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At least 163 records · Page 9
All sourcesNASA NTRSDOE OSTI
DOE OSTI2024

Organogenetic transcriptomes of the Drosophila embryo at single cell resolution

To gain insight into the transcription programs activated during the formation of Drosophila larval structures, we carried out single cell RNA sequencing during two periods of Drosophila embryogenesis: stages 10-12, when most organs are first specified and initiate morphological and physiological specialization; and stages 13-16, when organs achieve their final mature architectures and begin to function. Our data confirm previous findings with regards to functional specialization of some organs – the salivary gland and trachea – and clarify the embryonic functions of another – the plasmatocytes. We also identify two early developmental trajectories in germ cells and uncover a potential role for proteolysis during germline stem cell specialization. We identify the likely cell type of origin for key components of the Drosophila matrisome and several commonly used Drosophila embryonic cell culture lines. Finally, we compare our findings with other recent related studies and with other modalities for identifying tissue-specific gene expression patterns. These data provide a useful community resource for identifying many new players in tissue-specific morphogenesis and functional specialization of developing organs.

59 BASIC BIOLOGICAL SCIENCES↗
NASA NTRS1985

Effects on in Vivo and in Vitro Exposure to Excess Gravity on Growth and Differentiation of Mammalian Embryos

Studies on the development of embryonic mouse tissues exposed to excess gravity in vitro and in vivo are discussed. Suppression is seen in limb buds cultured under 3G. Mouse palates were exposed to excess G in vitro, 13- and 14-day palates were exposed to 2.6G for 24 hours. For in vivo studies, a small animal centrifuge was constructed. When the centrifuge is operated at 40 and 45 rpm, the linear accelerations generated range from 1.8 to 3.5G. The effects of gravity on body weights and on reproduction is also presented.

Duke, J.↗
NASA NTRS1989

Gravity and animal embryos

Out of more than 4,500 rat hours in space there was only one experimental attempt (Cosmos 1129) at mating with an apparent absence of fertilization, implantation and subsequent development to term and partuition. Portions of this process were successfully flown, however, including the major portion of organogenesis in the rat (Cosmos 1524). These observations show that the cellular and molecular events underlying morphogenesis and differentiation in a small mammal can proceed normally in-utero under microgravity and other conditions encountered during short-duration flight. However, it is not known whether this situation will hold for larger mammals over several generations during extended missions that venture outside of near Earth. Furthermore, it is not understood why the previous attempt at obtaining copulation, fertilization and implantation in orbit failed but may be related to limitations of the rat habitat for meeting the preconditions for reproductive behavior. With respect to mammalian development it is important to appreciate that fertilization and development occur internally within the female and take a long time to complete and their success will, therefore, be contingent upon the maternal response to the space environment. One process central to development (the establishment of cell lines) is initiated prior to implantation by environmental asymmetries preceived by progenitor cells. These asymmetries appear to result from the formation of asymmetric cell-cell contacts and the concommitant development of an electrical axis across the progenitor cells. Other asymmetries were also documented. It is not known whether any of the known asymmetries perceived by progenitor cells are influenced by gravity vectors and/or by the maternal response to microgravity and other conditions encountered in space.

Wiley, Lynn M.↗
NASA NTRS1992

Eggs: The role of gravity in the establishment of the dorso-ventral axis in the amphibian embryo (7-IML-1)

The purpose of this experiment is to fertilize frog (Xenopus laevis) eggs under microgravity, to perform histological fixations at two different programmed times, and after return to Earth, to determine whether timing and pattern of egg cleavages and axis formation are normal. Because of the limited viability of the gametes, this experiment will be the very first to be activated in the Biorack. Various aspects of this investigation are discussed.

Ubbels, Geertje A.↗
NASA NTRS1994

Early development of Xenopus embryos is affected by simulated gravity

Early amphibian (Xenopus laevis) development under clinostat-simulated weightlessness and centrifuge-simulated hypergravity was studied. The results revealed significant effects on (i) 'morphological patterning' such as the cleavage furrow pattern in the vegetal hemisphere at the eight-cell stage and the shape of the dorsal lip in early gastrulae and (ii) 'the timing of embryonic events' such as the third cleavage furrow completion and the dorsal lip appearance. Substantial variations in sensitivity to simulated force fields were observed, which should be considered in interpreting spaceflight data.

