Development Genes and Evolution

doi: 10.1007/BF00190230pmid: 28306015

Zivkovic, Danica; Créton, Robbert; Dohmen, René

doi: 10.1007/BF00190231pmid: 28306016

During the first four mitotic division cycles of Lymnaea stagnalis embryos, we have detected cell cycle-dependent changes in the pattern of transcellular ionic currents and membrane-bound Ca2+-stimulated ATPase activity. Ionic currents ranging from 0.05 to 2.50 μA/cm2 have been measured using the vibrating probe technique. Enzyme activity was detected using Ando's cytochemical method (Ando et al. 1981) which reveals Ca2+/Mg2+ ATPase localization at the ultrastructural level, and under high-stringency conditions with respect to calcium availability, it reveals Ca2+-stimulated ATPase. The ionic currents and Ca2+-stimulated ATPase localization have in common that important changes occur during the M-phase of the cell cycles. Minimal outward current at the vegetal pole coincides with metaphase/anaphase. Maximal inward current at the animal pole coincides with the onset of cytokinesis at that pole. Ca2+-stimulated ATPase is absent from one half of the embryo at metaphase/anaphase of the two- and four-cell stage, whereas it is present in all cells during the remaining part of the cell cycle. Since fluctuations of cytosolic free calcium concentrations appear to correlate with both karyokinesis and cytokinesis, we speculate that part of the cyclic pattern of Ca2+-stimulated ATPase localization and of the transcellular ionic currents reflects the elevation of cytosolic free calcium concentration during the M-phase.

Mari-Beffa, Manuel; Celis, José; García-Bellido, Antonio

doi: 10.1007/BF00190232pmid: 28306017

The role of the achaete-scute complex and extramacrochaetae, Notch, Delta, Enhancer of split and Hairless genes in chaeta patterning in Drosophila tergites was studied in genetic mosaics and in mutant combinations. The mutant phenotypes of different alleles of each gene can be ordered in characteristic topographical seriations. These seriations are related to the pattern of proliferation of histoblasts and the time of singularization of sensory organ mother cells from surrounding epidermal cells. Genetic mosaics of lethal alleles show that these genes are fundamentally involved in this singularization and subsequent differentiation. The study of mutant combinations of alleles of these genes reveals specific relationships of epistasis and synergism between them. The results suggest that spatial and temporal variations in achaete-scute complex functional products in cells, modulated by the activity of other genes involved in signal transduction, define the patterned differentiation of sensory organs in tergites.

Schlawny, Axel; Pfannenstiel, Hans-Dieter

doi: 10.1007/BF00190233pmid: 28306018

Blastomeres of two-cell, four-cell, and eight-cell embryos of Hydractinia echinata were injected with horseradish-peroxidase (HRP) or fluorescein isothiocyanate (FITC)-dextran. The fate of the descendants of the injected blastomeres was followed until the planula larva had developed. The results obtained after HRP or FITC-dextran injection were essentially the same. Blastomeres are equivalent up to the four-cell stage, i.e. half-blastomeres produce half of the ectoderm of the planula larva and quarter-blastomeres give rise to one quarter of the larval ectoderm. During normal embryogenesis, the larval anterior-posterior axis corresponds to the animal-vegetal axis of the zygote. Thus, the labelled areas of larvae consisting of the progeny of injected half or quarter blastomeres normally stretch along the larval anterior-posterior axis. Normally, material giving rise to anterior or posterior larval parts, respectively, is separated at the third cleavage. Irrespective of the type of experiment, the progeny of injected blastomeres always contributed to endoderm formation, i.e. in larvae resulting from injected embryos the endoderm was more or less uniformly labelled. Application of vital stains locally to the exterior of zygotes and following these markers through first and second cleavage, produced evidence that in the vast majority of cases, the second cleavage is meridional.

