The pattern of early cleavage of the marsupial frog Gastrotheca riobambae

Comparison of early development of the marsupial frog Gastrotheca riobambae with Xenopus laevis suggests that the cleavage pattern of Xenopus and the tight coupling of events at the midblastula transition are features of the accelerated development of small amphibian eggs with aquatic reproduction, rather than generalized features of amphibian development. The large eggs of the marsupial frog Gastrotheca riobambae not only display an atypical holoblastic pattern of cleavage and a very slow rate of development, but the events of the midblastula transition are uncoupled, suggesting that amphibians may have a diversity of developmental patterns. Early cleavage of the egg, which measures about 3 mm in diameter, occurs mostly by meridional and vertical furrows. The first cleavage cycle takes about 16 to 20 h and about four days may be required to reach the midblastula stage. Cleavage becomes asynchronous at about the third cleavage cycle evidenced by the formation of cleavage furrows. However, during cleavage (up to 342-cells), the majority of the nuclei divide synchronously and only 15 to 40% of the nuclei of a given embryo have a different cleavage schedule. At the 8-cell stage, nucleoli become visible (approximately 24 h after amplexus), signaling that transcription of rRNA has started at this early stage. Cell motility was detected in three- to four-day old embryos and seems to be associated with changes in cell shape and with expansion of the blastocoel at this stage.

Patterns of engrailed protein in early Drosophila embryos

By the onset of gastrulation during nuclear cycle 14 of Drosophila embryogenesis, the engrailed gene is expressed in fourteen one-cell-wide stripes. Each stripe defines the anlagen of the posterior compartment of a metameric segment. We report here several observations relating to the role and disposition of the engrailed protein during the embryonic stages that precede cellularization. We demonstrate that in embryos mutant for the engrailed gene, there were characteristic morphological abnormalities as early as the 6th cleavage cycle. In addition, the engrailed protein was detected in pre-cycle-9 embryos by Western blot analysis. When localization of engrailed protein begins during cycle 14, engrailed expression was first present in broad anterior and posterior regions before the fourteen-stripe pattern appeared.

Transition from mitosis to interphase in sea urchin first division: immunofluorescence studies of tubulin distribution in methacrylate sections.

Previous immunofluorescence studies of microtubule distribution in fertilized sea urchin eggs have suffered from poor resolution caused by cell thickness, unavoidable artifacts resulting from excessive flattening, or extraction by detergents of membranes and other lipid-containing structures that may be of interest in relation to the microtubules. To avoid these difficulties, we have developed a fixation and embedding protocol based on buffered paraformaldehyde fixation and butyl-methyl methacrylate embedment, which allows immunofluorescence staining of 0.5-1 micron sections. Polymerization artifacts are reduced by polymerizing the methacrylate at a relatively low temperature (40-45 degrees C) and by flat embedding for more uniform polymerization. Using this method, we have examined mitotic stages in the first cleavage cycle of the sea urchin Strongylocentrotus purpuratus. We provide evidence that the interphase microtubules that appear after first division are not derived from the mitotic asters but are new structures growing from organizing centers within the degenerating mitotic asters. During the transition from mitosis to interphase, there is a temporary overlap of old and new microtubules to form a very large composite aster at telophase before the old structure finally disappears.