Yokota, Hiroki↗
NASA NTRS1995

Modification of Experimental Protocols for a Space Shuttle Flight and Applications for the Analysis of Cytoskeletal Structures During Fertilization, Cell Division , and Development in Sea Urchin Embryos

To explore the role of microgravity on cytoskeletal organization and skeletal calcium deposition during fertilization, cell division, and early development, the sea urchin was chosen as a model developmental system. Methods were developed to employ light, immunofluorescence, and electron microscopy on cultures being prepared for flight on the Space Shuttle. For analysis of microfilaments, microtubules, centrosomes, and calcium-requiring events, our standard laboratory protocols had to be modified substantially for experimentation on the Space Shuttle. All manipulations were carried out in a closed culture chamber containing 35 ml artificial sea water as a culture fluid. Unfertilized eggs stored for 24 hours in these chambers were fertilized with sperm diluted in sea water and fixed with concentrated fixatives for final fixation in formaldehyde, taxol, EGTA, and MgCl2(exp -6)H2O for 1 cell to 16 cell stages to preserve cytoskeletal structures for simultaneous analysis with light, immunofluorescence, and electron microscopy, and 1.5 percent glutaraldehyde and 0.4 percent formaldehyde for blastula and plueus stages. The fixed samples wre maintained in chambers without degradation for up to two weeks after which the specimens were processed and analyzed with routine methods. Since complex manipulations are not possible in the closed chambers, the fertilization coat was removed from fixation using 0.5 percent freshly prepared sodium thioglycolate solution at pH 10.0 which provided reliable immunofluorescence staining for microtubules. Sperm/egg fusion, mitosis, cytokinesis, and calcium deposition during spicule formatin in early embryogenesis were found to be without artificial alterations when compared to cells fixed fresh and processed with conventional methods.

Chakrabarti, Amitabha↗
NASA NTRS2002

Dynamical Evolution of Planetary Embryos

During the past decade, progress has been made by relating the 'standard model' for the formation of planetary systems to computational and observational advances. A significant contribution to this has been provided by this grant. The consequence of this is that the rigor of the physical modeling has improved considerably. This has identified discrepancies between the predictions of the standard model and recent observations of extrasolar planets. In some cases, the discrepancies can be resolved by recognition of the stochastic nature of the planetary formation process, leading to variations in the final state of a planetary system. In other cases, it seems more likely that there are major deficiencies in the standard model, requiring our identifying variations to the model that are not so strongly constrained to our Solar System.

Wetherill, George W.↗
NASA NTRS1998

Plants and somatic embryos in space: what have we learned?

Space provides a unique environment that can affect the interplay between cell cycle controls and environment and can thus modify the processes of cell division, development and growth. It is proposed that the chromosomal and nuclear abnormalities frequently encountered in cells of various plants exposed to space are due to a combination of factors including the biological status of the systems and the way in which they are grown, exposed to, and ultimately, the way in which they experience multiple stresses. The extent to which space-specific changes become manifest is dependent on the extent of pre-existing stresses in the system. This has become evident in a variety of plant species grown in space but has been particularly amenable to study using in vitro systems, especially in developing embryoids. The following observations allow us to harmonize disparate results from a variety of space experiments:- (a) the more completely developed a system, the less likely it is to show cell stress during growth; the less morphologically complex, the greater the vulnerability; (b) the size/"packaging" of the genome (karyotype) are significant experimental variables; plants with larger genomes (e.g. polyploids) seem to be more space-stress tolerant; (c) a single space-associated stress is inadequate to produce a significant adverse response unless the stress is severe, or a biological parameter necessary to 'amplify' it exists. On this view, an appropriate "stress match" with other non-equilibrium determinants, much like a 'tug of war', can result in genomic variations in space. All this emphasizes that fastidiously-controlled growing environments must be devised if one is to resolve the matter of direct versus indirect effects of space. Better understanding of the novel physico-chemical equilibrium phenomena associated with space will allow those interested in space cell and developmental biology to pick and choose procedures best suited to their exploitation for specific objectives.

Flight Experiment↗