetal tissues during development: fetal liver insulin-like growth factor I mRNA expression.

Tissue growth of crossbred fetal beef calves was examined by measuring RNA, DNA, and protein concentrations in liver, heart, and biceps femoris. Furthermore, liver insulin-like growth factor I (IGF-I) mRNA expression and mRNA species size during fetal development was observed. Tissue samples were collected from six fetuses every 42 d of gestation, from d 106 to d 274. In the liver, protein and DNA concentrations decreased, whereas RNA levels remained constant throughout fetal growth. The RNA/DNA and protein/DNA ratios in liver increased with fetal age. Heart DNA and RNA levels decreased, whereas protein concentration and protein/DNA ratios increased with fetal age. Protein and protein/DNA ratios decreased in biceps tissue, whereas DNA and RNA concentrations were constant. IGF-I mRNA was seen at 4.4, 2.5, and 1.2 kb in adult and 4.4, 2.5, and 1.7 kb in fetal bovine liver. Relative expression of liver IGF-I mRNA did not vary during fetal development. The current study shows that during the last 2 and 3 mo of gestation, heart and liver were undergoing hypertrophic growth, whereas biceps tissue did not exhibit the same trend. Elevated ratios of RNA and protein to DNA in liver above that of the heart and biceps suggest extensive hepatic cellular hypertrophy as well as increased transcriptional and translational activity. Insulin-like growth factor I mRNA levels were not related to the changes in RNA, DNA, and protein seen in hepatic tissue.

Animals↗

Corticotrophin-releasing factors in the hypothalamus of the developing fetal sheep.

Corticotrophin-releasing hormone (CRH) and arginine vasopressin (AVP) are secreted from the hypothalamic median eminence to elicit the secretion of ACTH from the pituitary corticotrophs. During fetal development there is progressive maturation of the hypothalamic-pituitary-adrenal axis, manifest as increasing plasma ACTH and cortisol concentrations, which in species such as sheep culminates in the onset of birth. However, the precise nature of the hypothalamic signal controlling fetal pituitary ACTH secretion remains poorly understood. To investigate the ontogeny of this hypothalamic signal, the present study examined immunoreactive and bioactive ACTH-releasing factors in the developing fetal sheep hypothalamus. Immunoreactive CRH and AVP were measured by radioimmunoassay in extracts of hypothalami taken at day 70, day 100, and day 130 gestation (term = 145 days). There was a progressive and significant (P < 0.01) increase in hypothalamic CRH and AVP concentrations which was particularly marked between d100 and d130 gestation. AVP was always present in higher concentrations that CRH, although this difference was significantly reduced by day 130 gestation as the result of a large increase in the content of CRH relative to AVP. Sephadex G50 chromatography revealed that immunoreactive CRH and AVP in hypothalamic extracts existed as single molecular forms corresponding to synthetic peptides at each gestational age. In addition, these immunoreactive forms of CRH and AVP possessed significant ACTH-releasing bioactivity as measured in primary cultures of adult sheep anterior pituitary cells. Furthermore, significant bioactivity was present in high and low molecular weight fractions eluted after chromatography which did not contain any CRH or AVP immunoreactivity.(ABSTRACT TRUNCATED AT 250 WORDS)

Adrenocorticotropic Hormone↗

The physiological role of estriol during human fetal development is to act as antioxidant at lipophilic milieus of the central nervous system.

Estriol is the most abundant estrogen during human pregnancy, it has a low estrogenic effect and its physiological role is not known. Recently it has been shown that estrogens have antioxidant and free-radical scavenging activity; in free form estrogens are lipophilic, here it is proposed that the role of estriol is to act as antioxidant at level of the developing fetal central nervous system, whose tissues are substantially lipid in nature.

Antioxidants↗

An in situ-hybridization study of the localization of retinol-binding protein and transthyretin messenger RNAs during fetal development in the rat.

