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polyhomeotic: a gene required for the embryonic development of axon pathways in the central nervous system of Drosophila.

Hypomorphic alleles of the locus polyhomeotic (ph) produce multiple, homeotic-like transformations in adult flies that mimic dominant mutations in the Antennapedia and Bithorax complexes. Analysis of null alleles of ph has revealed a complex, embryonically lethal phenotype that includes cell death of the ventral epidermis and abnormalities in the patterns of expression of homeotic and segmentation genes. There is also a dramatic alteration in the pattern of axon pathways in the central nervous system, such that the wild-type array of segmentally repeated commissures and connectives is replaced by bundles of axons confined to the hemiganglia of origin. It is possible that this axonal phenotype is the result of loss of neuronal identity caused by abnormal homeotic and segmentation gene expression.

Animals↗

Platelet-derived growth factor receptor alpha (pdgfr-alpha) gene in zebrafish embryonic development.

Here we present the cloning of a full-length zebrafish pdgfr-alpha cDNA as well as the expression of this gene during zebrafish embryogenesis. We show that zebrafish pdgfr-alpha mRNA is present at high levels in the fertilized egg as well as in all embryonic cells up to the end of gastrulation. Spatially restricted expression of the gene started after the onset of segmentation and is mainly localized in premigratory neural crest cells, the placodes, the anterior paraxial cells of somites and the adaxial cells of the tailbud. Transient expression of this gene was also detected in the early Kupffer's vesicle, a teleost-specific structure. Expression of the zebrafish pdgfr-alpha is both conserved as well as diverged comparing to that of other vertebrate species.

Animals↗

The metabolism of astaxanthin during the embryonic development of the crayfish Astacus leptodactylus Eschscholtz (Crustacea, Astacidea).

The developing eggs of the crayfish Astacus leptodactylus are found to contain important carotenoid amounts. Free astaxanthin, mainly associated with protein and lutein represent the main pigments occurring in the yolk at the end of the embryonic period. Esterification processes affecting astaxanthin appear concomitantly with the resorption of vitellin reserves, the activation of the digestive gland and the rapid decrease of lutein amounts. Esterified astaxanthin contents increase significantly within the interval between the juvenile stages I and II. Nevertheless, free astaxanthin levels appear insufficient to balance the amounts of the esterified form, scored at the end of the endotrophic period.

Animals↗

Delayed embryonic development of mouse masseter muscle correlates with delayed MyoD family expression.

While the masseter muscle is known to have several unique developmental characteristics as compared with other skeletal muscles, little is known about its myogenesis. Thus, we examined the expression of myogenic marker and of myoD family gene mRNA from embryonic day (E) 11 to birth. The obtained results were compared with our earlier results of the mouse tongue muscle, which is also involved in oral functions. The mRNA quantities were determined by means of the reverse-transcription and competitive-polymerase chain-reaction techniques. The expression of myogenic marker mRNA indicated that differentiation and maturation in the masseter began at E13 as in the tongue, and were not yet completed at birth, although they were completed in the tongue. The expression of myoD, myogenin, and myf5 mRNA peaked later in the masseter (E17) than in the tongue (E13). The expression of MRF4 mRNA began later in the masseter (E15) than in the tongue (E13). These results suggest that the delayed expression of the myoD family genes in the masseter correlates with delayed differentiation and maturation, probably due to the later functional requirements of the masseter than of the tongue.

Animals↗

Expression of the mouse FGF-1 and FGF-1.A mRNAs during embryonic development and in the aging heart.

The mouse Fgf-1 gene contains at least four upstream promoters that are alternatively spliced to the first protein coding exon, giving rise to different Fgf-1 mRNA variants (1.A, 1.B, 1.C, and 1.G), each expressed in a tissue specific manner. Only the Fgf-1.A promoter contains TATA and CAAT consensus sequences, and its corresponding mRNA is mainly expressed in the mouse heart. In situ hybridization with an Fgf-1.A-specific cRNA probe showed the 1.A message to be very low in embryonic heart, tongue, spinal cord and smooth muscle of the small intestine. Moderate 1.A mRNA levels were revealed in head mesenchyme, paraxial mesoderm, in the embryonic inferior cunniculus, hyppocampus, and thymus. These results suggest a limited role of the Fgf-1.A mRNA variant during development.

Aging↗

Sodium, phosphorus, sulphur, chlorine and potassium shifts in rat brain during embryonic development.

