Binding of anti-fibronectin to early amphibian ectoderm does not result in inhibition of neural induction under in vitro conditionsGrunz, Horst; Darribère, Thierry; Boucaut, Jean
doi: 10.1007/BF00376344pmid: 28305695
Antibodies directed to fibronectin (anti-FN) were injected into the blastocoel of late blastulae of Xenopus laevis. Two animal caps (ectoderm) were isolated, when control embryos reached the early gastrula stage, and were combined with untreated upper blastopore lip in the sandwich method. In two control series fibronectin or Holtfreter solution was injected into the blastocoel. The results of the experiments suggest that neural induction cannot be prevented by binding anti-FN to fibronectin, which covers the blastocoelic side of the ectoderm. The data support the view that extracellular matrix proteins are not themselves responsible for neural induction. However, in comparison with the control series a slight shift of the differentiation pattern in the spinocaudal direction could be observed in the anti-FN series. The possible role of extracellular proteins in the formation of a close juxtaposition of mesodermal and ectodermal target cells as a prerequisite for shortdistance transmission of neural inducers is discussed.
Fertile heteroallelic combinations of mutant alleles of the otu locus of Drosophila melanogasterStorto, Patrick; King, Robert
doi: 10.1007/BF00376345pmid: 28305696
This paper describes the ovarian pathologies observed when 108 different heteroallelic combinations were made involving 17 independent mutations at the ovarian tumor (otu) locus. Most of the mutant phenotypes can be explained as graded responses by individual germ cells to different levels of functionally active otu gene product (OGP) synthesized by the mutant cells themselves. The lowest and highest levels of OGP appear to be produced by otu
10 and otu
14, respectively. In most heteroallelic ovaries the alleles have additive effects, and hybrid germ cells reach a developmental stage more advanced than the “weaker” homozygote but less advanced than the “stronger” homozygote. However, examples of both positive and negative complementation also have been found, and these suggest that the products encoded by different mutant alleles can combine to form dimers or multimers which may be superior or inferior to the homodimers. In flies homozygous for otu
11 most ovarioles contain tumors, but some germ cells are able to develop further than those in otu
14 homozygotes. This suggests that, while otu
11 produces intermediate levels of OGP, it also produces a second product (which otu
14 cannot make) that is utilized at the period in oogenesis when development in cells homozygous for otu
14 is blocked. When otu
11 is combined with any one of eight specific alleles, it allows oocyte/nurse cell syncytia to differentiate that can complete development and undergo embryogenesis, if fertilized. The endopolyploid nurse cells of these hybrids have giant polytene chromosomes, and the presence of GPCs in functionally active, germ-line derived cells provides an interesting new system for experimental study. Analysis of the characteristic ovarian pathologies produced by flies of different genotypes leads to the conclusion that the products of the otu
+ gene are utilized during at least six different periods in Drosophila oogenesis.
Lineage analysis of transplanted individual cells in embryos of Drosophila melanogasterBeer, Justinus; Technau, Gerhard; Campos -Ortega, Jose
doi: 10.1007/BF00376346pmid: 28305697
We describe the results of cell transplantation experiments performed to investigate mesodermal lineages in Drosophila melanogaster, particularly the lineages of the somatic muscles, the visceral muscles and the fat body. Cells to be transplanted were labelled by injecting a mixture of horseradish peroxidase (HRP) and fluorescein-dextran (FITC) in wild-type embryos at the syncytial blastoderm stage. For transplantation cells were removed from the ventral furrow, 8–12 min after the start of gastrulation, and individually transplanted into homotopic or heterotopic locations of unlabelled wild-type hosts of the same age. HRP labelling in the resulting cell clones was demonstrated histochemically in the fully developed embryo; histotypes could be distinguished without ambiguity. Mesodermal cells were already found to be committed to mesodermal fates at the time of transplantation. They developed only into mesodermal derivatives and did not integrate in non-mesodermal organs upon heterotopical transplantation. No evidence was found for commitment to any particular mesodermal organ at the time of transplantation. The majority of somatic muscle clones contributed cells to only one segment. However, clones were not infrequently distributed through two or even three segments. Clones of fat body cells were generally restricted to a small region. However, cells of clones of visceral musculature were widely distributed. With respect to the proliferative abilities of transplanted cells the clones were difficult to interpret due to the syncytial character of the somatic musculature and the fact that the organization of the other organs is poorly understood. Evidence from histological observations of developing normal embryos indicates only three mitoses for mesodermal cells. Clones larger than seven cells were not found when embryos were fixed previous to germ-band shortening; larger clones were found in the fat body and visceral musculature after fixing the embryos at the end of organogenesis. Quantitative considerations suggest that a few mesodermal cells might perform more than three mitoses.
