Development Genes and Evolution

Shi, De -Li; Riou, Jean -François; Darribère, Thierry; Boucaut, Jean -Claude

doi: 10.1007/bf00380018pmid: 28305784

SummaryA polyclonal antibody (SP-2) has been produced, which recognizes antigens expressed in epidermal cells of Pleurodeles waltlii embryos. The antigens appear first at the end of gastrulation in the external surface of the embryo and are selectively expressed in ectodermally derived epidermal structures. Ectodermal commitment was investigated using cell cultures and blastocoel graft experiments. The four animal blastomeres of the 8-cell stage as well as the animal cap explants of the early gastrula stage cultured in vitro differentiate into epidermis, and SP-2 antigens are expressed. The expression of SP-2-defined antigens is inhibited both in vivo and in vitro by the inductive interaction of chordomesoderm. Once dissociated, ectodermal cells do not react with SP-2. Conversely, the aggregation of ectodermal cells may restore the expression of SP-2 antigens. Transplantation of animal cap explants or isolated ectodermal cells into the blastocoel of a host embryo at the early gastrula stage shows that only cells integrated into the epidermis express the marker antigens. When vegetal cells were dissociated from donor embryos before the mid-blastula stage and implanted into the blastocoel of host embryos at the early gastrula stage, their progeny were found in all germ layers, cells that were found in the host epidermis were stained with SP-2, whereas those contributing to mesoderm and endoderm were not. Thus the acquisition of cell polarity in epidermal differentiation and the organization of cells into epithelial structures are essential for SP-2-defined antigen expression.

Bellen, Hugo Jozef; Kiger, John Andrew

doi: 10.1007/bf00380019pmid: 28305785

SummaryThe developmental patterns of embryos produced by female germ line cells homozygous for null-enzyme mutations of dunce and for dunce in combination with each of three different rutabaga mutations are compared with the normal pattern. At least three discrete developmental defects at progressive stages following fertilization can be identified and correlated with the loss of adenylate cyclase activity caused by rutabaga mutations, suggesting that the defects are caused by elevated cyclic AMP levels in female germ line cells. The earliest defect occurs soon after fertilization and affects DNA replication and mitosis, prevents nuclear migration, and leads to large polyploid nuclei. A later defect prevents cleavage nuclei from migrating into, or dividing in, the posterior region of the egg. The last affects the developmental behavior or fate of blastoderm cells. Some of these defects mimic those produced by previously described maternal-effect mutations.

Fleig, Richard; Walldorf, Uwe; Gehring, Walter Jakob; Sander, Klaus

doi: 10.1007/bf00380020pmid: 28305786

SummaryWe have isolated and characterized a homeoboxcontaining gene from the honeybee Apis mellifera. Its homeobox region shows a high degree of sequence similarity to the homeobox of the Drosophila gene Deformed (Dfd). At the DNA level 82% of the basepairs are the same, whereas the putative amino acid sequences are identical between the bee and the fruitfly genes. Similarity is also present 5′ and 3′ to the homeobox. Using this isolate as a probe we have performed in situ hybridization on sections from blastoderm-stage embryos of the honeybee Apis mellifera. In early blastoderm stages we found a rather irregular pattern of labelled nuclei. In middle stages we found silver grains over each nucleus and also over the cytoplasm in a belt of blastoderm cells in the prospective gnathal region. These results indicate that the Deformed genes from honeybee and fruitfly are homologous both with respect to their DNA sequence and their spatial and temporal pattern of expression during embryogenesis.

Klein, Steven L.; King, Mary Lou

doi: 10.1007/bf00380021pmid: 28305787

SummaryThe proteins synthesized before the 512-cell stage by Xenopus blastomeres with different fates were compared by one dimensional PAGE. Blastomeres that contributed more progeny to antero-dorsal axial structures produced proportionately more of two proteins of 225000 and 245000 daltons. Additionally, these proteins were reversibly increased in ventralized embryos and were decreased in dorsalized embryos. These observations indicate that some proteins that are synthesized during cleavage stages are expressed to different degrees in different regions of the embryo, that their expression can be correlated to cell fate in the normal embryo, and that their expression is altered quantitatively in dorsalized and ventralized embryos. The inverse relationship between the production of these proteins and the potential to produce dorsal structures in the normal and in dorsalized/ventralized embryos is consistent with a model in which cell fate is influenced by a gradient of particular proteins.