Concentrations of sodium (Na), phosphorus (P), sulphur (S), chlorine (Cl) and potassium (K) and their variations during brain development were measured in freeze-dried thick sections from rat brain (16-20 micron). Sprague-Dawley rats were bred and the day of finding vaginal spermatozoa was considered as day zero of pregnancy. On days 12E, 13E, 14E, 16E, 19E, 21E (embryonic) and postnatal day one whole embryos or fetal heads were rapidly frozen in liquid Freon 22 cooled with liquid nitrogen (-180 degrees C), sectioned in a cryostat (-20 to -40 degrees C), and processed for X-ray microanalysis on pure carbon plates. Concentrations of Na and Cl differed in the cells of the cerebral cortex, ependyma, choroid plexus and cerebral spinal fluid (CSF). During cerebral development, Na and Cl concentrations appeared to be correlated, while K was more related to P. S was low and unchanged in all compartments during development and was thus considered as an internal control. K was inversely related to Na and Cl fluctuations within the choroid plexus epithelia. Sharp phase changes of elemental composition appeared in all tissues at specific growth stages, e.g. days 14E and 19E. These results demonstrate rhythmic changes in the inorganic components of developing rat brain cells and fluid environment presumably reflecting physiological fluctuations and cell cycle phenomena. Such changes may also be related directly or indirectly to known 'growth phase changes' in the developing rat.

Animals↗

Lats2/Kpm is required for embryonic development, proliferation control and genomic integrity.

The Drosophila melanogaster warts/lats tumour suppressor has two mammalian counterparts LATS1/Warts-1 and LATS2/Kpm. Here, we show that mammalian Lats orthologues exhibit distinct expression profiles according to germ cell layer origin. Lats2(-/-) embryos show overgrowth in restricted tissues of mesodermal lineage; however, lethality ultimately ensues on or before embryonic day 12.5 preceded by defective proliferation. Lats2(-/-) mouse embryonic fibroblasts (MEFs) acquire growth advantages and display a profound defect in contact inhibition of growth, yet exhibit defective cytokinesis. Lats2(-/-) embryos and MEFs display centrosome amplification and genomic instability. Lats2 localizes to centrosomes and overexpression of Lats2 suppresses centrosome overduplication induced in wild-type MEFs and reverses centrosome amplification inherent in Lats2(-/-) MEFs. These findings indicate an essential role of Lats2 in the integrity of processes that govern centrosome duplication, maintenance of mitotic fidelity and genomic stability.

Animals↗

The flightless I protein localizes to actin-based structures during embryonic development.

The product of the flightless I gene is predicted to provide a link between molecules of an as yet unidentified signal transduction pathway and the actin cytoskeleton. Previous work has shown that weak and severe mutations of the flightless I locus in Drosophila melanogaster cause disruption in the indirect flight muscles and in embryonic cellularization events, respectively, indicative of a regulatory role for the flightless I protein in cytoskeletal rearrangements. A C-terminal domain within flightless I with significant homology to the gelsolin-like family of actin-binding proteins has been identified, but evidence of a direct interaction between endogenous flightless I and actin remains to be shown. In the present study, chick, mouse and Drosophila melanogaster embryos have been examined and the localization of flightless I investigated in relation to the actin cytoskeleton. It is shown that flightless I localization is coincident with actin-rich regions in parasympathetic neurons harvested from chicks, in mouse blastocysts and in structures associated with cellularization in Drosophila melanogaster.

3T3 Cells↗

Activity of cholinesterases in the Japanese quail embryo. Effects of dichlorphos on the embryonic development.

Cholinesterase activity is detectable in the Japanese quail embryo, in the yolk and subembryonic liquid, but not in the albumen. Obviously, this enzyme is deposited by the hen into the yolk and from there it is transferred to the subembryonic liquid. In contrast, in the embryo the enzyme is synthesized by itself and the amount increases with the age of the embryo. By using BW284c51 1,5-bis-(4-allyldimethylammoniumphenyl)pentan-3-one bromide and ISO-OMPA tetraisoprophylpyrophosphoramide as inhibitors, it was found that the enzyme in the embryo is predominantly acetylcholinesterase (EC 3.1.1.7), whereas that in the yolk and subembryonic liquid is butyrylcholinesterase (EC 3.1.1.8). Both types are inhibited by dichlorphos. However, the embryonic enzyme activity is restored within 8 hr, whereas that in the subembryonic liquid remained inactive at least for 72 hr after inhibition. Enzyme inhibition leads to retardation of the development, to reduced accumulation of glucose and amino acids in the subembryonic liquid and finally to death of the embryo, suggesting that the developmental retardation is due to the restricted supply of glucose and amino acids. Surprisingly, most of the embryos die when the embryonic enzyme activity has again been restored.

Amino Acids↗

Embryonic development of spontaneous motility in chick embryos. Identification of participation of NMDA-mechanism by magnesium ions.