The patterns, and the cellular sites, of expression of the genes for retinol-binding protein (RBP) and transthyretin (TTR) were studied during early- to mid-stages of rat embryogenesis. In situ hybridization of single-stranded RNA probes to rat embryo sections revealed specific sites at which RBP messenger RNA (mRNA) and TTR mRNA were localized in rat conceptuses from 7 to 13 days of gestation. RBP and TTR mRNAs were both observed in the visceral endoderm as early as at 7 days of gestation. The transcripts were not expressed in the parietal endoderm. At 9 days of gestation, TTR mRNA was strongly detected in the visceral extraembryonic endoderm and in the foregut endoderm, whereas RBP mRNA was detected only in the visceral extraembryonic endoderm. From the 10th day to the 13th day of gestation, both transcripts were increasingly expressed in the visceral yolk sac endoderm and in the developing fetal liver, and gradually decreased in the foregut. At 11 days of gestation, TTR mRNA was first detected in the tela choroidea of the fourth ventricle in the brain; and at 13 days, the TTR mRNA was strongly evident in the developing choroid plexus of the fourth and lateral ventricles. These in situ hybridization studies with embryos at different developmental stages show that RBP and TTR mRNAs are transcribed quite early during embryogenesis. The protein products of these transcripts may play important roles in vitamin A and thyroid hormone metabolism, and in the functions that these regulatory compounds play, in the developing rat embryo.

Animals↗

Ultrasound monitoring of fetal movements. A method for assessing fetal development?

Human fetal movements in utero have been analysed by conventional compound B-scan ultrasound techniques. With practice, movements of specific parts of the fetus can be readily recognised, and even as early as the 6th week after conception gross movements of the whole embryo are visible. Monitoring of fetal movements as an indication of normal or abnormal fetal development appears to have been neglected, and future studies in this important area of antenatal paediatrics are suggested.

Arm↗

Phospholipid molecular species composition of developing fetal guinea pig brain.

Adequate accumulation of polyunsaturated essential fatty acids, in particular docosahexaenoic acid (22:6n-3), into membrane phospholipids is critical for optimal fetal brain development. This process is maximal during the period of rapid neurite outgrowth, neuritogenesis, which precedes the major growth phase, myelination. There is no information about differential changes during gestation to individual brain phospholipid molecular species which contain 22:6n-3. Such details of brain development would be concealed by total fatty acid analysis of isolated phospholipid classes. We have detailed phosphatidylcholine (PC) and phosphatidylethanolamine (PE) molecular species compositions in developing fetal guinea pig brain. Total brain PC concentration increased substantially between 40 and 68 (term) d of gestation, corresponding to myelination, while PE increased in a biphasic manner between 25-35 d, which was coincident with onset of neuritogenesis, and 40-68 d. Fetal brain development was accompanied by complex changes in the concentration of individual phospholipid molecular species. During early gestation (25-40 d) 22:6n-3 was enriched in both PC and PE sn-1 16:0 molecular species. However, between 40 d and term there was no further increase in brain PC 22:6n-3 content, while brain PE was significantly enriched in both PE18:1/22:6 and PE18:0/22:6. We hypothesize that accumulation of 22:6n-3 into sn-1 18:1 and 18:0 species represents establishment of a 22:6n-3-containing membrane PE pool which may be turned over more slowly than sn-1 16:0 species. Identification of specific changes in membrane phospholipids which are associated with defined events in brain development may provide a basis for assigning functional roles to individual molecular species.

Animals↗

Synthesis of platelet activating factor by cholinephosphotransferase in developing fetal rabbit lung.

Developing fetal lung is a possible source of the platelet activating factor (PAF, 1-alkyl-2-acetyl-sn-glycero-3-phosphocholine) present in amniotic fluid of women in labor. We have assayed the microsomal activities of a specific enzyme for the de novo synthesis of PAF in developing fetal and neonatal rabbit lung, 1-alkyl-2-acetyl-glycerol-dependent dithiothreitol-insensitive cholinephosphotransferase. The specific activity of this enzyme increased from 0.92 to 3.60 nmol X min-1 X mg-1 protein between day 21 and day 31 of gestation. In constrast, during this same period the activity of the PAF-biosynthetic cholinephosphotransferase in developing rabbit kidney did not change significantly. The specific activity of the diacyl-glycerol-dependent, dithiothreitol-sensitive cholinephosphotransferase that catalyzes the final step in phosphatidylcholine biosynthesis was not altered during late gestation in either fetal lung or kidney. Previously, increased amounts of pulmonary PAF had been found during the latter stages of gestation (Hoffman, Truong and Johnston (1986) Biochim. Biophys. Acta 879, 88-96) and may be attributed to increased activity of the PAF biosynthetic enzymes found in this investigation. This elevated level of PAF in fetal lung may serve to facilitate breakdown of glycogen that provides, in part, the carbon and energy source for surfactant biosynthesis. In addition, PAF may be secreted in association with surfactant into amniotic fluid in which it may interact with amnion tissue and subsequently participate in the events associated with the initiation of parturition.