Schierenberg, Einhard

doi: 10.1007/bf00380022pmid: 28305788

SummaryEarly embryogenesis of the nematode Caenorhabditis elegans is characterized by the continuous visibility of a germline and the stepwise separation of all somatic cells from it. Germline and somatic cells exhibit different cleavage patterns. Typical for the germline is a series of stemcell-like, unequal cleavages generating blastomeres, which differ in size, cell cycle periods, and fate. Typical for members of somatic cell lineages during early development are their equal and synchronous cleavages generating cells of similar appearance. Using a laser microbeam various experiments have been carried out to investigate the conditions that lead to the two different types of cleavage. Development of partial embryos demonstrates that the potential for germline-like cleavage is localized in the posterior region of the fertilized egg prior to both the formation of pronuclei and the posterior aggregation of germline-specific granules. Experimental alteration of the cleavage plane can result in a switch from unequal to equal cleavage, with an apparent correlation between the orientation of the mitotic spindle and the type of cleavage. Nuclear transfer experiments indicate that nuclei and centrioles are not involved in the decision as to which type of cleavage will be executed. Cytoplasmic transfer from soma-like to germline-like cleaving cells and vice versa does not alter the cleavage type in the recipient cell. But if separation of germline from soma is delayed after the removal of a centrosome, germline-like cleavage may be completely suppressed, all cells thereafter dividing soma-like.

Miner, Cristina; Represa, J. J.; Barbosa, E.; Giraldez, F.

doi: 10.1007/bf00380023pmid: 28305789

SummaryThe role of protein kinase C (PKC). during the early development of the inner ear was investigated using organ culture techniques. Otocysts isolated from chick embryos were made quiescent by culturing in the absence of serum for 24 h. The normal process of development could be reactivated by restoration of serum and other growth factors. Addition of phorbol ester (TPA) or synthetic diacylglycerol (OAG) to serum-free medium was also effective in reactivating development and stimulation of DNA synthesis was 41% and 52% of that of serum, respectively. Insulin potentiated the effects of TPA and OAG but had no effect when present alone. Morphogenesis and the associated cell proliferation stimulated by either serum or PKC activation were both inhibited by sphingosine, an in vitro inhibitor of PKC. Inhibition by sphingosine was dose-dependent with a half-maximal inhibitory concentration of about 10 μM. The results suggest that PKC activation is an essential step in controlling proliferative growth during early stages of the development of the inner ear.

Zagris, Nikolas; Matthopoulos, Demetrios

doi: 10.1007/bf00380024pmid: 28305790

SummaryThe polypeptides synthesized during the morula stage in the chick embryo are insensitive to transcriptional inhibition by α-amanitin. Protein synthesis seems to depend predominantely, if not exclusively, on the recruitment of maternal mRNA, rather than on embryonic gene expression in chick morula. The morula embryo expresses the heatshock polypeptides when stressed at 43°C. The heat-induced polypeptides are isoforms of polypeptides that are synthesized normally. These polypeptides are α-amanitin sensitive and appear to mark the first major expression of the embryonic genome in the chick embryo.

Grinfeld, Simone; Jacquet, Paul; Gilles, Jan; Baugnet-Mahieu, Lucile

doi: 10.1007/bf00380025pmid: 28305791

SummaryIn some strains of mice, eggs when X irradiated during the pronuclear stage, undergo a mitotic block in the G2 phase of the first cell cycle and cleave when the second division takes place in controls. The importance of this effect varies considerably with the strain and depends exclusively on the maternal genotype. In previous work, two-dimensional electrophoresis showed that eggs blocked at the one-cell stage after irradiation, undergo the same modifications in polypeptide synthesis as two-cell controls of the same age, except at the time of normal first mitosis, where three polypeptide sets of 30, 35 and 45 kDa appear only in cleaving controls. In the present study, we have found phosphorylations in dividing controls, on polypeptides of 30, 35 and 45 kDa. These phosphorylations are not seen in blocked irradiated eggs.