The influence of magnesium ions on the spontaneous motility and its development was studied in chick embryos aged from day 4 to day 19 of incubation. 1-An acute application of MgCl2 (80 mg/kg e.w.) evoked from day 13 of incubation a significant depression of spontaneous motility together with decreased synchronization of embryonic spontaneous movements. 2-The resting spontaneous motility in 17-day-old embryos after chronic continual application of MgCl2 (11.5 +/- 1.21 mg/kg e.w./24 h.) from day 4 to day 12 (Mg2+/ser.C) was reduced and exhibited smaller synchronization of spontaneous movements by the preponderance of movements with moderate amplitude. 3-The chronic application of MgCl2 expressed itself also by the changed reactivity of spontaneous motility to the factors influencing NMDA- and NO-ergic mechanisms. The decrease of NMDA activatory effect, the weakened depressory effect of ketamine, the reduction of periodic depressive effect of arcaine and the reduction of activatory effect of cocktail (NMDA + L-arginine) took place. 4-The investigated effects of Mg2+ confirm the participation of NMDA-ergic mechanism and simultaneously point out the possible role of NO-ergic mechanism in the genesis and the development of spontaneous motility in chick embryos.

Animals↗

Ultrasonographic imaging to monitor early pregnancy and embryonic development in the buffalo (Bubalus bubalis).

Real time B-mode ultrasound was used to detect and monitor the early conceptus, its growth and its anatomical features in 26 buffalo between Days 18 and 62 of gestation. The buffalo were artificially inseminated, and the conceptuses were examined on alternate days beginning on Day 18. The embryonic vesicle and the embryo proper within the vesicle was first visible in 12 of the buffalo on mean Day (+/-SD) 19.00+/-2.1 and Day 19.0+/-1.69, respectively. The heartbeat of the embryo proper could be detected on Day 29.6+/-1.57. The heart rate of 203.8+/-9.0 beats per minute on the first day of detection decreased to 150 beats per minute on Day 62. The allantois, amnion, fore limbs, spinal cord and hind limbs were first identified on Day 30.0+/-1.14, Day 33.4+/-1.64, Day 34.6+/-1.34, Day 35.8+/-2.52 and Day 36.8+/-2.34, respectively. The optic area was first distinguished on Day 38.2+/-2.39. Split hooves, fetal movement, ribs and vertebra were identified on Day 46.0+/-2.64, Day 49.4+/-2.31 and Day 59.8+/-2.39, respectively. The mean length of the embryo proper was 4.2 mm on Day 19 which later increased to 53.6+/-2.1 mm on Day 62.

Journal Article↗

Embryonic development in the pig up to the 64-cell stage, with reference to DNA replication and cell cycle times from the third cleavage division.

Preimplantation cleaving-stage embryos were recovered from Dutch Landrace (DL) and F1 Dutch Landrace x Great Yorkshire (DL x GY) gilts for which the time of insemination and ovulation were known. Embryonic cell counts were performed, usually after brief in vitro culture to estimate DNA synthesis. Special attention was given to the 4-cell stage. Beyond this stage, the mean cell cycle time was 14 hours for DL and 17 hours for F1 gilts. Generally, high indices of DNA synthesis were obtained (more than 60% of nuclei). There was prominent within and between gilt variability with regard to embryonic cell numbers. Gilts are especially heterogeneous with respect to the length of the 4-cell stage. The G 2 M phase of the 4-cell stage takes approximately 3.5 hours. Especially for F1 gilts, the age of the spermatozoa at the moment of ovulation was not related to the rate of cleavage and/or embryonic death. It is postulated that variability in the length of the 4-cell stage is reflected in genetic activation of the embryos at this stage and subsequently influences embryonic survival.

Journal Article↗

Alteration of Ca2+ homeostasis of sea urchin embryos by retinoid CD 367, dual effect on egg cleavage and embryonic development.

The effect of a light stable retinoid (CD 367) was studied on sea urchin embryos. CD 367 did not affect sperm-egg interaction. In a range of concentrations between 10 and 100 microM, CD 367 delayed the first and the second cleavages. When added after fertilization, micromolar amounts of CD 367 delayed hatching and produced embryonic abnormalities in a dose-dependent manner. Mesodermal cells, primary (PMC) and secondary (SMC) mesenchyme cells migration was particularly disturbed, leading to exogastrulations and calcified spicules malformations. Concentrations of CD 367 higher than 8 microM were embryolethal. Micromolar amount of CD 367 increased plasmalemma Ca2+ permeability of fertilized eggs but not of unfertilized eggs. CD 367 inhibited ATP-dependent intracellular sequestration of Ca2+ in a range of concentrations similar to those affecting egg cleavage and embryonic structures. Since we were unable to detect nuclear receptors for CD 367 in sea urchin eggs and ovocytes, these effects probably are not related to interaction of the retinoid with members of the RAR family, to which CD 367 has a high affinity, but rather to its toxicity by the means of some unknown mechanisms.