Genetic studies of mutations at two loci of Drosophila melanogaster which cause a wide variety of homeotic transformationsShearn, Allen; Hersperger, Evelyn; Hersperger, Grafton
doi: 10.1007/BF00376347pmid: 28305698
The ash-1 locus is in the proximal region of the left arm of the third chromosome of Drosophila melanogaster and the ash-2 locus is in the distal region of the right arm of the third chromosome. Mutations at either locus can cause homeotic transformations of the antenna to leg, proboscis to leg and/or antenna, dorsal prothorax to wing, first and third leg to second leg, haltere to wing, and genitalia to leg and/or antenna. Mutations at the ash-1 locus cause, in addition, transformations of the posterior wing and second leg to anterior wing and second leg, respectively. A similar spectrum of transformations is caused by mutations at yet another third chromosome locus, trithorax. One extraordinary aspect of mutations at all three of these loci is that they cause such a wide variety of transformations. For mutations at both of the loci that we have studied the expression of the homeotic phenotype is both disc-autonomous (as shown by injecting mutant discs into metamorphosing larvae) and cell autonomous (as shown by somatic recombination analysis). The original mutations which identified these two loci, although lethal, manifest variable expressivity and incomplete penetrance of the homeotic phenotype suggesting that they are hypomorphic. The phenotype of double mutants which were synthesized by combining different pairs of those original mutations manifest for two of the four pairs a greater degree of expressivity and slightly more penetrance of the homeotic transformations. This mutual enhancement suggests that the products of both loci interact in the same process. A third double mutant expresses a discless phenotype.
Low-molecular-weight factors from colonial hydroids affect pattern formationPlickert, Günter
doi: 10.1007/BF00376349pmid: 28305700
Two morphogenetic factors have been isolated from tissue of colonial hydroids. Both exert strong effects on pattern formation during metamorphosis, regeneration and colony development. Polyp-inhibiting factor (PIF) is a bivalent inhibitor which strongly affects head and bud formation but acts weakly on stolon branching. Proportion-altering factor (PAF) is a distalizing factor. It counteracts the formation of stolon and promotes the formation of head structures during metamorphosis and regeneration. PIF and PAF antagonistically influence the spatial arrangement of polyps within a colony. They are capable of dislocating structures and thus appear to interfere with or are even part of the pattern-controlling mechanism. Both factors are of low molecular size (about 500 daltons), hydrophilic and probably not peptides.
Effects of thyroxine on the ontogeny of the vitellogenic response in Pleurodeles waltli of both sexesCayrol, Christian; Deparis, Pierre
doi: 10.1007/BF00376350pmid: 28305701
In the newt, Pleurodeles waltli, vitellogenin synthesis can be induced by oestrogen prior to metamorphosis and well before vitellogenin synthesis is normally observed in females. Vitellogenin indicubility has been shown to occur during normal development at stage 53 in both sexes. Immersion of young larvae in thyroxine at concentrations that did not result in early morphological metamorphosis led to a precocious acquisition of liver responsiveness to oestrogen. Treatment of larvae with thiourea greatly diminished the ability of oestradiol to induce vitellogenin synthesis, although it did not abolish the hormonal response.
Isolation of sea urchin embryo cell surface membranes on polycationic beadsHelmly, R.; Brown, Kenneth
doi: 10.1007/BF00376351pmid: 28305702
Blastula cell surface membranes of the sea urchin, Strongylocentrotus purpuratus, were isolated on polycationic beads by a method modified from Jacobson and Branton (1977) and Jacobson (1980). This study represents the first application of this procedure to an embryonic system. Embryo cells were attached to polylysine-coated polyacrylamide beads and lysed, leaving the embryo cell surface membranes still attached to the beads, and cytoplasmic particles were washed free of the exposed inner surfaces of the membranes. Cell surface membrane sheets were desorbed from the beads and collected by centrifugation. Approximately 8% and 5% of the cell surface membranes of dissociated embryo cells were recovered on the beads and in the membrane pellet, respectively. Specific activities of [3H]concanavalin A-binding and of the cell surface marker enzymes, alkaline phosphatase and Na+/K+ ATPase, were 16-, 19-, and 32-fold higher, respectively, in the cell surface membrane fraction than in the embryo cell homogenate. Membranes were relatively free of cytoplasmic contaminants as judged from electron micrographs and enzyme analysis. Activities in the membrane fraction of the cytoplasmic marker enzymes, cytochrome c oxidase, catalase, acid phosphatase, NADP- and NADPH-cytochrome c reductase, and acetylcholinesterase, were substantially less than homogenate levels. The entire procedure can be completed in 4 h. Since this cell surface membrane isolation technique relies only on the tendency of a negatively charged cell to adhere to a positively charged surface, it is less likely than most other methods to exhibit species and developmental stage specificity and should prove useful in the study of the developmental role of embryonic stage-specific membrane components.