Acetyl-CoA C-Acetyltransferase↗

Adrenocorticotrophin and luteinizing hormone responses to N-methyl-D-aspartate during fetal development in sheep.

The hypothalamic mechanisms which underlie the development of the fetal neuroendocrine system are unclear. However, in adult animals neuroactive amino-acids, particularly those acting at the N-methyl-D-aspartate (NMDA) receptor, have been shown to be important transmitters involved in the neuroendocrine regulation of the anterior pituitary gland. In this study we have investigated the potential role of NMDA in the neuroendocrine regulation of fetal pituitary function, by examining the ontogeny of LH and ACTH responses to NMDA during fetal development in sheep. Catheterized fetal sheep were injected with 3 intravenous doses of NMDA (1, 2 and 4 mg/kg; estimated fetal body weight) and saline vehicle on consecutive days between days 110-115, 120-125 and 135-140 gestation (term = 145 days). At each gestational age fetuses also received a pituitary challenge test consisting of CRF (0.4 mu g/kg), AVP (80 ng/kg) and GnRH (125 ng/kg). NMDA caused a significant dose-related increase in ACTH at day 120-125 (P<0.02) and day 135-140 gestation (P<0.001). NMDA had no effect on ACTH secretion at day 110-115. The ACTH response to the highest does of NMDA (4 mg/kg) increased with advancing gestational age, in contrast to the ACTH response to CRF+AVP which was the same at all ages. NMDA caused a significant increase in LH secretion at all gestational ages with the greatest response observed at the latest gestational age studied (day 135-140). A similar increase in LH response to the GnRH challenge was observed late in gestation. Pretreatment of fetuses with the competitive NMDA antagonist CGP37849 (1 mg/kg) abolished the LH and ACTH responses to an intravenous injection of NMDA (4 mg/kg) given 5 min later. These data show that activation of NMDA receptors during fetal development elicits the secretion of ACTH and LH and demonstrate that NMDA receptors are functionally coupled to the neuroendocrine pathways regulating these two hormones during fetal life. The dramatic increase in ACTH response to NMDA during the final days before birth occurs at a time of high adrenocortical activity and suggests an important functional role for these receptors at this time.

2-Amino-5-phosphonovalerate↗

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↗

Anomalies of fetal development in GK rats.

Fetal development was investigated in Goto-Kakizaki (GK) rats. Between days 15 and 20 after identification of a positive sperm plug, the GK rats gained less weight than control animals. The number of conceptuses in each litter was not significantly different in control and GK rats. The incidence of abortive fetal development, however, averaged 39.7% +/- 9.1% in GK rats, compared with only 5.6% +/- 0.2% in control animals. The placental weight in living fetuses was slightly lower in GK rats than control rats. The crown-rump length was identical in the fetuses of control and diabetic mothers. The number of ossification points in the lumbosacral spine, pelvic girdle and anterior and posterior limbs was significantly lower in fetuses of GK rats than control animals. These anomalies could not be blamed on a lower plasma insulin concentration in GK than control animals, whether before or during (days 15-20) pregnancy. Moreover, in both control and GK rats, the insulinogenic index was raised during pregnancy. These findings indicate that GK rats represent a new model for the study of diabetes-related fetal anomalies, their pathogenesis and prevention.

Animals↗

Understanding cellular disruptions during early embryo development that perturb viability and fetal development.

