Ossification of the cartilage skeleton of the extremities of human embryos

Certain sequence in appearance of ossification points has been stated in the cartilage models of the superior and inferior extremities of the human embryos at the end of the embryonal and the beginning of the fetal periods of development. The change in the size (length) of the ossification points in anlages of the long tubular bones during the successive stages of embryogenesis is of linear character and can be described by means of the equation y = ax + b, where y--age of the embryo (days), x--length of the osseous points. Coefficients a and b are calculated for estimation the age of the embryos according to the length of the osseous points in the anlages of the brachial, femoral and radial bones.     

Types of neural cells in the spinal ganglia of human embryos and early fetuses

Spinal ganglial of human embryos and fetuses ranging in C.-R. length from 15 to 74 mm and in age from 6 1/2 to 11 postovulatory weeks were studied by light and electron microscopy. A sequence of events in differentiation and maturation enabled five types of cells to be distinguished: 1. apolar, undifferentiated neuroblasts are the main cells at 6 1/2 to 7 1/2 weeks; 2. early bipolar neuroblasts (strictly speaking, types 2 to 5 are immature neurons) predominate at the end of the embryonic period proper (8 postovulatory weeks); 3. intermediate bipolar neuroblasts are characteristic of the early fetal period; 4. late bipolar neuroblasts, in which two proceses arise separately from one pole of the cell, appear at about 10 postovulatory weeks; 5. unipolar neuroblasts are found within another week and, by that time, cells of types 1 and 2 are no longer present.     

Adaptive characteristics of growth of human embryos and fetuses during the intrauterine life]

Under observations was the dynamics of growth of human embryos and fetuses in embryogenesis which is directly dependent on the month of the intrauterine life. Coefficients of correlation and regression were calculated and with their help the average growth of the general length of fetuses in each month of prenancy was determined. The enlargement of the fetus length was found to be more intensive in the embryonic and prefetal periods.     

Development of the skeleton in Japanese quail embryos

In the research fields of experimental embryology, teratological testing, and developmental engineering in avian species, a knowledge of normal embryonic development is necessary so that research may be performed efficiently and precisely. A series of normal stages based on external appearance has been established in both chicken and quail embryos. Those based on skeletal features, however, have not been elucidated. The present study newly established a series of normal stages for the development of the Japanese quail embryo skeleton. This series is composed of 15 stages determined by observing the timing of chondrification and calcification of the skeleton every 24 h, from 3 to 17 days of incubation. Cartilage and ossified bones were stained blue and red with Alcian blue 8GX and alizarin red S, respectively. These skeletogenous stages of the Japanese quail embryo will be useful as a normal control not only in studies of experimental embryology, teratological testing, and developmental engineering, but also in the analysis of mutant embryos with skeletal abnormalities.     

UVB radiation prevents skeleton growth and stimulates the expression of stress markers in sea urchin embryos

Ozone depletion results in an increased flux of biologically damaging radiations reaching the earth. Although ultraviolet (UV) penetration is attenuated by the seawater, harmful effects can be still observed at low depths where sea urchin embryos are living. We have used Paracentrotus lividus embryos to study the impacts of UV radiation on their development. Blastula cultures were exposed to different doses of UVB (312 nm) radiations and the resulting endpoint effects were evaluated in terms of embryonic morphological abnormalities, variations in specific gene expression, and changes in the levels of stress proteins. We found that embryos were moderately sensitive to 50 J/m² UVB radiation; an increase in the number of developmentally delayed and malformed embryos was detected when increasing doses, up to 1000 J/m², were used. Major developmental defects, observed 24 and 48 h after exposure, consisted in the failure of skeleton elongation and patterning. Accordingly, we found a reduction in the number of primary mesenchyme cells that expressed Pl-SM30, a gene coding for one of the specific matrix proteins of the skeleton. The morphological effects observed 1, 24, and 48 h after exposure were correlated with a dose-dependent increase in the level and in the activation of two recognized stress markers, namely hsp70 and p38 MAPk, respectively, consistent with their role in mediating cellular response to stress and suggesting a function in embryo survival.     

Early ontogeny of the central benzodiazepine receptor in human embryos and fetuses

The early ontogeny of the central benzodiazepine receptor (BZR) was investigated in human embryos and fetuses between 7 and 26 weeks of gestation. Brain tissue was gained from terminated pregnancies or spontaneous abortions. Binding studies, which were performed with 3H-flunitrazepam (FNZ), revealed that specific benzodiazepine binding is already detectable at an embryonal age of 7 weeks post conceptionem. Binding at this early stage can be displaced potently by clonazepam and the inverse agonist beta-CCE. Additionally, 3H-FNZ binding is enhanced by GABA. Thus, benzodiazepine binding is of the central type. Receptor density increases steeply in whole brain between weeks 8 and 11 of gestation. In frontal cortex receptor density increases gradually between weeks 12 and 26 of gestation. No specific fetal disease entity (including trisomy 21) was consistently associated with exceptionally high or low Bmax-values.     

IGF-2 stimulates growth and metabolism of early mouse embryos.

Recent reports indicate that the insulin gene family plays a significant role in early development. Both insulin and IGF-1 stimulate growth and metabolism in preimplantation mouse embryos, however, little is known of the physiological effects of IGF-2. In this study, addition of IGF-2 to defined culture medium for the culture of 2-cell embryos stimulated blastocyst formation by 15%, ICM mitogenesis by 37%, and protein synthesis by 35%. EC50s of 12-63 pM IGF-2 for these responses were in the range for mediation by IGF-2 receptors. These results coupled with the previously demonstrated presence and expression of the IGF-2 receptor from the 2-cell stage supports a role for this third member of the insulin gene family in early development.     