Animals↗

Recent advances in our understanding of the temporal control of early embryonic development in amphibians.

Recent studies on temporal control of early amphibian development are reviewed. It is becoming clear that the development of an embryo is not timed by a single clock set in motion at fertilization, instead each developmental event seems to be timed by its own clock-like mechanism. The timing of developmental events is rigidly determined within embryonic cells, and usually can not be altered experimentally. One exception, however, is the timing of midblastula transition in amphibian embryos; recent studies have shown that its timing is regulated by the nucleocytoplasmic ratio. Several developmental events, particularly those associated with transcriptional activities, require DNA replication prior to their occurrence, suggesting an intimate relationship between DNA replication cycles and their onset. On the other hand, there are many other developmental events where timing is not controlled by the number of cell divisions, DNA replication cycles, or the nucleocytoplasmic ratio. Cytoplasmic machinery with autonomous oscillatory properties is thought to be involved in the timing of these events.

Ambystoma↗

The chlorinated AHR ligand 3,3',4,4',5-pentachlorobiphenyl (PCB126) promotes reactive oxygen species (ROS) production during embryonic development in the killifish (Fundulus heteroclitus).

Exposure to dioxin-like chemicals that activate the aryl hydrocarbon receptor (AHR) can result in increased cellular and tissue production of reactive oxygen species (ROS). Little is known of these effects during early fish development. We used the fish model, Fundulus heteroclitus, to determine if the AHR ligand and pro-oxidant 3,3',4,4',5-pentachlorobiphenyl (PCB126) can increase ROS production during killifish development, and to test a novel method for measuring ROS non-invasively in a living organism. The superoxide-sensitive fluorescent dye, dihydroethidium (DHE), was used to detect in ovo ROS production microscopically in developing killifish exposed to PCB126 or vehicle. Both in ovo CYP1A activity (ethoxyresorufin-o-deethylase, EROD) and in ovo ROS were induced by PCB126. In ovo CYP1A activity was inducible by PCB126 concentrations as low as 0.003 nM, with maximal induction occurring at 0.3 nM PCB126. These PCB126 concentrations also significantly increased in ovo ROS production in embryonic liver, ROS being detectable as early as 5 days post-fertilization. These data demonstrate that the pro-oxidant and CYP1A inducer, PCB126, increases both CYP1A activity and ROS production in developing killifish embryos. The superoxide detection assay (SoDA) described in this paper provides a semi-quantitative, easily measured, early indicator of altered ROS production that can be used in conjunction with simultaneous in ovo measurements of CYP1A activity and embryo development to explore functional relationships among biochemical, physiological and developmental responses to AHR ligands.

Animals↗

Expression of Drosophila Ten-a, a dimeric receptor during embryonic development.

Ten-a and Ten-m are the two Drosophila members of the newly discovered Ten-m family of dimeric type II transmembrane proteins. Here, we report complete cDNA cloning and protein expression patterns of Ten-a. The Ten-a protein, a dimeric receptor of about 500 kDa is mainly expressed on axons of the embryonic central nervous system and on muscle attachment sites.

Amino Acid Sequence↗

Tissue specificity of myocardial glycopeptides synthesized during early embryonic development.

[3H] Glucosamine-labeled glycopeptides synthesized by hearts from stage 14-16 chick embryos were separated on DE-52 by ion-exchange chromatography and compared to those synthesized in other parts of the embryo at the same stage. Relatively few molecular species were present in all tissues, and they appeared closely homologous. There were no glycopeptides unique to any embryonic region examined, suggesting that tissue specificity may result from quantitative rather than qualitative differences in glycopeptides.

Animals↗

Isolation and expression of PASK, a serine/threonine kinase, during rat embryonic development, with special emphasis on the pancreas.

We report the isolation and characterization of a serine/threonine kinase expressed during rat pancreas development. This kinase was cloned as part of a general screen using degenerate oligonucleotides to map expression of kinases and receptors during the course of pancreatic development. Sequence analysis showed it to be a member of the ste20-like serine/threonine kinase family. Northern blotting analysis against both fetal and adult tissues showed two transcripts, one of 2 kb and the other of 4 kb. The ratio of transcript expression varied with the tissue. In situ hybridization analysis showed that this gene is expressed in the early gut and pancreatic epithelium. By embryonic Day 15, the transcript is localized to cells that will eventually become exocrine in nature. In situ hybridization analysis also demonstrated high levels of expression in the choroid plexus, the developing myocardium, kidney, CNS, dorsal root ganglia, and testes. In addition, a search of the EST database revealed a related human kinase not previously described.

Amino Acid Sequence↗