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Chromosomal arrangement of the duck alpha-globin genes and primary structure of the embryonic alpha-globin gene pi.

A recombinant lambda Charon 4A bacteriophage, D alpha G-1, carrying the genes coding for the duck embryonic (pi') and adult (alpha A, alpha D) alpha-like globins was isolated from a previously constructed duck DNA recombinant library. The three globin genes are transcribed from the same DNA strand and are arranged in the order of their expression during development: 5'-pi'-alpha D-alpha A-3'. We have determined the complete nucleotide sequence of the duck pi'-globin gene, including the flanking regions. Due to the unusual length of intron 1 (963 bp) and intron 2 (568 bp) the 2167-bp duck pi'-globin gene is by far the largest among all known mammalian or avian alpha- and beta-globin genes. For instance, the duck pi'-globin gene introns are almost twice as long as those of the chicken pi'-globin genes. A surprisingly high degree of nucleotide sequence homology (88%) has been found for the 5' flanking region (positions -1 to -223) of the duck and chicken pi'-globin gene.

Amino Acid Sequence↗

Structure and early embryonic expression of the zebrafish engrailed-2 gene.

The Drosophila homeobox gene engrailed (en) is needed for correct embryonic development, and related sequences are active during vertebrate embryogenesis. Here we report the protein coding sequence and embryonic expression pattern of the zebrafish engrailed-2 gene (eng-2) which is directly homologous to En-2 in mice and Xenopus. The predicted zebrafish Eng-2 protein shares 65% overall identity to its Xenopus counterpart. In addition to the highly conserved homeodomain region, sequence conservation is present within three short stretches in the N-terminal region. The embryonic expression of the eng-2 gene was analysed by in situ hybridization to whole-mount embryos and tissue sections. Transcripts are first detected in two lateral bands at the 10-h stage, when epiboly is completed. Within the next 2 h of development, these two bands migrate and fuse at the midline. By the time the neural keel becomes visible (11-12 h), a transverse stripe of eng-2 expressing cells is seen at the presumptive midbrain-hindbrain boundary. Later this stripe becomes significantly compressed along the AP axis, and in 24-h embryos eng-2 transcripts are detected mainly in the posterior midbrain. In the hindbrain, eng-2 expression seems restricted to the primordium of the cerebellum. A second site of activity was observed in each somite where specific myotomal cells, the muscle pioneers, express eng-2. Our observations are discussed in relation to early regionalization of the central nervous system (CNS) and the generation of morphological borders.

Amino Acid Sequence↗

Redox control of angiogenic factors and CD31-positive vessel-like structures in mouse embryonic stem cells after direct current electrical field stimulation.

The molecular mechanisms driving angiogenesis in tissues derived from embryonic stem (ES) cells are currently unknown. Herein we investigated the effects of direct current (DC) electrical field treatment on endothelial cell differentiation and angiogenesis of mouse ES cells. Treatment of ES cell-derived embryoid bodies with field strengths ranging from 250 V/m to 750 V/m, applied for 60 s, dose-dependently increased the capillary area staining positive for the endothelial-specific marker platelet endothelial cell adhesion molecule-1 (PECAM-1), indicating stimulation of endothelial cell differentiation and angiogenesis. Consequently, increased expression of hypoxia-inducible factor-1alpha (HIF-1alpha) and vascular endothelial growth factor (VEGF) within 24 h was observed. Electric field treatment raised reactive oxygen species (ROS) generation for at least 48 h, which was blunted by NADPH-oxidase inhibitors diphenylen iodonium chloride (DPI) as well as 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF), and increased the expression of NADPH-oxidase subunits p22-phox, p47-phox, p67-phox, and gp91-phox within 24 h. Electrical field treatment resulted in activation of extracellular regulated kinase 1,2 (ERK1,2), p38, as well as c-Jun NH2-terminal kinase (JNK). Pretreatment with the JNK inhibitor SP600125 resulted in a significant decrease in capillary areas under control conditions as well as under conditions of electrical field treatment, whereas the p38 inhibitor SB203580 was without effects. By contrast, the ERK1,2 antagonist UO126 inhibited electrical field-induced angiogenesis, whereas angiogenesis under control conditions was unimpaired. The increase in capillary areas and VEGF expression as well as activation of JNK and ERK1,2 was significantly inhibited in the presence of the free radical scavenger vitamin E underscoring the role of ROS in electrical field-induced angiogenesis of ES cells.