Bidmon, Hans-Jürgen; Granger, Noelle; Stumpf, Walter

doi: 10.1007/BF00190234pmid: 28306019

The presence of c-fos, a marker for cell activation, was investigated in cerebral neurons actively expressing ecdysteroid receptors during larval-pupal development in the tobacco hornworm, Manduca sexta. Colocalization was accomplished by ecdysteroid autoradiography using the tritiated high affinity 20-hydroxyecdysone agonist ponasterone A and immunocytochemistry with an antibody to a peptide sequence which is highly conserved in both human and murine c-fos. Immunoreactivity to a c-fos-like protein(s) was present in nuclei of many neurons of all the developmental stages examined. However, with the exception of the optic lobe, cells expressing nuclear ecdysteroid receptors were more immunoreactive than non-ecdysteroid-binding neurons. These data suggest that ecdysteroid-induced gene activation and translation may involve c-fos expression.

Semik, Danuta; Kilarski, Wincenty

doi: 10.1007/BF00190235pmid: 28306020

We have studied the surface of the animal half of ovulated newt eggs recovered from different portions of the oviduct. The germinative area, about 40 μm diameter, is localized in the region of the whitish polar spot, about 450 μm diameter. The structural changes in the germinative area are connected with the formation and extrusion of the first and second polar bodies. Of the two types of oviductal eggs observed, those covered with microvilli (type 1) were found only in the ostial portions of the oviduct, whilst those covered with microfolds (type 11) were found more distally. The structural difference between these two types may be related to the known reduction in surface area of the cell membrane during oocyte maturation.

Chen, YiPing; Solursh, Michael

doi: 10.1007/BF00190236pmid: 28306021

The time of determination of cartilage and skeletal muscle was studied by making chimeric grafts or explants of small tissue pieces from several stages of early chick or quail embryos. Chondrogenesis was assessed by histology or with antibodies directed against type II collagen or cartilage proteoglycan, while myogenesis was detected immunohistochemically with antibodies directed against 3 different muscle markers, including muscle myosin. Grafts from Hensen's node, primitive streak and segmental plate of donor embryos of Stage 3–5 (Hamburger and Hamilton) were transplanted under the ectoderm in the extraembryonic area of Stage 12 host embryos. In addition, explants and mesodermal cells were cultured on glass in DMEM+F12 medium supplemented with 10% FCS. The results showed that determined myogenic cells could first be detected in Hensen's node and the primitive streak at Stage 3+−4 and that they developed from mesodermal cells located between the epiblast and hypoblast. Myogenic cells also appeared in grafted and explanted segmental plate with or without notochord from Stage 5 embryos. On the other hand, cartilage cells only formed in grafted and explanted segmental plate that also contained notochord. RA (1 ng/ml) could induce the formation of cartilage cells in the explanted primitive streak without Hensen's node or notochord taken from Stage 3–5 embryos and could also promote the differentiation of myogenic cells in primitive streak from Stage 3 embryo. Thus RA can substitute for Hensen's node or the notochord in the induction of cartilage cells and has some stimulatory effects on the differentiation of myogenic cells. Additional evidence indicates that the hypoblast might play an “inductive” role in the formation of the notochord which may subsequently promote the differentiation of cartilage cells.

Granadino, Begoña; San Juán, Ana; Sánchez, Lucas

doi: 10.1007/BF00190237pmid: 28306022

In Drosophila melanogaster, the gene Sex-lethal (Sxl) controls the processes of sex determination, dosage compensation, oogenesis and sexual behaviour. The control of Sxl is by alternative splicing of its primary RNA. We have identified a gene, female-lethal-2-d (fl(2)d), which is needed for the female-specific splicing of Sxl RNA and which also has a vital function independent of Sxl. Here we analyse other aspects of the gene fl(2)d. Specifically, we have analysed the effect of the temperature-sensitive mutation fl(2)d 1 on the viability of adult flies homozygous for this mutation. We have found that the viability of the mutant females is reduced, while that of the mutant males is not affected. In addition, the capacity of the mutant females to be inseminated is considerably reduced, whilst all the mutant males are able to inseminate females. These effects on females are suppressed by Sxl M1. However, the fat body cells of fl(2)d 1 homozygous females are able to synthesize yolk proteins at the restrictive temperature. We have also carried out, in males, a clonal analysis of fl(2)d 2, a mutation lethal in both sexes. We have found that the clones are fully viable. We conclude that the gene fl(2)d seems to be necessary during the adult life of females for the processes that require Sxl + activity. Moreover, the Sxl-independent vital function of fl(2)d seems to be required in both sexes only during larval development.