An inability to regulate ionic and metabolic homeostasis is related to a reduction in the developmental capacity of the embryo. The early embryo soon after fertilisation and up until compaction appears to have a reduced capacity to regulate its homeostasis. The reduced ability to regulate homeostasis, such as intracellular pH and calcium levels, by the precompaction-stage embryo appears to impact on the ability to regulate mitochondrial function and maintain adequate levels of energy production. This reduction in ATP production causes a cascade of events leading to disrupted cellular function and, perhaps ultimately, disrupted epigenetic regulation and aberrant placental and fetal development. In contrast, after compaction the embryo takes on a more somatic cell-like physiology and is better able to regulate its physiology and therefore appears less vulnerable to stress. Therefore, for human IVF it would seem important for the establishment of healthy pregnancies that the embryos are maintained in systems that are designed to minimise homeostatic stress, particularly for the cleavage-stage embryos, as exposure to stress is likely to culminate in impaired embryo function.

Adenosine Triphosphate↗

Human macrophage metalloelastase (MMP-12) expression is induced in chondrocytes during fetal development and malignant transformation.

Fetal development and tumor progression both require a complex system of extracellular matrix (ECM) synthesis and breakdown, which is mediated by, for instance, the matrix metalloproteinases (MMPs). Human metalloelastase (MMP-12) is an MMP, the expression of which has so far been documented in macrophages associated with atherosclerosis, wound repair, and certain cancers. In this study we first examined the expression of MMP-12 during human fetal development. By in situ hybridization MMP-12 transcripts were detected in chondrocytes of hypertrophic cartilage in vertebrae of the spinal column, in ribs, and in extremities undergoing ossification, beginning at the gestational age of 8 weeks. Also, periosteal cells expressed MMP-12 at 11 weeks. No expression of MMP-12 mRNA could be noted in other fetal tissues, including the skin, lungs, intestine, kidney, and liver. Expression of MMP-12 mRNA could not be detected in adult normal cartilage or osteosarcomas, but in chondrosarcomas both macrophages (8 of 19 samples) (identified by CD68 immunostaining) and chondrosarcoma cells (8 of 19) were positive. MMP-12 was also demonstrated in the tumors by western blotting and it was expressed in the same regions as MMP-13 mRNA. By immunostaining, MMP-12 mRNA colocalized with the protein in both fetal and chondrosarcoma specimens. Unlike basic fibroblast growth factor (bFGF) and transforming growth factor-beta (TGF-beta), tumor necrosis factor-alpha (TNF-alpha) induced MMP-12 mRNA production in chondrosarcoma-derived HTB-94 cells. Our results suggest that MMP-12 plays an important role in ECM remodeling during fetal bone development and is induced when chondrocytes undergo malignant transformation.

Bone Neoplasms↗

Follicular development, ovulation, fertilization and fetal development in tenrecs (Tenrec ecaudatus).

Reproduction in female tenrecs was studied on Praslin Island (4 degrees 20'S, 55 degrees 45'E) in the Seychelles from November 1977 to September 1980. Dissection and histological examination of reproductive tracts revealed that each ovary is encapsulated in a bursa. Polyovular follicles were abundant and contained up to 5 oocytes. The occurrence of more implanted fetuses than corpora lutea (CL) indicated successful polyovuly in 25% of pregnancies. No antrum was observed during follicular growth at oestrus, and follicles were relatively small at maturation, reaching a maximum diameter of 570 microns. Ovulation involves a gentle extrusion of the granulosa and ovum into the periovarian space. Spermatozoa were abundant in the female reproductive tract including the periovarian space at oestrus. Spermatozoa penetrated the loosely-packed theca of mature follicles and were present in CL, indicating that intrafollicular fertilization may occur. The relatively small CL formed by eversion, reaching a maximum size after implantation. Fusion of up to four CL occurred in the ovaries of 41% of pregnant females. Polyovulation in the tenrec was confirmed by the occurrence of 10.4 +/- 0.5 (mean +/- s.e.m.) CL and 9.7 +/- 0.5 (mean +/- s.e.m.) implantations. Resorption occurred in 74% of 19 dissectable pregnancies, occurring in both uterine horns in 26% of pregnancies. Coefficients of variation in weights of fetuses in each litter were 10.0-26.8%, with the largest value corresponding to 273% variation in fetus weights. Litter size in the Seychelles tenrecs was smaller than in the same species in Madagascar (17.1 +/- 1.9 implantations) where the tenrec is native.

Animals↗