Development of the notochord in normal and malformed human embryos and fetuses.

In view of its possible involvement in early embryogenesis and teratogenesis, the developmental characteristics of the human notochord were studied by light and electron microscopy and immunohistochemistry on 20 human conceptuses (5th-22nd week). At the earliest embryonic stages examined, the notochord is closely related to both the pharyngeal endoderm and the neuroectoderm of the posterior (tail) end of the neural tube. In both regions the interspace is bridged by slender cytoplasmic processes, lined with basal lamina and filled with amorphous extracellular material containing collagen types III and IV and laminin. The notochordal cells express cytokeratin brightly and vimentin weakly. As embryonic age progresses, the notochord gradually separates from the epithelia, becomes the axis of developing spinal column and undergoes progressive cell degeneration and rearrangement within the vertebral bodies. This is associated with extensive production of extracellular material and the first appearance of fibronectin. Intracellularly, the expression of vimentin gradually increases, while that of cytokeratin slightly weakens. Changes in the notochord parallel other developmental events in axial organs. In anencephalic fetuses the course of the notochord is irregular and partly interrupted with segments outside the basichondrocranium in specimens with craniorachischisis.     

Immunohistochemical localization of entactin and laminin in mouse embryos and fetuses

The temporal appearance and the anatomic distribution of entactin in the developing mouse embryo was studied by light and electron microscopic immunohistochemistry and compared with the appearance and the distribution of laminin. Immunohistochemically detectable entactin first appeared in the hatched blastocysts, in contrast to laminin which became apparent as early as the 8-cell stage of development. However, beginning with the appearance of entactin in the hatched blastocyst, antibodies to these two basement membrane glycoproteins co-localized in all extraembryonic matrices. These findings indicate that the synthesis of entactin and laminin is asynchronous in the early stages of mouse embryonic development but becomes synchronized in the postimplantational stages of development.     

Plakoglobin is required for maintenance of the cortical actin skeleton in early Xenopus embryos and for cdc42-mediated wound healing

Early Xenopus embryos are large, and during the egg to gastrula stages, when there is little extracellular matrix, the cytoskeletons of the individual blastomeres are thought to maintain their spherical architecture and provide scaffolding for the cellular movements of gastrulation. We showed previously that depletion of plakoglobin protein during the egg to gastrula stages caused collapse of embryonic architecture. Here, we show that this is due to loss of the cortical actin skeleton after depletion of plakoglobin, whereas the microtubule and cytokeratin skeletons are still present. As a functional assay for the actin skeleton, we show that wound healing, an actin-based behavior in embryos, is also abrogated by plakoglobin depletion. Both wound healing and the amount of cortical actin are enhanced by overexpression of plakoglobin. To begin to identify links between plakoglobin and the cortical actin polymerization machinery, we show here that the Rho family GTPase cdc42, is required for wound healing in the Xenopus blastula. Myc-tagged cdc42 colocalizes with actin in purse-strings surrounding wounds. Overexpression of cdc42 dramatically enhances wound healing, whereas depletion of maternal cdc42 mRNA blocks it. In combinatorial experiments we show that cdc42 cannot rescue the effects of plakoglobin depletion, showing that plakoglobin is required for cdc42-mediated cortical actin assembly during wound healing. However, plakoglobin does rescue the effect of cdc42 depletion, suggesting that cdc42 somehow mediates the distribution or function of plakoglobin. Depletion of alpha-catenin does not remove the cortical actin skeleton, showing that plakoglobin does not mediate its effect by its known linkage through alpha-catenin to the actin skeleton. We conclude that in Xenopus, the actin skeleton is a major determinant of cell shape and overall architecture in the early embryo, and that plakoglobin plays an essential role in the assembly, maintenance, or organization of this cortical actin.     

Relationship between parameters of early growth in Israeli infants

A sample of 1931 Israeli infants was measured for body weight (WT), length (HT) and head circumference (HC) for approximately 2 years. The Count model with 3 parameters was chosen as the best fitting and most parsimonious function to approximate growth of all 3 studied traits. In the model parameter a relates to birth indices, b--to velocity of growth, and c--to rapid early childhood growth, or acceleration. Assuming a difference in growth patterns in the periods of different length, the whole sample was divided into 3 groups: 1) infants with last measurement around the age of 12 months; 2) infants with last measurement around the age of 18 months, and 3) infants with last measurement around the age of 24 months. The individuals measured up to 12 months were presented in all three groups. 27 curve fitting parameters, corresponding to 3 different follow-up intervals for WT, HT and HC were computed for each individual. A high correlation was detected between the a parameters regardless of time interval for 3 measured traits. A negative correlation was found between b and c parameters within the same time interval. A consistent positive correlation was indicated between a and b parameters, especially for body length and head circumference. A principal component analysis extracted five independent factors explaining 88.1% of the total variance. Three first factors retained parameters b and c, describing growth rate and pattern of each trait separately, namely, F1 was responsible for head circumference, F2 was a body length factor, F3 was a body weight factor. F4 extracted all birth indices, observed (HC0, HT0 and WT0) and expected (parameters a). The composition of principal factors allowed us to assume that there might be a strong involvement of a pleiotropic genetic source in determination of birth size traits and an independent genetic source controlling the pattern of growth for each trait separately.