Angiogenesis Inducing Agents↗

Placental fine structure after experimental embryonic death in cows.

Early embryonic death was experimentally induced by amnion rupture in pregnant heifers. The subsequent morphological alterations in the placenta were studied with light and electron microscopy, histochemical tests and electron microscope microanalysis. The material from the treated heifers differed from that inuntreated cows in the following aspects: the fetal-maternal junction was firm; many endometrial epithelial cells were desquamated and the remaining epithelium showed vacuolization or depletion of organelles; the trophoblast remained at the preattachment stage in its marked phagocytic activity and accumulation of lipid and glycogen, and many of the organelles were degenerated; and the mesenchyme contained extracellular deposits of iron and calcium. It is believed that the changes observed were caused by interruption of the fetal circulation followed by an accumulation of metabolic products.

Animals↗

A fine-structural study of embryonic and larval development in the gymnoblastic hydroid Pennaria tiarella.

1. The pregastrulation blastomers contain electron-dense granules which become localized after gastrulation in the apices of the developing epithelio-muscle cells and persist throughout larval development. The cytoplasm of the blastomeres is organized into anucleate, membrane-delimited lobules. The lobules, which persist until six hours of development, come to contain a single, peripherally located cisterna of granular endoplasmic reticulum. Microvilli are present at the earliest stages examined and persist throughout development. Cilia are first detected at four hours. 2. Gastrulation, marked by the appearance of the mesoglea, occurs between six and eight hours of development. Basal foot processes of epithelio-muscle cells are detected by eight hours, but myonemes cannot be detected until later in development. 3. Immediately following gastrulation, mucous cells begin their differentiation from dividing cells located near the apex of the ectoderm. During their differentiation, the cells elongate toward the mesoglea. 4. By 16 hours post-fertilization, a third cell type can be detected in the ectoderm. The cell, which contains no granules, has an unusual cytoplasmic organization in which fused membranes divide the cytoplasm into parallel compartments containing a single cisterna of granular endoplasmic reticulum. 5. The findings of the present study are correlated with those of previous studies of development in Pennaria and other hydroids. The possible functional roles of the Type I granules, the cytoplasmic lobules, and the nongranular cell are discussed.

Animals↗

Fine structure of chick embryonic parathyroid glands cultured on media with different concentrations of calcium.

An electron microscopical study of parathyroid glands from 13-day old chick embryos cultured for 2 days on media with different concentrations of calcium was conducted. The same as in non-cultured embryonic glands, the cells in all cultures contained a moderate amount of rough endoplasmic reticulum and a large Golgi complex with many cisternae, vacuoles, vesicles, coated vesicles and small prosecretory granules. In addition, large secretory granules, which are very rare in the non-cultured controls appeared frequently in the cultures and were especially numerous in the glands cultured on high-calcium medium. The fact that the amount of secretory granules varied according to the concentration of calcium in the medium is interpreted as indicating that the rate of parathyroid hormone secretion in the embryo is already responsive to variations in the concentration of calcium. To the extent that in vitro results may be accepted as representative of what happens in vivo these results support the idea that the embryonic gland may be controlled by variations in the calcemia as the adult gland does.

Animals↗

Analysis of the proteoglycans synthesized by corneal explants from embryonic chicken. II. Structural characterization of the keratan sulfate and dermatan sulfate proteoglycans from corneal stroma.

Radioisotopically labeled proteoglycans were isolated from a 4 M guanidine HCl, 2% Triton X-100 extract of corneal stroma from day 18 chicken embryos by anion-exchange chromatography. Two predominant proteoglycans in the sample were separated by octyl-Sepharose chromatography using a gradient elution of detergent in 4 M guanidine HCl. One proteoglycan had an overall mass of approximately 125 kDa, a single dermatan sulfate chain (approximately 85-90% chondroitin 4-sulfate, low iduronate content) of approximately 65 kDa, and a core protein after chondroitinase ABC digestion of approximately 45 kDa which also contained one to three N-linked oligosaccharides and one O-linked oligosaccharide. The other proteoglycan had an overall size of approximately 100 kDa, two to three keratan sulfate chains of approximately 15 kDa each, and a core protein following keratanase digestion of approximately 51 kDa which included two to three N-linked but no O-linked oligosaccharides. A larger size, a greater overall hydrophobicity (as measured by its interaction with octyl-Sepharose) and an absence of O-linked oligosaccharides argue that this core protein is a distinct gene product from the core protein of the dermatan sulfate proteoglycan.

Animals↗